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1

Inhibitory mechanism of chroman compound on LPS-induced nitric oxide production and nuclear factor-?B activation  

Microsoft Academic Search

6-Hydroxy-7-methoxychroman-2-carboxylic acid phenylamide (KL-1156) is a novel chemically synthetic compound. In the present study, the chroman KL-1156 compound was found to inhibit lipopolysaccharide (LPS)-induced nitric oxide production in macrophages RAW 264.7. KL-1156 compound attenuated LPS-induced synthesis of both mRNA and protein of inducible nitric oxide synthase (iNOS), in parallel, and inhibited LPS-induced iNOS promoter activity, indicating that the chroman compound

Byung Hak Kim; Alavala Matta Reddy; Kum-Ho Lee; Eun Yong Chung; Sung Min Cho; Heesoon Lee; Kyung Rak Min; Youngsoo Kim

2004-01-01

2

Inhibition of Nitric Oxide Production Rescues LPS-Induced Fetal Abortion in Mice  

Microsoft Academic Search

In this report, we examined the involvement of the cytokines tumor necrosis factor (TNF)-?, interferon (IFN)-?, interleukin (IL)-4, and IL-10 as well as nitric oxide (NO) in the lipopolysaccharide (LPS)-induced experimental abortion model in BALB\\/c mice. Although in vivo administration of LPS in pregnant mice showed a 72% decrease of serum IL-10, no significant difference in serum TNF-?, IFN-?, and

I. Athanassakis; I. Aifantis; A. Ranella; K. Giouremou; S. Vassiliadis

1999-01-01

3

Folimycin (concanamycin A) inhibits LPS-induced nitric oxide production and reduces surface localization of TLR4 in murine macrophages  

Microsoft Academic Search

Lipopolysaccharide (LPS) is a major cell wall component of Gram-negative bacteria and signals through a receptor complex which consists of TLR4, MD-2 and CD14. LPS signaling in macrophages induces the production of many pro-inflammatory molecules, including nitric oxide (NO). In this study, we have shown that folimycin, a macrolide antibiotic and a specific inhibitor of vacuolar ATPase (V-ATPase), inhibits LPS-induced

Sandeepa M. Eswarappa; Nirmalya Basu; Omana Joy; Dipshikha Chakravortty

2008-01-01

4

Suppression of LPS-induced Interferon-? and nitric oxide in splenic lymphocytes by select estrogen-regulated microRNAs: a novel mechanism of immune modulation  

PubMed Central

MicroRNAs (miRNAs), recently identified noncoding small RNAs, are emerging as key regulators in homeostasis of the immune system. Therefore, aberrant expression of miRNAs may be linked to immune dysfunction, such as in chronic inflammation and autoimmunity. In this study, we investigated the potential role of miRNAs in estrogen-mediated regulation of innate immune responses, as indicated by up-regulation of lipopolysaccharide (LPS)–induced interferon-gamma (IFN?), inducible nitric oxide synthase (iNOS), and nitric oxide in splenic lymphocytes from estrogen-treated mice. We found that miR-146a, a negative regulator of Toll-like receptor (TLR) signaling, was decreased in freshly isolated splenic lymphocytes from estrogen-treated mice compared with placebo controls. Increasing the activity of miR-146a significantly inhibited LPS-induced IFN? and iNOS expression in mouse splenic lymphocytes. Further, miRNA microarray and real-time reverse transcriptase–polymerase chain reaction (RT-PCR) analysis revealed that estrogen selectively up-regulates/down-regulates the expression of miRNAs in mouse splenic lymphocytes. miR-223, which is markedly enhanced by estrogen, regulates LPS-induced IFN?, but not iNOS or nitric oxide in splenic lymphocytes. Inhibition of miR-223 activity decreased LPS-induced IFN? in splenic lymphocytes from estrogen-treated mice. Our data are the first to demonstrate the selective regulation of miRNA expression in immune cells by estrogen and are indicative of an important role of miRNAs in estrogen-mediated immune regulation. PMID:18791161

Dai, Rujuan; Phillips, Rebecca A.; Zhang, Yan; Khan, Deena; Crasta, Oswald

2008-01-01

5

Hypericum triquetrifolium—Derived Factors Downregulate the Production Levels of LPS-Induced Nitric Oxide and Tumor Necrosis Factor-? in THP-1 Cells  

PubMed Central

Based on knowledge from traditional Arab herbal medicine, this in vitro study aims to examine the anti-inflammatory mechanism of Hypericum triquetrifolium by measuring the expression and release of pro-inflammatory cytokines, tumor necrosis factor-? (TNF-?) and interleukine-6 (IL-6), and inducible nitric oxide synthase (iNOS) in human monocytic cells, THP-1. The effects were assessed by measuring the levels of secretory proteins and mRNA of TNF-? and IL-6, the levels of nitric oxide (NO) secretion and the expression of iNOS in THP-1 cells. Cells were treated with 5??g lipopolysaccharide/ml (LPS) in the presence and absence of increasing concentrations of extracts from the aerial parts of H. triquetrifolium. During the entire experimental period, we used extract concentrations (up to 250??g?mL?1) that had no cytotoxic effects, as measured with MTT and LDH assays. Hypericum triquetrifolium extracts remarkably suppressed the LPS-induced NO release, significantly attenuated the LPS-induced transcription of iNOS and inhibited in a dose-dependent manner the expression and release of TNF-?. No significant effects were observed on the release of IL-6. Taken together, these results suggest that H. triquetrifolium probably exerts anti-inflammatory effects through the suppression of TNF-? and iNOS expressions. PMID:18955363

Saad, Bashar; AbouAtta, Bernadette Soudah; Basha, Walid; Hmade, Alaa; Kmail, Abdalsalam; Khasib, Said; Said, Omar

2011-01-01

6

Pheophytin a Inhibits Inflammation via Suppression of LPS-Induced Nitric Oxide Synthase-2, Prostaglandin E2, and Interleukin-1? of Macrophages  

PubMed Central

Inflammation is a serious health issue worldwide that induces many diseases such as sepsis. There has been a vast search for potentially effective drugs to decrease mortality from sepsis. Pheophytin a is a chlorophyll-related compound derived from green tea. We found that pre-treatment with pheophytin a suppressed lipopolysaccharide (LPS)-induced nitric oxide (NO), prostaglandin E2 (PGE2), and interleukin-1? in RAW 264.7 macrophages. NO synthase-2 (NOS2) and cyclooxygenase-2 (COX-2) expression levels were repressed by pre-treatment with pheophytin a at both the transcriptional and translational levels. Pheophytin a inhibited NOS2 promoter activity, but not its mRNA stability, through extracellular signal-regulated kinase (ERK1/2). This suppression was reversed by ERK1/2 inhibitor (U0126). Pheophytin a reduced signal transducers and activators of transcription 1 (STAT-1) activation, without an obvious influence on activator protein-1 (AP-1) and nuclear factor ?B (NF-?B). These results suggest that pheophytin a functions by down-regulating the transcriptional levels of inflammatory mediators and blocking the ERK and STAT-1 pathways. PMID:25501336

Lin, Chun-Yu; Lee, Chien-Hsing; Chang, Yu-Wei; Wang, Hui-Min; Chen, Chung-Yi; Chen, Yen-Hsu

2014-01-01

7

Propofol attenuates LPS-induced tumor necrosis factor-?, interleukin-6 and nitric oxide expression in canine peripheral blood mononuclear cells possibly through down-regulation of nuclear factor (NF)-?B activation  

PubMed Central

Sepsis is a major cause of mortality in intensive care medicine. Propofol, an intravenous general anesthetic, has been suggested to have anti-inflammatory properties and able to prevent sepsis induced by Gram-positive and Gram-negative bacteria by down-regulating the gene expression of pro-inflammatory cytokines. However, propofol’s anti-inflammatory effects upon canine peripheral blood mononuclear cells (PBMCs) have not yet been clarified. Here, we isolate canine PBMCs and investigate the effects of propofol on the gene expressions of both lipopolysaccharide (LPS)-induced interleukin-6 (IL-6) and tumor necrosis factor (TNF)-? and upon the production of nitric oxide (NO). Through real-time quantitative PCR and the Griess reagent system, we found that non-cytotoxic levels of propofol significantly inhibited the release of NO and IL-6 and TNF-? gene expression in LPS-induced canine PBMCs. Western blotting revealed that LPS does significantly increase the expression of inducible NO synthase (iNOS) protein in canine PBMCs, while pretreatment with propofol significantly decreases the LPS-induced iNOS protein expression. Propofol, at concentration of 25 µM and 50 µM, also significantly inhibited the LPS-induced nuclear translocation of nuclear factor (NF)-?B p65 protein in canine PBMCs. This diminished TNF-?, IL-6 and iNOS expression, and NO production was in parallel to the respective decreased NF-?B p65 protein nuclear translocation in the LPS-activated canine PBMCs pretreated with 25 µM and 50 µM propofol. This suggests that non-cytotoxic levels of propofol pretreatment can down-regulate LPS-induced inflammatory responses in canine PBMCs, possibly by inhibiting the nuclear translocation of the NF-?B p65 protein. PMID:25312048

PEI, Zengyang; WANG, Jinqiu

2014-01-01

8

Synthesis of aurones and their inhibitory effects on nitric oxide and PGE2 productions in LPS-induced RAW 264.7 cells.  

PubMed

Sulfuretin is one of major constituents of Rhus verniciflua that exerts anti-inflammatory activities. Some of aurones were synthesized as sulfuretin derivatives and evaluated for their abilities to inhibit NO and PGE(2) production in LPS-induced RAW 264.7 cells in order to reveal the relationship. Of the aurones synthesized in the present study, 2h and 2i, which possess C-6 hydroxyl group in A-ring and methoxy substituents in B-ring, more potently inhibited NO and PGE(2) production and were less cytotoxic than sulfuretin. PMID:21723122

Shin, Seo Young; Shin, Min Cheol; Shin, Ji-Sun; Lee, Kyung-Tae; Lee, Yong Sup

2011-08-01

9

LPS-induced immune response in Drosophila  

Microsoft Academic Search

The study of the regulation of the inducible synthesis of antimicrobial peptides in Drosophila melanogaster has established this insect as a powerful model in which to study innate immunity. In particular, the molecular characterization of the regulatory pathway controlling the antifungal peptide drosomycin has revealed the importance of Toll receptors in innate immunity. We report here that injection of LPS

Jean-Luc Imler; Servane Tauszig; Emmanuelle Jouanguy; Claire Forestier; Jules A. Hoffmann

2000-01-01

10

Achillea millefolium L. Essential Oil Inhibits LPS-Induced Oxidative Stress and Nitric Oxide Production in RAW 264.7 Macrophages  

PubMed Central

Achillea millefolium L. is a member of the Asteraceae family and has been used in folk medicine in many countries. In this study, 19 compounds in A. millefolium essential oil (AM-EO) have been identified; the major components are artemisia ketone (14.92%), camphor (11.64%), linalyl acetate (11.51%) and 1,8-cineole (10.15%). AM-EO can suppress the inflammatory responses of lipopolysaccharides (LPS)-stimulated RAW 264.7 macrophages, including decreased levels of cellular nitric oxide (NO) and superoxide anion production, lipid peroxidation and glutathione (GSH) concentration. This antioxidant activity is not a result of increased superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) activities, but rather occurs as a result of the down-regulation of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor-? (TNF-?), interleukin-6 (IL-6) and heme oxygenase-1 (HO-1) expression, thus reducing the inflammatory response. Therefore, AM-EO can be utilized in many applications, including the treatment of inflammatory diseases in the future. PMID:23797659

Chou, Su-Tze; Peng, Hsin-Yi; Hsu, Jaw-Cherng; Lin, Chih-Chien; Shih, Ying

2013-01-01

11

Achillea millefolium L. Essential Oil Inhibits LPS-Induced Oxidative Stress and Nitric Oxide Production in RAW 264.7 Macrophages.  

PubMed

Achillea millefolium L. is a member of the Asteraceae family and has been used in folk medicine in many countries. In this study, 19 compounds in A. millefolium essential oil (AM-EO) have been identified; the major components are artemisia ketone (14.92%), camphor (11.64%), linalyl acetate (11.51%) and 1,8-cineole (10.15%). AM-EO can suppress the inflammatory responses of lipopolysaccharides (LPS)-stimulated RAW 264.7 macrophages, including decreased levels of cellular nitric oxide (NO) and superoxide anion production, lipid peroxidation and glutathione (GSH) concentration. This antioxidant activity is not a result of increased superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) activities, but rather occurs as a result of the down-regulation of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor-? (TNF-?), interleukin-6 (IL-6) and heme oxygenase-1 (HO-1) expression, thus reducing the inflammatory response. Therefore, AM-EO can be utilized in many applications, including the treatment of inflammatory diseases in the future. PMID:23797659

Chou, Su-Tze; Peng, Hsin-Yi; Hsu, Jaw-Cherng; Lin, Chih-Chien; Shih, Ying

2013-01-01

12

Sulfuretin isolated from heartwood of Rhus verniciflua inhibits LPS-induced inducible nitric oxide synthase, cyclooxygenase-2, and pro-inflammatory cytokines expression via the down-regulation of NF-kappaB in RAW 264.7 murine macrophage cells.  

PubMed

It has been reported that Rhusverniciflua exhibits anti-inflammatory, anti-oxidant and anti-cancer activities. However, little is known about biological activity of sulfuretin, a flavonoid isolated from R.verniciflua. In the present study, we investigated the anti-inflammatory effect and the underlying molecular mechanisms of sulfuretin in lipopolysaccharide (LPS)-induced RAW 264.7 cells. Sulfuretin dose-dependently reduced the productions of nitric oxide (NO), prostaglandin E(2) (PGE(2)), tumor necrosis factor-alpha (TNF-alpha), and interleukin-1 beta (IL-1 beta) induced by LPS. Consistent with these findings, sulfuretin significantly suppressed the LPS-induced expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), TNF-alpha, and IL-1 beta. In addition, sulfuretin attenuated LPS-induced DNA binding and the transcriptional activities of nuclear factor-kappa B (NF-kappaB), which was accompanied by a parallel reduction of degradation and phosphorylation of inhibitory kappa B-alpha (I kappaB-alpha) and consequently by decreased nuclear translocation of p65 subunit of NF-kappaB. Furthermore, pretreatment with sulfuretin significantly inhibited the LPS-stimulated activation of I kappaB kinase beta (IKK beta). Taken together, these results suggest that the anti-inflammatory effect of sulfuretin in LPS-treated RAW 264.7 macrophages is associated with the suppression of NF-kappaB transcriptional activity via the inhibitory regulation of IKKbeta phosphorylation. PMID:20546946

Shin, Ji-Sun; Park, Young Mi; Choi, Jung-Hye; Park, Hee-Juhn; Shin, Min Cheol; Lee, Yong Sup; Lee, Kyung-Tae

2010-08-01

13

Inhibition of IRAK-4 activity for rescuing endotoxin LPS-induced septic mortality in mice by lonicerae flos extract  

SciTech Connect

Highlights: •Lonicerae flos extract (HS-23) is a clinical candidate, Phase I for sepsis treatment. •Here, HS-23 or its major constituents rescued LPS-induced septic mortality in mice. •As a mechanism, they directly inhibited IRAK-4-catalyzed kinase activity. •Thus, they suppressed LPS-induced expression of NF-?B/AP-1-target inflammatory genes. -- Abstract: Lonicerae flos extract (HS-23) is a clinical candidate currently undergoing Phase I trial in lipopolysaccharide (LPS)-injected healthy human volunteers, but its molecular basis remains to be defined. Here, we investigated protective effects of HS-23 or its major constituents on Escherichia coli LPS-induced septic mortality in mice. Intravenous treatment with HS-23 rescued LPS-intoxicated C57BL/6J mice under septic conditions, and decreased the levels of cytokines such as tumor necrosis factor ? (TNF-?), interleukin (IL)-1? and high-mobility group box-1 (HMGB-1) in the blood. Chlorogenic acid (CGA) and its isomers were assigned as major constituents of HS-23 in the protection against endotoxemia. As a molecular mechanism, HS-23 or CGA isomers inhibited endotoxin LPS-induced autophosphorylation of the IL-1 receptor-associated kinase 4 (IRAK-4) in mouse peritoneal macrophages as well as the kinase activity of IRAK-4 in cell-free reactions. HS-23 consequently suppressed downstream pathways critical for LPS-induced activation of nuclear factor (NF)-?B or activating protein 1 (AP-1) in the peritoneal macrophages. HS-23 also inhibited various toll-like receptor agonists-induced nitric oxide (NO) production, and down-regulated LPS-induced expression of NF-?B/AP-1-target inflammatory genes in the cells. Taken together, HS-23 or CGA isomers exhibited anti-inflammatory therapy against LPS-induced septic mortality in mice, at least in part, mediated through the inhibition of IRAK-4.

Park, Sun Hong; Roh, Eunmiri [College of Pharmacy, Chungbuk National University, Cheongju 361-763 (Korea, Republic of)] [College of Pharmacy, Chungbuk National University, Cheongju 361-763 (Korea, Republic of); Kim, Hyun Soo [Pharmaceutical R and D Center, Huons Co., Ltd., Anyang (Korea, Republic of)] [Pharmaceutical R and D Center, Huons Co., Ltd., Anyang (Korea, Republic of); Baek, Seung-Il [College of Pharmacy, Chungbuk National University, Cheongju 361-763 (Korea, Republic of)] [College of Pharmacy, Chungbuk National University, Cheongju 361-763 (Korea, Republic of); Choi, Nam Song [Pharmaceutical R and D Center, Huons Co., Ltd., Anyang (Korea, Republic of)] [Pharmaceutical R and D Center, Huons Co., Ltd., Anyang (Korea, Republic of); Kim, Narae; Hwang, Bang Yeon; Han, Sang-Bae [College of Pharmacy, Chungbuk National University, Cheongju 361-763 (Korea, Republic of)] [College of Pharmacy, Chungbuk National University, Cheongju 361-763 (Korea, Republic of); Kim, Youngsoo, E-mail: youngsoo@chungbuk.ac.kr [College of Pharmacy, Chungbuk National University, Cheongju 361-763 (Korea, Republic of)] [College of Pharmacy, Chungbuk National University, Cheongju 361-763 (Korea, Republic of)

2013-12-13

14

The Protective Effect of Melatonin on Neural Stem Cell against LPS-Induced Inflammation  

PubMed Central

Stem cell therapy for tissue regeneration has several limitations in the fact that transplanted cells could not survive for a long time. For solving these limitations, many studies have focused on the antioxidants to increase survival rate of neural stem cells (NSCs). Melatonin, an antioxidant synthesized in the pineal gland, plays multiple roles in various physiological mechanisms. Melatonin exerts neuroprotective effects in the central nervous system. To determine the effect of melatonin on NSCs which is in LPS-induced inflammatory stress state, we first investigated nitric oxide (NO) production and cytotoxicity using Griess reagent assays, LDH assay, and neurosphere counting. Also, we investigated the effect of melatonin on NSCs by measuring the mRNA levels of SOX2, TLX, and FGFR-2. In addition, western blot analyses were performed to examine the activation of PI3K/Akt/Nrf2 signaling in LPS-treated NSCs. In the present study, we suggested that melatonin inhibits NO production and protects NSCs against LPS-induced inflammatory stress. In addition, melatonin promoted the expression of SOX2 and activated the PI3K/Akt/Nrf2 signaling under LPS-induced inflammation condition. Based on our results, we conclude that melatonin may be an important factor for the survival and proliferation of NSCs in neuroinflammatory diseases. PMID:25705693

Kang, So Mang; Lee, Kyoung Min

2015-01-01

15

The Protective Effect of Melatonin on Neural Stem Cell against LPS-Induced Inflammation.  

PubMed

Stem cell therapy for tissue regeneration has several limitations in the fact that transplanted cells could not survive for a long time. For solving these limitations, many studies have focused on the antioxidants to increase survival rate of neural stem cells (NSCs). Melatonin, an antioxidant synthesized in the pineal gland, plays multiple roles in various physiological mechanisms. Melatonin exerts neuroprotective effects in the central nervous system. To determine the effect of melatonin on NSCs which is in LPS-induced inflammatory stress state, we first investigated nitric oxide (NO) production and cytotoxicity using Griess reagent assays, LDH assay, and neurosphere counting. Also, we investigated the effect of melatonin on NSCs by measuring the mRNA levels of SOX2, TLX, and FGFR-2. In addition, western blot analyses were performed to examine the activation of PI3K/Akt/Nrf2 signaling in LPS-treated NSCs. In the present study, we suggested that melatonin inhibits NO production and protects NSCs against LPS-induced inflammatory stress. In addition, melatonin promoted the expression of SOX2 and activated the PI3K/Akt/Nrf2 signaling under LPS-induced inflammation condition. Based on our results, we conclude that melatonin may be an important factor for the survival and proliferation of NSCs in neuroinflammatory diseases. PMID:25705693

Song, Juhyun; Kang, So Mang; Lee, Kyoung Min; Lee, Jong Eun

2015-01-01

16

LPS-induced inflammatory response after therapy of aggressive periodontitis.  

PubMed

We have reported a lipopolysaccharide (LPS)-induced hyper-inflammatory response in localized aggressive periodontitis (LAP). It is unknown whether treatment is able to modulate this LPS responsiveness. Fifty-nine individuals with LAP were treated by mechanical debridement and systemic antibiotics. Clinical parameters and cyto/chemokine responsiveness of whole blood stimulated with Porphyromonas gingivalis or Escherichia coli LPS were monitored at baseline and 3, 6, and 12 months post-treatment. Overall, clinical parameters were improved following treatment. Additionally, P. gingivalis LPS induction of eotaxin, IFN?, IL10, IL12p40, IL1?, IL6, IP10, MCP1, MIP1?, GM-CSF, and TNF? was significantly decreased (p < .05). Similarly, induction of eotaxin, INF?, IL10, IL12p40, GM-CSF, and TNF? by E. coli LPS was also reduced post-treatment. These reductions correlated with decreases in clinical parameters. Importantly, these reductions in LPS responsiveness were most robust at 3 months, and some lost significance at 6 to 12 months post-treatment. In conclusion, LPS-induced hyper-inflammatory response in LAP can be partially modulated by periodontal therapy. Conversely, rebound in the hyper-responsiveness of some mediators, in the presence of improved clinical parameters, suggests that this phenotype could be partially influenced by a genetic trait and play a role in future disease recurrence (ClinicalTrials.gov, NCT01330719). PMID:23788609

Shaddox, L M; Gonçalves, P F; Vovk, A; Allin, N; Huang, H; Hou, W; Aukhil, I; Wallet, S M

2013-08-01

17

LPS-induced Inflammatory Response after Therapy of Aggressive Periodontitis  

PubMed Central

We have reported a lipopolysaccharide (LPS)-induced hyper-inflammatory response in localized aggressive periodontitis (LAP). It is unknown whether treatment is able to modulate this LPS responsiveness. Fifty-nine individuals with LAP were treated by mechanical debridement and systemic antibiotics. Clinical parameters and cyto/chemokine responsiveness of whole blood stimulated with Porphyromonas gingivalis or Escherichia coli LPS were monitored at baseline and 3, 6, and 12 months post-treatment. Overall, clinical parameters were improved following treatment. Additionally, P. gingivalis LPS induction of eotaxin, IFN?, IL10, IL12p40, IL1?, IL6, IP10, MCP1, MIP1?, GM-CSF, and TNF? was significantly decreased (p < .05). Similarly, induction of eotaxin, INF?, IL10, IL12p40, GM-CSF, and TNF? by E. coli LPS was also reduced post-treatment. These reductions correlated with decreases in clinical parameters. Importantly, these reductions in LPS responsiveness were most robust at 3 months, and some lost significance at 6 to 12 months post-treatment. In conclusion, LPS-induced hyper-inflammatory response in LAP can be partially modulated by periodontal therapy. Conversely, rebound in the hyper-responsiveness of some mediators, in the presence of improved clinical parameters, suggests that this phenotype could be partially influenced by a genetic trait and play a role in future disease recurrence (ClinicalTrials.gov, NCT01330719). PMID:23788609

Shaddox, L.M.; Gonçalves, P.F.; Vovk, A.; Allin, N.; Huang, H.; Hou, W.; Aukhil, I.; Wallet, S.M.

2013-01-01

18

Antioxidant, inhibition of ?-glucosidase and suppression of nitric oxide production in LPS-induced murine macrophages by different fractions of Actinidia arguta stem  

PubMed Central

In traditional systems of medicine, fruits, leaves, and stems of Actinidia arguta (Sieb. et Zucc.) Planch. ex Miq. have been used to treat various inflammatory diseases. The present study determined the proximate composition, antioxidant, anti-inflammatory, and hypoglycemic potential of A. arguta stem. Phenolic composition of hot water extract and its sub-fractions was determined by Folin–Ciocalteu’s reagent method. In vitro antioxidant activities of the samples were evaluated using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2?-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging assays. Anti-inflammatory activity of different fractions was investigated through the inhibition of nitric oxide (NO) production in lipopolysaccharide (1 ?g/ml) stimulated RAW 264.7 cells. In addition, inhibition of ?-glucosidase activity of hot water extract was determined using p-nitrophenyl-?-d-glucopyranoside (pNPG) as a substrate. Ethyl acetate (557.23 mg GAE/g) fraction contains higher level of total phenolic content. The antioxidant activity evaluated by DPPH radical scavenging assay showed a strong activity for ethyl acetate (IC50 of 14.28 ?g/ml) and n-butanol fractions (IC50 of 48.27 ?g/ml). Further, ethyl acetate fraction effectively inhibited NO production in RAW 264.7 cells induced by lipopolysaccharide (LPS) than other fractions (nitrite level to 32.14 ?M at 200 ?g/ml). In addition, hot water extract of A. arguta stem exhibited appreciable inhibitory activity against ?-glucosidase enzyme with IC50 of 1.71 mg/ml. The obtained results have important consequence of using A. arguta stem toward the development of effective anti-inflammatory drugs. PMID:25473361

Lee, Jaehak; Sowndhararajan, Kandhasamy; Kim, Mihae; Kim, Jaehun; Kim, Daeho; Kim, Sunpyo; Kim, Gur-Yoo; Kim, Songmun; Jhoo, Jin-Woo

2014-01-01

19

Carabrol suppresses LPS-induced nitric oxide synthase expression by inactivation of p38 and JNK via inhibition of I-{kappa}B{alpha} degradation in RAW 264.7 cells  

SciTech Connect

Carabrol, isolated from Carpesium macrocephalum, showed anti-inflammatory potential in LPS-induced RAW 264.7 murine macrophages. In present study, carabrol demonstrated the inhibitory activity on pro-inflammatory cytokines such as IL-1{beta}, IL-6 and TNF-{alpha}. In addition, mRNA and protein levels of iNOS and COX-2 were reduced by carabrol. Molecular analysis revealed that these suppressive effects were correlated with the inactivation of p38 and JNK via inhibition of NF-{kappa}B activation. Immunoblotting showed that carabrol suppressed LPS-induced degradation of I-{kappa}B{alpha} and decreased nuclear translocation of p65. Taken together, these results suggest that carabrol can be a modulator of pro-inflammatory signal transduction pathway in RAW 264.7 cells.

Lee, Hwa Jin [Department of Life Science, Research Center for Women's Diseases, Sookmyung Women's University, Seoul 140-742 (Korea, Republic of)] [Department of Life Science, Research Center for Women's Diseases, Sookmyung Women's University, Seoul 140-742 (Korea, Republic of); Lim, Hyo Jin; Lee, Da Yeon [College of Pharmacy, Sookmyung Women's University, Seoul 140-742 (Korea, Republic of)] [College of Pharmacy, Sookmyung Women's University, Seoul 140-742 (Korea, Republic of); Jung, Hyeyoun [Department of Life Science, Research Center for Women's Diseases, Sookmyung Women's University, Seoul 140-742 (Korea, Republic of)] [Department of Life Science, Research Center for Women's Diseases, Sookmyung Women's University, Seoul 140-742 (Korea, Republic of); Kim, Mi-Ran; Moon, Dong-Cheul [College of Pharmacy, Chungbuk National University, Cheungju 361-763 (Korea, Republic of)] [College of Pharmacy, Chungbuk National University, Cheungju 361-763 (Korea, Republic of); Kim, Keun Il; Lee, Myeong-Sok [Department of Life Science, Research Center for Women's Diseases, Sookmyung Women's University, Seoul 140-742 (Korea, Republic of)] [Department of Life Science, Research Center for Women's Diseases, Sookmyung Women's University, Seoul 140-742 (Korea, Republic of); Ryu, Jae-Ha, E-mail: ryuha@sookmyung.ac.kr [College of Pharmacy, Sookmyung Women's University, Seoul 140-742 (Korea, Republic of)] [College of Pharmacy, Sookmyung Women's University, Seoul 140-742 (Korea, Republic of)

2010-01-15

20

Amelioration of LPS-Induced Inflammation Response in Microglia by AMPK Activation  

PubMed Central

Adenosine 5?-monophosphate-activated protein kinase (AMPK) is a key regulator of cellular energy homeostasis via modulating metabolism of glucose, lipid, and protein. In addition to energy modulation, AMPK has been demonstrated to associate with several important cellular events including inflammation. The results showed that ENERGI-F704 identified from bamboo shoot extract was nontoxic in concentrations up to 80??M and dose-dependently induced phosphorylation of AMPK (Thr-172) in microglia BV2 cells. Our findings also showed that the treatment of BV2 with ENERGI-F704 ameliorated the LPS-induced elevation of IL-6 and TNF-? production. In addition, ENERGI-F704 reduced increased production of nitric oxide (NO) and prostaglandin E2 (PGE2) via downregulating the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2), respectively. Moreover, ENERGI-F704 decreased activated nuclear translocation and protein level of NF-?B. Inhibition of AMPK with compound C restored decreased NF-?B translocation by ENERGI-F704. In conclusion, ENERGI-F704 exerts inhibitory activity on LPS-induced inflammation through manipulating AMPK signaling and exhibits a potential therapeutic agent for neuroinflammatory disease. PMID:25025067

Chen, Chin-Chen; Lin, Jiun-Tsai; Cheng, Yi-Fang; Kuo, Cheng-Yi; Huang, Chun-Fang; Kao, Shao-Hsuan; Liang, Yao-Jen; Cheng, Ching-Yi; Chen, Han-Min

2014-01-01

21

LPS induces pulp progenitor cell recruitment via complement activation.  

PubMed

Complement system, a major component of the natural immunity, has been recently identified as an important mediator of the dentin-pulp regeneration process through STRO-1 pulp cell recruitment by the C5a active fragment. Moreover, it has been shown recently that under stimulation with lipoteichoic acid, a complex component of the Gram-positive bacteria cell wall, human pulp fibroblasts are able to synthesize all proteins required for complement activation. However, Gram-negative bacteria, which are also involved in tooth decay, are known as powerful activators of complement system and inflammation. Here, we investigated the role of Gram-negative bacteria-induced complement activation on the pulp progenitor cell recruitment using lipopolysaccharide (LPS), a major component of all Gram-negative bacteria. Our results show that incubating pulp fibroblasts with LPS induced membrane attack complex formation and C5a release in serum-free fibroblast cultures. The produced C5a binds to the pulp progenitor cells' membrane and induces their migration toward the LPS stimulation chamber, as revealed by the dynamic transwell migration assays. The inhibition of this migration by the C5aR-specific antagonist W54011 indicates that the pulp progenitor migration is mediated by the interaction between C5a and C5aR. Our findings demonstrate, for the first time, a direct interaction between the recruitment of progenitor pulp cells and the activation of complement system generated by pulp fibroblast stimulation with LPS. PMID:25359783

Chmilewsky, F; Jeanneau, C; Laurent, P; About, I

2015-01-01

22

Acanthoic Acid inhibits LPS-induced inflammatory response in human gingival fibroblasts.  

PubMed

Periodontitis is a chronic disease that affects the gums and destroys connective tissue. Acanthoic acid (AA), a diterpene in Acanthopanax koreanum, has been reported to have anti-inflammatory activities. The aim of this study was to investigate the anti-inflammatory effects of AA on lipopolysaccharide (LPS)-induced inflammatory response in human gingival fibroblasts (HGFs). HGFs were treated with Porphyromonas gingivalis LPS in the presence or absence of AA. The production of inflammatory cytokines IL-8 and IL-6 were measured by ELISA. The expression of NF-?B and TLR4 were detected by Western blotting. The results showed that AA inhibited LPS-induced IL-8 and IL-6 production in a dose-dependent manner. In addition, AA inhibited LPS-induced TLR4 expression and NF-?B activation. In conclusion, AA inhibits LPS-induced inflammatory response in HGFs through inhibition TLR4-mediated NF-?B signaling pathway. PMID:25373915

Wei, Cai; Tan, Chee Keong; Xiaoping, He; Junqiang, Jiang

2015-04-01

23

MEMANTINE PROTECTS AGAINST LPS-INDUCED NEUROINFLAMMATION, RESTORES BEHAVIORALLY-INDUCED GENE  

E-print Network

MEMANTINE PROTECTS AGAINST LPS-INDUCED NEUROINFLAMMATION, RESTORES BEHAVIORALLY-INDUCED GENE of Arc, and c) spatial memory deficits. Memantine, a low to moderate affinity open channel uncompetitive and memantine (10 mg/kg/day memantine s.c.). The re- sults reported here demonstrate that memantine reduces OX6

Wenk, Gary

24

Paroxetine differentially modulates LPS-induced TNF? and IL-6 production in mouse macrophages.  

PubMed

Paroxetine is a selective serotonin reuptake inhibitor (SSRI) that is clinically used for the treatment of depression in human patients. Because of recent reports on the role of serotonin in modulating inflammation and the link between inflammation and depression, we sought to test the effect of paroxetine directly on macrophage response to an inflammatory stimulus. Lipopolysaccharide (LPS) treatment of mouse macrophages significantly enhanced TNF? and IL-6 production. Paroxetine treatment of macrophages, however, significantly inhibited LPS-induced IL-6 production. In contrast, paroxetine enhanced LPS-induced TNF? production in macrophages. These effects of paroxetine were mimicked by fluoxetine, another SSRI. To determine if the effects of paroxetine are mediated via modulation of the 5-HT system, we treated macrophages with 5-HT or 5-HT receptor antagonist (LY215840) in the presence of LPS and/or paroxetine. 5-HT treatment by itself did not affect LPS-induced cytokine production. LY215840, however, reversed paroxetine's effect on LPS-induced TNF? production but not IL-6. To understand the signaling mechanisms, we examined paroxetine's effect on MAPK and NF?B pathways. While paroxetine inhibited LPS-induced I?B? phosphorylation, MAPK pathways were mostly unaffected. Together these data demonstrate that paroxetine has critical but differential effects on IL-6 and TNF? production in macrophages and that it likely regulates these cytokines via distinct mechanisms. PMID:25744603

Durairaj, Haritha; Steury, Michael D; Parameswaran, Narayanan

2015-04-01

25

Attenuation of lipopolysaccharide (LPS)-induced cytotoxicity by tocopherols and tocotrienols?  

PubMed Central

Lipopolysaccharide (LPS) induces host inflammatory responses and tissue injury and has been implicated in the pathogenesis of various age-related diseases such as acute respiratory distress syndrome, vascular diseases, and periodontal disease. Antioxidants, particularly vitamin E, have been shown to suppress oxidative stress induced by LPS, but the previous studies with different vitamin E isoforms gave inconsistent results. In the present study, the protective effects of ?- and ?-tocopherols and ?- and ?-tocotrienols on the oxidative stress induced by LPS against human lung carcinoma A549 cells were studied. They suppressed intracellular reactive oxygen formation, lipid peroxidation, induction of inflammatory mediator cytokines, and cell death. Tocopherols were incorporated into cultured cells much slower than tocotrienols but could suppress LPS-induced oxidative stress at much lower intracellular concentration than tocotrienols. Considering the bioavailability, it was concluded that ?-tocopherol may exhibit the highest protective capacity among the vitamin E isoforms against LPS-induced oxidative stress. PMID:24024142

Nishio, Keiko; Horie, Masanori; Akazawa, Yoko; Shichiri, Mototada; Iwahashi, Hitoshi; Hagihara, Yoshihisa; Yoshida, Yasukazu; Niki, Etsuo

2013-01-01

26

A central role for the mammalian target of rapamycin in LPS-induced anorexia in mice.  

PubMed

Bacterial lipopolysaccharide (LPS), also known as endotoxin, induces profound anorexia. However, the LPS-provoked pro-inflammatory signaling cascades and the neural mechanisms underlying the development of anorexia are not clear. Mammalian target of rapamycin (mTOR) is a key regulator of metabolism, cell growth, and protein synthesis. This study aimed to determine whether the mTOR pathway is involved in LPS-induced anorexia. Effects of LPS on hypothalamic gene/protein expression in mice were measured by RT-PCR or western blotting analysis. To determine whether inhibition of mTOR signaling could attenuate LPS-induced anorexia, we administered an i.c.v. injection of rapamycin, an mTOR inhibitor, on LPS-treated male mice. In this study, we showed that LPS stimulates the mTOR signaling pathway through the enhanced phosphorylation of mTOR(Ser2448) and p70S6K(Thr389). We also showed that LPS administration increased the phosphorylation of FOXO1(Ser256), the p65 subunit of nuclear factor kappa B (P<0.05), and FOXO1/3a(Thr) (24) (/) (32) (P<0.01). Blocking the mTOR pathway significantly attenuated the LPS-induced anorexia by decreasing the phosphorylation of p70S6K(Thr389), FOXO1(Ser256), and FOXO1/3a(Thr) (24) (/) (32). These results suggest promising approaches for the prevention and treatment of LPS-induced anorexia. PMID:25349249

Yue, Yunshuang; Wang, Yi; Li, Dan; Song, Zhigang; Jiao, Hongchao; Lin, Hai

2015-01-01

27

Low-Level Laser Therapy Attenuates LPS-Induced Rats Mastitis by Inhibiting Polymorphonuclear Neutrophil Adhesion  

PubMed Central

ABSTRACT The aim of this study was to investigate the effects of low-level laser therapy (LLLT) on a rat model of lipopolysaccharide (LPS)-induced mastitis and its underlying molecular mechanisms. The rat model of mastitis was induced by inoculation of LPS through the canals of the mammary gland. The results showed that LPS-induced secretion of IL-1? and IL-8 significantly decreased after LLLT (650 nm, 2.5 mW, 30 mW/cm2). LLLT also inhibited intercellular adhesion molecule-1 (ICAM-1) expression and attenuated the LPS-induced decrease of the expression of CD62L and increase of the expression of CD11b. Moreover, LLLT also suppressed LPS-induced polymorphonuclear neutrophils (PMNs) entering the alveoli of the mammary gland. The number of PMNs in the mammary alveolus and the myeloperoxidase (MPO) activity were decreased after LLLT. These results suggested that LLLT therapy is beneficial in decreasing the somatic cell count and improving milk nutritional quality in cows with an intramammary infection. PMID:25452258

WANG, Yueqiang; HE, Xianjing; HAO, Dandan; YU, Debin; LIANG, Jianbin; QU, Yanpeng; SUN, Dongbo; YANG, Bin; YANG, Keli; WU, Rui; WANG, Jianfa

2014-01-01

28

Protective effect of taraxasterol against LPS-induced endotoxic shock by modulating inflammatory responses in mice.  

PubMed

Taraxasterol, a pentacyclic-triterpene, was isolated from the Chinese medicinal herb Taraxacum officinale. In the present study, we investigated the protective effect of taraxasterol on murine model of endotoxic shock and the mechanism of its action. Mice were treated with 2.5, 5 and 10 mg/kg of taraxasterol prior to a lethal dose of lipopolysaccharide (LPS) challenge. Survival of mice was monitored twice a day for 7 days. To further understand the mechanism, the serum levels of inflammatory cytokine tumor necrosis factor-? (TNF-?), interferon-? (IFN-?), interleukin-1? (IL-1?), interleukin-6 (IL-6) and mediator nitric oxide (NO), prostaglandin E? (PGE?) as well as histology of lungs were examined. The results showed that taraxasterol significantly improved mouse survival and attenuated tissue injury of the lungs in LPS-induced endotoxemic mice. Further studies revealed that taraxasterol significantly reduced TNF-?, IFN-?, IL-1?, IL-6, NO and PGE? levels in sera from mice with endotoxic shock. These results indicate that taraxasterol has a protective effect on murine endotoxic shock induced by LPS through modulating inflammatory cytokine and mediator secretion. This finding might provide a new strategy for the treatment of endotoxic shock and associated inflammation. PMID:24286370

Zhang, Xuemei; Xiong, Huanzhang; Li, Hongyu; Cheng, Yao

2014-02-01

29

EFFECTS OF SYSTEMIC NEUTROPHIL DEPLETION ON LPS-INDUCED AIRWAY DISEASE  

EPA Science Inventory

Effects of Systemic Neutrophil Depletion on LPS-induced Airway Disease Jordan D. Savov, Stephen H. Gavett*, David M. Brass, Daniel L. Costa*, David A. Schwartz Pulmonary and Critical Care Division, Dept of Medicine ? Duke University Medical Center * National Health and E...

30

NEUTROPHILS PLAY A CRITICAL ROLE IN THE DEVELOPMENT OF LPS-INDUCED AIRWAY DISEASE  

EPA Science Inventory

ETD-02-045 (GAVETT) GPRA # 10108 Neutrophils Play a Critical Role in the Development of LPS-Induced Airway Disease. Jordan D. Savov, Stephen H. Gavett*, David M. Brass, Daniel L. Costa*, and David A. Schwartz ABSTRACT We investigated the role of neutrophils...

31

LPS INDUCES IL6 IN THE BRAIN AND IN SERUM LARGELY THROUGH TNF PRODUCTION  

Microsoft Academic Search

We investigated the relative contribution of IL-6 and PGE2 directly induced by LPS and indirectly induced via TNF, using in vivo and in vitro models in the mouse. In these models we have used as tools an anti-TNF antibody and a cyclooxygenase inhibitor, the S enantiomer of ketoprofen (S-KPF). Anti-TNF antibodies inhibited LPS-induced IL-6 production in three different models: IL-6

Pietro Ghezzi; Silvano Sacco; Davide Agnello; Antonello Marullo; Gianfranco Caselli; Riccardo Bertini

2000-01-01

32

Angiopoietin-1 variant reduces LPS-induced microvascular dysfunction in a murine model of sepsis  

PubMed Central

Introduction Severe sepsis is characterised by intravascular or extravascular infection with microbial agents, systemic inflammation and microcirculatory dysfunction, leading to tissue damage, organ failure and death. The growth factor angiopoietin (Ang-1) has therapeutic potential but recombinant Ang-1 tends to aggregate and has a short half-life in vivo. This study aimed to investigate the acute effects of the more stable Ang-1 variant matrilin-1-angiopoietin-1 (MAT.Ang-1) on the function of the microcirculation in an experimental model of sepsis, and whether any protection by MAT-Ang-1 was associated with modulation of inflammatory cytokines, angiogenic factors or the endothelial nitric oxide synthase (eNOS)-Akt and vascular endothelial (VE)-cadherin pathways. Methods Aluminium window chambers were implanted into the dorsal skinfold of male C3H/HeN mice (7 to 10 weeks old) to expose the striated muscle microcirculation. Endotoxemia was induced by intraperitoneal injection of lipopolysaccharide (LPS, 1 mg/kg at 0 and 19 hours). MAT.Ang-1 was administered intravenously 20 hours after the onset of sepsis. Microcirculatory function was evaluated by intravital microscopy and Doppler fluximetry. Results Endotoxemia resulted in macromolecular leak, which was ameliorated by MAT.Ang-1 post-treatment. LPS induced a dramatic reduction in tissue perfusion, which was improved by MAT.Ang-1. Proteome profiler array analysis of skeletal muscle also demonstrated increased inflammatory and reduced angiogenic factors during endotoxemia. MAT.Ang-1 post-treatment reduced the level of IL-1? but did not significantly induce the expression of angiogenic factors. MAT.Ang-1 alone did not induce leak or increase angiogenic factors but did reduce vascular endothelial growth factor expression in controls. Conclusion Administration of MAT.Ang-1 after the onset of sepsis protects the microcirculation from endotoxemia-induced vascular dysfunction through reducing inflammation but without pro-angiogenic actions, thus representing a novel, potential pharmacotherapeutic agent for the treatment of sepsis. PMID:23036162

2012-01-01

33

Effects of cannabinoid receptor ligands on LPS-induced pulmonary inflammation in mice  

Microsoft Academic Search

The effects of cannabinoid receptor agonists WIN 55,212-2, ?9-tetrahydrocannabinol (?9-THC), arachidonoylethanolamide (anandamide) and palmitoylethanolamide on lipopolysaccharide (LPS) -induced bronchopulmonary inflammation in mice were investigated. WIN 55,212-2 and ?9-THC induced a concentration-dependent decrease in TNF-? level in the bronchoalveolar lavage fluid (BALF) (maximum inhibition 52.7% and 36.9% for intranasal doses of 750 nmol.kg?1 and 2.65 mmol.kg?1, respectively). This effect was accompanied

E. Berdyshev; E. Boichot; M. Corbel; N. Germain; V Lagente

1998-01-01

34

Protection against LPS-induced cartilage inflammation and degradation provided by a biological extract of Mentha spicata  

PubMed Central

Background A variety of mint [Mentha spicata] has been bred which over-expresses Rosmarinic acid (RA) by approximately 20-fold. RA has demonstrated significant anti-inflammatory activity in vitro and in small rodents; thus it was hypothesized that this plant would demonstrate significant anti-inflammatory activity in vitro. The objectives of this study were: a) to develop an in vitro extraction procedure which mimics digestion and hepatic metabolism, b) to compare anti-inflammatory properties of High-Rosmarinic-Acid Mentha spicata (HRAM) with wild-type control M. spicata (CM), and c) to quantify the relative contributions of RA and three of its hepatic metabolites [ferulic acid (FA), caffeic acid (CA), coumaric acid (CO)] to anti-inflammatory activity of HRAM. Methods HRAM and CM were incubated in simulated gastric and intestinal fluid, liver microsomes (from male rat) and NADPH. Concentrations of RA, CA, CO, and FA in simulated digest of HRAM (HRAMsim) and CM (CMsim) were determined (HPLC) and compared with concentrations in aqueous extracts of HRAM and CM. Cartilage explants (porcine) were cultured with LPS (0 or 3 ?g/mL) and test article [HRAMsim (0, 8, 40, 80, 240, or 400 ?g/mL), or CMsim (0, 1, 5 or 10 mg/mL), or RA (0.640 ?g/mL), or CA (0.384 ?g/mL), or CO (0.057 ?g/mL) or FA (0.038 ?g/mL)] for 96 h. Media samples were analyzed for prostaglandin E2 (PGE2), interleukin 1? (IL-1), glycosaminoglycan (GAG), nitric oxide (NO) and cell viability (differential live-dead cell staining). Results RA concentration of HRAMsim and CMsim was 49.3 and 0.4 ?g/mL, respectively. CA, FA and CO were identified in HRAMsim but not in aqueous extract of HRAM. HRAMsim (? 8 ?g/mL) inhibited LPS-induced PGE2 and NO; HRAMsim (? 80 ?g/mL) inhibited LPS-induced GAG release. RA inhibited LPS-induced GAG release. No anti-inflammatory or chondroprotective effects of RA metabolites on cartilage explants were identified. Conclusions Our biological extraction procedure produces a substance which is similar in composition to post-hepatic products. HRAMsim is an effective inhibitor of LPS-induced inflammation in cartilage explants, and effects are primarily independent of RA. Further research is needed to identify bioactive phytochemical(s) in HRAMsim. PMID:20459798

2010-01-01

35

Nucleotide receptor P2RX7 stimulation enhances LPS-induced interferon-? production in murine macrophages.  

PubMed

Stimulation of P2RX(7) with extracellular ATP potentiates numerous LPS-induced proinflammatory events, including cytokine induction in macrophages, but the molecular mechanisms underlying this process are not well defined. Although P2RX(7) ligation has been proposed to activate several transcription factors, many of the LPS-induced mediators affected by P2RX(7) activation are not induced by P2RX(7) agonists alone, suggesting a complementary role for P2RX(7) in transcriptional regulation. Type I IFN production, whose expression is tightly controlled by multiple transcription factors that form an enhanceosome, is critical for resistance against LPS-containing bacteria. The effect of purinergic receptor signaling on LPS-dependent type I IFN is unknown and would be of great relevance to a diverse array of inflammatory conditions. The present study demonstrates that stimulation of macrophages with P2RX(7) agonists substantially enhances LPS-induced IFN-? expression, and this enhancement is ablated in macrophages that do not express functional P2RX(7) or when the MAPK MEK1/2 pathways are inhibited. Potentiation of LPS-induced IFN-? expression following P2RX(7) stimulation is likely transcriptionally regulated, as this enhancement is observed at the IFN-? promoter level. Furthermore, P2RX(7) stimulation is able to increase the phosphorylation and subsequent IFN-? promoter occupancy of IRF-3, a transcription factor that is critical for IFN-? transcription by TLR agonists. This newly discovered role for P2RX(7) in IFN regulation may have implications in antimicrobial defense, which has been linked to P2RX(7) activation in other studies. PMID:23911869

Gavala, M L; Liu, Y-P; Lenertz, L Y; Zeng, L; Blanchette, J B; Guadarrama, A G; Denlinger, L C; Bertics, P J; Smith, J A

2013-10-01

36

Impact of LPS-induced cardiomyoblast cell apoptosis inhibited by earthworm extracts.  

PubMed

Dilong is an earthworm extract with a dense nutritional content, widely used in Chinese herbal medicine to remove stasis and stimulate wound healing. Earthworm extracts are traditionally used by indigenous people throughout the world. How this Dilong inhibits Lipopolysaccharide (LPS)-induced cardiomyoblast cell apoptosis is still unclear. This study investigates the Dilong extract effect on LPS-induced apoptosis in H9c2 cardiomyoblast cells. LPS (1 ?g/ml) administration for 24 h induced apoptosis in H9c2 cells. Cell apoptosis was detected using MTT, LDH, TUNEL assay and JC-1 staining. Western blot analysis was used to detect pro-apoptotic and anti-apoptotic proteins. Dilong extract totally blocked the LPS impact, leading to the activation of anti-apoptotic proteins, Bcl-2 and Bcl-xL, stabilized the mitochondria membrane and down-regulated the extrinsic and intrinsic pro-apoptotic proteins, TNF-?, active caspase-8, t-Bid, Bax, active caspase-9 and active caspase-3. Dilong could potentially serve as a cardio protective agent against LPS-induced H9c2 cardiomyoblast cell apoptosis. PMID:25249212

Li, Ping-Chun; Tien, Yun-Chen; Day, Cecilia Hsuan; Pai, Peiying; Kuo, Wei-Wen; Chen, Tung-Sheng; Kuo, Chia-Hua; Tsai, Chang-Hai; Ju, Da-Tong; Huang, Chih-Yang

2015-04-01

37

Neuraminidase reprograms lung tissue and potentiates LPS-induced acute lung injury in mice  

PubMed Central

We previously reported that removal of sialyl residues primed PBMCs to respond to bacterial LPS stimulation in vitro. Therefore, we speculated that prior desialylation can sensitize the host to generate an enhanced inflammatory response upon exposure to a TLR ligand, such as LPS, in a murine model of acute lung injury. Intratracheal instillation of neuraminidase (NA) 30 min prior to intratracheal administration of LPS increased PMNs in the bronchoalveolar lavage fluid (BALF) and the wet-to-dry lung weight ratio, a measure of pulmonary edema, compared to mice that received LPS alone. Administration of NA alone resulted in desialylation of bronchiolar and alveolar surfaces and induction of TNF-?, IL-1?, and chemokines in lung homogenates and BALF; however, PMN recruitment in mice treated with NA alone did not differ from those of PBS-administered controls. NA pretreatment alone induced apoptosis and markedly enhanced LPS-induced endothelial apoptosis. Administration of recombinant Bcl-2, an anti-apoptotic molecule, abolished the effect of NA treatment on LPS-induced PMN recruitment and pulmonary edema formation. We conclude that NA pretreatment potentiates LPS-induced lung injury through enhanced PMN recruitment, pulmonary edema formation, and endothelial and myeloid cell apoptosis. A similar “reprogramming” of immune responses with desialylation may occur during respiratory infection with NA-expressing microbes and contribute to severe lung injury. PMID:24068662

Feng, Chiguang; Zhang, Lei; Nguyen, Chinh; Vogel, Stefanie N.; Goldblum, Simeon E.; Blackwelder, William C.; Cross, Alan S.

2013-01-01

38

LPS-induced biomarkers in mice: A potential model for identifying insulin sensitizers  

Microsoft Academic Search

The contribution of nutrient overload and associated inflammation to insulin resistance has highlighted several therapeutic targets including c-Jun N-terminal kinase (JNK) and S6 kinase (S6K). To investigate how a lipopolysaccharide (LPS)-mediated inflammatory response may modulate pathways implicated in insulin resistance, we characterized the LPS-induced changes in key biomarkers. Administration of 0.06–4mg\\/kg LPS to C57BL\\/6 mice stimulated increases in plasma levels

Celia P. Briscoe; David Looper; Phong Tran; Jocelyn Herrera; Scott R. McDonnell; B. Ganesh Bhat

2007-01-01

39

Tim-3 Negatively Mediates Natural Killer Cell Function in LPS-Induced Endotoxic Shock  

PubMed Central

Sepsis is an exaggerated inflammatory condition response to different microorganisms with high mortality rates and extremely poor prognosis. Natural killer (NK) cells have been reported to be the major producers of IFN-? and key players in promoting systematic inflammation in lipopolysaccharide (LPS)-induced endotoxic shock. T-cell immunoglobulin and mucin domain (Tim)-3 pathway has been demonstrated to play an important role in the process of sepsis, however, the effect of Tim-3 on NK cell function remains largely unknown. In this study, we observed a dynamic inverse correlation between Tim-3 expression and IFN-? production in NK cells from LPS-induced septic mice. Blockade of the Tim-3 pathway could increase IFN-? production and decrease apoptosis of NK cells in vitro, but had no effect on the expression of CD107a. Furthermore, NK cell cytotoxicity against K562 target cells was enhanced after blocking Tim-3 pathway. In conclusion, our results suggest that Tim-3 pathway plays an inhibitory role in NK cell function, which might be a potential target in modulating the excessive inflammatory response of LPS-induced endotoxic shock. PMID:25337993

Hou, Hongyan; Liu, Weiyong; Wu, Shiji; Lu, Yanjun; Peng, Jing; Zhu, Yaowu; Lu, Yanfang; Wang, Feng; Sun, Ziyong

2014-01-01

40

TLR4 Mediates LPS-Induced VEGF Expression in Odontoblasts  

Microsoft Academic Search

Lipopolysaccharide (LPS) from gram-negative bacteria cell walls such as Prevotella intermedia and Escherichia coli induce vascular endothelial growth factor (VEGF) expression in odontoblasts, but not in undifferentiated dental pulp cells. CD14 and TLR4 are responsible for LPS signaling in macrophages, but their expression levels and function in dental pulp cells are unknown. We showed here that murine odontoblast-like cells (MDPC-23)

Tatiana M. Botero; Charles E. Shelburne; G. Rex Holland; Carl T. Hanks; Jacques E. Nör

2006-01-01

41

Wedelolactone inhibits LPS-induced pro-inflammation via NF-kappaB Pathway in RAW 264.7 cells  

PubMed Central

Background Wedelolactone (WEL), a major coumestan ingredient in Wedelia chinensis, has been used to treat septic shock, hepatitis and venom poisoning in traditional Chinese medicines. The objective of the study was to elucidate the anti-inflammatory effects and mechanism of WEL with a cellular model of lipopolysaccharide (LPS)-induced RAW 264.7 cells. Results To study the role of WEL in pro-inflammation, we measured key inflammation mediators and end products including nitric oxide (NO), prostaglandin E2 (PGE2), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and tumor necrosis factor-? (TNF-?) by using the Griess method, enzyme linked immunosorbent assay (ELISA) and Western blotting. Nuclear factor-kappaB (NF-?B) transcription activity was detected by luciferase reporter assay. The important pro-inflammatory transcription factors, NF-?B p65 and inhibitory kappaB alpha (I?B-?); and mitogen-activated protein kinases (MAPKs), including extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 MAPK (p38) were analyzed by Western blotting. Our study showed that WEL (0.1, 1, 10 ?M) significantly inhibited the protein expression levels of iNOS and COX-2 in LPS-stimulated cells, as well as the downstream products, including NO, PGE2 and TNF-?. Moreover, WEL also inhibited LPS-induced NF-?B p65 activation via the degradation and phosphorylation of I?B-? and subsequent translocation of the NF-?B p65 subunit to the nucleus. Conclusions Our results revealed that WEL has a potential to be a novel anti-inflammatory agent targeting on the NF-?B signaling pathway. PMID:24176090

2013-01-01

42

Manganese Potentiates LPS-Induced Heme-Oxygenase 1 in Microglia but not Dopaminergic Cells: Role in Controlling Microglial Hydrogen Peroxide and Inflammatory Cytokine Output  

PubMed Central

Excessive manganese (Mn) exposure increases output of glial-derived inflammatory products, which may indirectly contribute to the neurotoxic effects of this essential metal. In microglia, Mn increases hydrogen peroxide (H2O2) release and potentiates lipopolysaccharide (LPS)-induced cytokines (TNF-?, IL-6) and nitric oxide (NO). Inducible heme-oxygenase (HO-1) plays a role in the regulation of inflammation and its expression is upregulated in response to oxidative stressors, including metals and LPS. Because Mn can oxidatively affect neurons both directly and indirectly, we investigated the effect of Mn exposure on the induction of HO-1 in resting and LPS-activated microglia (N9) and dopaminergic neurons (N27). In microglia, 24 h exposure to Mn (up to 250 ?M) had minimal effects on its own, but it markedly potentiated LPS (100 ng/ml)-induced HO-1protein and mRNA. Inhibition of microglial HO-1 activity with two different inhibitors indicated that HO-1 is a positive regulator of the Mn-potentiated cytokine output and a negative regulator of the Mn-induced H2O2 output. Mn enhancement of LPS-induced HO-1 does not appear to be dependent on H2O2 or NO, as Mn+LPS-induced H2O2 release was not greater than the increase induced by Mn alone and inhibition of iNOS did not change Mn potentiation of HO-1. However, because Mn exposure potentiated the LPS-induced nuclear expression of small Maf proteins, this may be one mechanism Mn uses to affect the expression of HO-1 in activated microglia. Finally, the potentiating effects of Mn on HO-1 appear to be glia-specific for Mn, LPS, or Mn+LPS did not induce HO-1 in N27 neuronal cells. PMID:21963524

Dodd, Celia A.; Filipov, Nikolay M.

2012-01-01

43

Critical role of endothelial CXCR2 in LPS-induced neutrophil migration into the lung.  

PubMed

In models of acute lung injury, CXC chemokine receptor 2 (CXCR2) mediates migration of polymorphonuclear leukocytes (PMNs) into the lung. Since CXCR2 ligands, including CXCL1 and CXCL2/3, are chemotactic for PMNs, CXCR2 is thought to recruit PMNs by inducing chemotactic migration. In a model of PMN recruitment to the lung, aerosolized bacterial LPS inhalation induced PMN recruitment to the lung in wild-type mice, but not in littermate CXCR2-/- mice. Surprisingly, lethally irradiated wild-type mice reconstituted with CXCR2-/- BM still showed about 50% PMN recruitment into bronchoalveolar lavage fluid and into lung interstitium, but CXCR2-/- mice reconstituted with CXCR2-/- BM showed no PMN recruitment. Conversely, CXCR2-/- mice reconstituted with wild-type BM showed a surprisingly large defect in PMN recruitment, inconsistent with a role of CXCR2 on PMNs alone. Cell culture, immunohistochemistry, flow cytometry, and real-time RT-PCR were used to show expression of CXCR2 on pulmonary endothelial and bronchial epithelial cells. The LPS-induced increase in lung microvascular permeability as measured by Evans blue extravasation required CXCR2 on nonhematopoietic cells. Our data revealed what we believe to be a previously unrecognized role of endothelial and epithelial CXCR2 in LPS-induced PMN recruitment and lung injury. PMID:16485040

Reutershan, Jörg; Morris, Margaret A; Burcin, Tracy L; Smith, David F; Chang, Daniel; Saprito, Mary S; Ley, Klaus

2006-03-01

44

Critical role of endothelial CXCR2 in LPS-induced neutrophil migration into the lung  

PubMed Central

In models of acute lung injury, CXC chemokine receptor 2 (CXCR2) mediates migration of polymorphonuclear leukocytes (PMNs) into the lung. Since CXCR2 ligands, including CXCL1 and CXCL2/3, are chemotactic for PMNs, CXCR2 is thought to recruit PMNs by inducing chemotactic migration. In a model of PMN recruitment to the lung, aerosolized bacterial LPS inhalation induced PMN recruitment to the lung in wild-type mice, but not in littermate CXCR2–/– mice. Surprisingly, lethally irradiated wild-type mice reconstituted with CXCR2–/– BM still showed about 50% PMN recruitment into bronchoalveolar lavage fluid and into lung interstitium, but CXCR2–/– mice reconstituted with CXCR2–/– BM showed no PMN recruitment. Conversely, CXCR2–/– mice reconstituted with wild-type BM showed a surprisingly large defect in PMN recruitment, inconsistent with a role of CXCR2 on PMNs alone. Cell culture, immunohistochemistry, flow cytometry, and real-time RT-PCR were used to show expression of CXCR2 on pulmonary endothelial and bronchial epithelial cells. The LPS-induced increase in lung microvascular permeability as measured by Evans blue extravasation required CXCR2 on nonhematopoietic cells. Our data revealed what we believe to be a previously unrecognized role of endothelial and epithelial CXCR2 in LPS-induced PMN recruitment and lung injury. PMID:16485040

Reutershan, Jörg; Morris, Margaret A.; Burcin, Tracy L.; Smith, David F.; Chang, Daniel; Saprito, Mary S.; Ley, Klaus

2006-01-01

45

The procyanidin trimer C1 inhibits LPS-induced MAPK and NF-?B signaling through TLR4 in macrophages.  

PubMed

Natural products and dietary components rich in polyphenols have been shown to reduce inflammation; however, the molecular mechanisms underlying this anti-inflammatory activity are not completely characterized, and many features remain to be elucidated. This research was carried out to clarify the potential role of procyanidin trimer C1 in the anti-inflammatory effect of polyphenols. Procyanidin C1 inhibited inducible nitric oxide synthase-mediated nitric oxide production and the release of pro-inflammatory cytokines (interleukin-6 and tumor necrosis factor-?) in lipopolysaccharide (LPS)-induced macrophages. Treatment with procyanidin C1 resulted in a significant decrease in prostaglandin E2 and cyclooxygenase-2 levels, as well as the expression of cell surface molecules (CD80, CD86, and MHC class II), which was induced by LPS. Furthermore, our data demonstrated that the anti-inflammatory effect of procyanidin C1 occurs through inhibition of mitogen-activated protein kinase (p38 and c-Jun N-terminal kinase) and nuclear factor-?B signaling pathways. These 2 factors play a major role in controlling inflammation, through toll-like receptor 4, suggesting that procyanidin C1 plays a potent role in promoting anti-inflammatory activity in macrophages. These results represent a novel and effective therapeutic intervention for the treatment of inflammatory disease. PMID:23261363

Byun, Eui-Baek; Sung, Nak-Yun; Byun, Eui-Hong; Song, Du-Sup; Kim, Jae-Kyung; Park, Jong-Heum; Song, Beom-Seok; Park, Sang-Hyun; Lee, Ju-Woon; Byun, Myung-Woo; Kim, Jae-Hun

2013-02-01

46

3,4,5-Trihydroxycinnamic Acid Inhibits LPS-Induced iNOS Expression by Suppressing NF-?B Activation in BV2 Microglial Cells  

PubMed Central

Although various derivatives of caffeic acid have been reported to possess a wide variety of biological activities such as neuronal protection against excitotoxicity and anti-inflammatory property, the biological activity of 3,4,5-trihydroxycinnamic acid (THC), a derivative of hydroxycinnamic acids, has not been clearly examined. The objective of the present study is to evaluate the anti-inflammatory effects of THC on lipopolysaccharide (LPS)-stimulated BV2 microglial cells. THC significantly suppressed LPS-induced excessive production of nitric oxide (NO) and expression of iNOS, which is responsible for the production of iNOS. THC also suppressed LPS-induced overproduction of pro-inflammatory cytokines such as IL-1? and TNF-? in BV2 microgilal cells. Furthermore, THC significantly suppressed LPS-induced degradation of I?B, which retains NF-?B in the cytoplasm. Therefore, THC attenuated nuclear translocation of NF-?B, a major pro-inflammatory transcription factor. Taken together, the present study for the first time demonstrates that THC exhibits anti-inflammatory activity through the suppression of NF-?B transcriptional activation in LPS-stimulated BV2 microglial cells. PMID:22563255

Lee, Jae-Won; Bae, Chang Jun; Choi, Yong-Jun; Kim, Song-In; Kim, Nam-Ho; Lee, Hee Jae; Kim, Sung-Soo; Kwon, Yong-Soo

2012-01-01

47

Calorie restriction attenuates lipopolysaccharide (LPS)-induced microglial activation in discrete regions of the hypothalamus and the subfornical organ.  

PubMed

Calorie restriction (CR) has been shown to increase longevity and elicit many health promoting benefits including delaying immunosenescence and attenuating neurodegeneration in animal models of Alzheimer's disease and Parkinson's disease. CR also suppresses microglial activation following cortical injury and aging. We previously demonstrated that CR attenuates lipopolysaccharide (LPS)-induced fever and shifts hypothalamic signaling pathways to an anti-inflammatory bias; however, the effects of CR on LPS-induced microglial activation remain largely unexplored. The current study investigated regional changes in LPS-induced microglial activation in mice exposed to 50% CR for 28days. Immunohistochemistry was conducted to examine changes in ionized calcium-binding adapter molecule-1 (Iba1), a protein constitutively expressed by microglia, in a total of 27 brain regions involved in immunity, stress, and/or thermoregulation. Exposure to CR attenuated LPS-induced fever, and LPS-induced microglial activation in a subset of regions: the arcuate nucleus (ARC) and ventromedial nucleus of the hypothalamus (VMH) and the subfornical organ (SFO). Microglial activation in the ARC and VMH was positively correlated with body temperature. These data suggest that CR exerts effects on regionally specific populations of microglia; particularly, in appetite-sensing regions of the hypothalamus, and/or regions lacking a complete blood brain barrier, possibly through altered pro- and anti-inflammatory signaling in these regions. PMID:24291211

Radler, Morgan E; Hale, Matthew W; Kent, Stephen

2014-05-01

48

Stiffness-Activated GEF-H1 Expression Exacerbates LPS-Induced Lung Inflammation  

PubMed Central

Acute lung injury (ALI) is accompanied by decreased lung compliance. However, a role of tissue mechanics in modulation of inflammation remains unclear. We hypothesized that bacterial lipopolysacharide (LPS) stimulates extracellular matrix (ECM) production and vascular stiffening leading to stiffness-dependent exacerbation of endothelial cell (EC) inflammatory activation and lung barrier dysfunction. Expression of GEF-H1, ICAM-1, VCAM-1, ECM proteins fibronectin and collagen, lysyl oxidase (LOX) activity, interleukin-8 and activation of Rho signaling were analyzed in lung samples and pulmonary EC grown on soft (1.5 or 2.8 kPa) and stiff (40 kPa) substrates. LPS induced EC inflammatory activation accompanied by expression of ECM proteins, increase in LOX activity, and activation of Rho signaling. These effects were augmented in EC grown on stiff substrate. Stiffness-dependent enhancement of inflammation was associated with increased expression of Rho activator, GEF-H1. Inhibition of ECM crosslinking and stiffening by LOX suppression reduced EC inflammatory activation and GEF-H1 expression in response to LPS. In vivo, LOX inhibition attenuated LPS-induced expression of GEF-H1 and lung dysfunction. These findings present a novel mechanism of stiffness-dependent exacerbation of vascular inflammation and escalation of ALI via stimulation of GEF-H1 - Rho pathway. This pathway represents a fundamental mechanism of positive feedback regulation of inflammation. PMID:24739883

Mambetsariev, Isa; Tian, Yufeng; Wu, Tinghuai; Lavoie, Tera; Solway, Julian; Birukov, Konstantin G.; Birukova, Anna A.

2014-01-01

49

Yohimbine promotes cardiac NE release and prevents LPS-induced cardiac dysfunction via blockade of presynaptic ?2A-adrenergic receptor.  

PubMed

Myocardial depression is an important contributor to mortality in sepsis. We have recently demonstrated that ?2-adrenoceptor (AR) antagonist, yohimbine (YHB), attenuates lipopolysaccharide (LPS)-induced myocardial depression. However, the mechanisms for this action of YHB are unclear. Here, we demonstrated that YHB decreased nitric oxide (NO) and tumor necrosis factor-alpha (TNF-?) levels in the myocardium and plasma, attenuated cardiac and hepatic dysfunction, but not kidney and lung injuries in endotoxemic mice. Immunohistochemical analysis revealed that cardiac ?2A-AR was mostly located in sympathetic nerve presynaptic membrane; YHB decreased cardiac ?2A-AR level and promoted cardiac norepinephrine (NE) release in endotoxemic mice. Reserpine that exhausted cardiac NE without markedly decreasing plasma NE level abrogated the inhibitory effects of YHB on cardiac TNF-? and iNOS expression as well as cardiac dysfunction, but not the suppressive effects of YHB on plasma TNF-? and NO elevation in LPS-challenged mice. Furthermore, both reserpine and YHB significantly inhibited LPS-induced myocardial apoptosis. ?1-AR, ?2-AR, but not ?1-AR antagonists reversed the inhibitory effect of YHB on LPS-stimulated myocardial apoptosis. However, ?1-AR antagonist attenuated LPS-caused cardiomyocyte apoptosis, partly abolished the protective effect of YHB on the left ventricular ejection fraction in endotoxemic mice. Altogether, these findings indicate that YHB attenuates LPS-induced cardiac dysfunction, at least in part, through blocking presynaptic ?2A-AR and thus increasing cardiac NE release. YHB-elevated cardiac NE improves cardiac function via suppressing cardiac iNOS and TNF-? expression, activating ?1-AR and inhibiting cardiomyocyte apoptosis through ?1- and ?2-AR in endotoxemic mice. However, cardiac ?1-AR activation promotes LPS-induced cardiomyocyte apoptosis. PMID:23691077

Wang, Yiyang; Yu, Xiaohui; Wang, Faqiang; Wang, Yuan; Wang, Yanping; Li, Hongmei; Lv, Xiuxiu; Lu, Daxiang; Wang, Huadong

2013-01-01

50

SeMet Mediates Anti-inflammation in LPS-Induced U937 Cells Targeting NF-?B Signaling Pathway.  

PubMed

In previous studies, selenium (Se) was reported to play critical roles in anti-inflammatory activities. Nevertheless, limited information could be obtained during inflammation about selenomethionine (SeMet) in U937 human macrophage cells. The purpose of this study was to investigate the effects of SeMet on the inflammatory responses to lipopolysaccharide (LPS)-induced U937 macrophage cells and the signaling pathways targeted. U937 cells were pretreated with SeMet (1 ?M) and subsequently induced with LPS (1 ?g/ml) for 24 h. In the cell counting kit-8 assay (CCK-8), SeMet significantly inhibits the proliferation of U937 cells. SeMet also inhibited the production of nitric oxide (NO) and prostaglandin E2 (PGE2) stimulated by LPS. In the Western blot assay and real-time polymerase chain reaction (RT-PCR), SeMet significantly reduced protein expression and production of inducible NO synthase (iNOS), tumor necrosis factor-alpha (TNF-?), and COX-2 in U937 cells. Furthermore, SeMet markedly suppressed the LPS-mediated activation of nuclear factor-kappa B (NF-?B) by blocking the degradation of inhibitor-?B proteins (I?B?) and lessening the translocations of P50 subunit content of NF-?B in the nucleus. These findings suggested the anti-inflammatory activity of SeMet in U937 cells; indicating that SeMet might be a potential treatment for inflammation therapy. PMID:25145772

Shen, Yue; Yang, Shizhou; Shi, Zhongli; Lin, Tiao; Zhu, Hanxiao; Bi, Fanggang; Liu, An; Ying, Xiaozhou; Liu, Haixiao; Yu, Kehe; Yan, Shigui

2015-04-01

51

IGF-1 attenuates LPS induced pro-inflammatory cytokines expression in buffalo (Bubalus bubalis) granulosa cells.  

PubMed

Interaction between immune and endocrine system is a diverse process influencing cellular function and homeostasis in animals. Negative energy balance (NEB) during postpartum period in dairy animals usually suppresses these systems resulting in reproductive tract infection and infertility. These negative effects could be due to competition among endocrine and immune signaling pathways for common signaling molecules. The present work studied the effect of IGF-1 (50 ng/ml) on LPS (1 ?g/ml) mediated pro-inflammatory cytokine expression (IL-1?, TNF-?, IL-6) and aromatase (CYP19A1) genes' expressions as well as proliferation of buffalo granulosa cells. The crosstalk between LPS and IGF-1 was also demonstrated through studying the activities of downstream signaling molecules (ERK1/2, Akt, NF-?B) by western blot and immunostaining. Gene expression analysis showed that IGF-1 significantly reduced the LPS induced expression of IL-1?, TNF-? and IL-6. LPS alone inhibited the CYP19A1 expression. However, co-treatment with IGF-1 reversed the inhibitory effect of LPS on CYP19A1 expression. LPS alone did not affect granulosa cell proliferation, but co-treatment with IGF-1, and IGF-1 alone enhanced the proliferation. Western blot results demonstrated that LPS caused the nuclear translocation of the NF-?B and increased the phosphorylation of ERK1/2 and Akt maximum at 15 min and 60 min, respectively. Nonetheless, co-treatment with IGF-1 delayed LPS induced phosphorylation of ERK1/2 (peak at 120 min), while promoting early Akt phosphorylation (peak at 5 min) with no effect on NF-?B translocation. Overall, IGF-1 delayed and reversed the effects of LPS, suggesting that high IGF-1 levels may combat infection during critical periods like NEB in postpartum dairy animals. PMID:25433435

Onnureddy, K; Ravinder; Onteru, Suneel Kumar; Singh, Dheer

2015-03-01

52

IL-8 signaling does not mediate intra-amniotic LPS-induced inflammation and maturation in preterm fetal lamb lung.  

PubMed

Preterm infants exposed to chorioamnionitis and preterm sheep fetuses exposed to intra-amniotic (IA) LPS have lung inflammation, increased IL-8 levels, and lung maturation. We tested the hypothesis that IL-8 signaling mediates IA LPS-induced lung inflammation and lung maturation. Two strategies were used: 1) we tested if IA injection of recombinant sheep IL-8 (rsIL-8) induced fetal inflammation and 2) if IL-8 signaling was blocked by a novel CXCR2 receptor blocker, nicotinanilide thioglycolate methyl ester (NTME). To test effects of IL-8 in the fetus, rsIL-8 was given intravascularly (50 microg) at 124 +/- 1 day of gestation (term = 150 days). A separate group of sheep was given IA rsIL-8 (100 microg) and delivered 5 h to 7 days later at 124 +/- 1 day of gestation. After confirming efficacy of the CXCR2 inhibitor, effects of IL-8 blockade were tested by injecting fetal sheep intramuscularly with NTME (10 mg) before IA injection of Escherichia coli LPS (10 mg). Sheep fetuses were delivered 1 or 7 days after injections at 124 +/- 1 day of gestation. IA rsIL-8 induced a modest fivefold increase in bronchoalveolar lavage (BAL) monocytes and neutrophils and increased lung monocyte hydrogen peroxide generation. However, rsIL-8 did not induce lung maturation. Intravascular rsIL-8 did not change fetal cardiovascular variables, blood pH, or blood leukocyte counts. Inhibition of CXCR2 decreased IA LPS-induced increases in BAL proteins at 1 day but not at 7 days. NTME did not significantly decrease IA LPS-induced BAL leukocyte influx and lung cytokine mRNA expression. Inhibition of CXCR2 did not change IA LPS-induced lung maturation. IL-8 signaling does not mediate LPS-induced lung inflammation and lung maturation. PMID:19574422

Kallapur, Suhas G; Moss, Timothy J M; Auten, Richard L; Nitsos, Ilias; Pillow, J Jane; Kramer, Boris W; Maeda, Dean Y; Newnham, John P; Ikegami, Machiko; Jobe, Alan H

2009-09-01

53

Moringa fruit inhibits LPS-induced NO/iNOS expression through suppressing the NF-? B activation in RAW264.7 cells.  

PubMed

In this study, we evaluated the anti-inflammatory effects of moringa (Moringa oleifera Lam.), a natural biologically active substance, by determining its inhibitory effects on pro-inflammatory mediators in lipopolysaccharide (LPS)-stimulated macrophage RAW264.7 cells. Extracts from different parts of moringa (root, leaf, and fruit) reduced LPS-induced nitric oxide (NO) release in a dose-dependent manner. The moringa fruit extract most effectively inhibited LPS-induced NO production and levels of inducible nitric oxide synthase (iNOS). The moringa fruit extract also was shown to suppress the production of inflammatory cytokines including IL-1?, TNF-?, and IL-6. Furthermore, moringa fruit extract inhibited the cytoplasmic degradation of I ? B -? and the nuclear translocation of p65 proteins, resulting in lower levels of NF -? B transactivation. Collectively, the results of this study demonstrate that moringa fruit extract reduces the levels of pro-inflammatory mediators including NO , IL-1?, TNF-?, and IL-6 via the inhibition of NF -? B activation in RAW264.7 cells. These findings reveal, in part, the molecular basis underlying the anti-inflammatory properties of moringa fruit extract. PMID:24117072

Lee, Hyo-Jin; Jeong, Yun-Jeong; Lee, Tae-Sung; Park, Yoon-Yub; Chae, Whi-Gun; Chung, Il-Kyung; Chang, Hyeun-Wook; Kim, Cheorl-Ho; Choi, Yung-Hyun; Kim, Wun-Jae; Moon, Sung-Kwon; Chang, Young-Chae

2013-01-01

54

Chitosan oligosaccharides block LPS-induced O-GlcNAcylation of NF-?B and endothelial inflammatory response  

PubMed Central

It is known that chitosan oligosaccharides (COS) suppress LPS-induced vascular endothelial inflammatory response by mechanism involving NF-?B blockade. It remains unknown how COS inhibit NF-?B. We provided evidence both in cultured endothelial cells and mouse model supporting a new mechanism. Regardless of the endothelial cell types, the LPS-induced NF-?B-dependent inflammatory gene expression was suppressed by COS, which was associated with reduced NF-?B nucleus translocation. LPS enhanced O-GlcNAc modification of NF-?B/p65 and activated NF-?B pathway, which could be prevented either by siRNA knockdown of O-GlcNAc transferase (OGT) or pretreatment with COS. Inhibition of either mitogen-activated protein kinase or superoxide generation abolishes LPS-induced NF-?B O-GlcNAcylation. Consistently, aortic tissues from LPS-treated mice presented enhanced NF-?B/p65 O-GlcNAcylation in association with upregulated gene expression of inflammatory cytokines in vascular tissues; however, pre-administration of COS prevented these responses. In conclusion, COS decreased OGT-dependent O-GlcNAcylation of NF-?B and thereby attenuated LPS-induced vascular endothelial inflammatory response. PMID:24274545

Li, Yu; Liu, Hongtao; Xu, Qing-Song; Du, Yu-Guang; Xu, Jian

2013-01-01

55

PARP-1 mediates LPS-induced HMGB1 release by macrophages through regulation of HMGB1 acetylation.  

PubMed

The high-mobility group box protein 1 (HMGB1) is increasingly recognized as an important inflammatory mediator. In some cases, the release of HMGB1 is regulated by poly(ADP-ribose) polymerase-1 (PARP-1), but the mechanism is still unclear. In this study, we report that PARP-1 activation contributes to LPS-induced PARylation of HMGB1, but the PARylation of HMGB1 is insufficient to direct its migration from the nucleus to the cytoplasm; PARP-1 regulates the translocation of HMGB1 to the cytoplasm through upregulating the acetylation of HMGB1. In mouse bone marrow-derived macrophages, genetic and pharmacological inhibition of PARP-1 suppressed LPS-induced translocation and release of HMGB1. Increased PARylation was accompanied with the nucleus-to-cytoplasm translocation and release of HMGB1 upon LPS exposure, but PARylated HMGB1 was located at the nucleus, unlike acetylated HMGB1 localized at the cytoplasm in an import assay. PARP inhibitor and PARP-1 depletion decreased the activity ratio of histone acetyltransferases to histone deacetylases that elevated after LPS stimulation and impaired LPS-induced acetylation of HMGB1. In addition, PARylation of HMGB1 facilitates its acetylation in an in vitro enzymatic reaction. Furthermore, reactive oxygen species scavenger (N-acetyl-l-cysteine) and the ERK inhibitor (FR180204) impaired LPS-induced PARP activation and HMGB1 release. Our findings suggest that PARP-1 regulates LPS-induced acetylation of HMGB1 in two ways: PARylating HMGB1 to facilitate the latter acetylation and increasing the activity ratio of histone acetyltransferases to histone deacetylases. These studies revealed a new mechanism of PARP-1 in regulating the inflammatory response to endotoxin. PMID:25392528

Yang, Zhiyong; Li, Li; Chen, Lijuan; Yuan, Weiwei; Dong, Liming; Zhang, Yushun; Wu, Heshui; Wang, Chunyou

2014-12-15

56

Anti-inflammatory Effects of Cavidine In Vitro and In Vivo, a Selective COX-2 Inhibitor in LPS-Induced Peritoneal Macrophages of Mouse.  

PubMed

Cavidine is an isoquinoline alkaloid which is isolated from Corydalis impatiens. In traditional Tibetan herb, C. impatiens has been widely used for treatment of skin injuries, hepatitis, cholecystitis, and scabies. The present study aimed to evaluate its anti-inflammatory effect and investigate the mechanisms underlying this anti-inflammatory action. We used different inflammation model animals and lipopolysaccharide (LPS)-induced murine peritoneal macrophages to examine the anti-inflammatory function of cavidine. Results indicated pretreatment with cavidine (i.p.) decreased xylene-induced ear edema, formaldehyde-induced paw edema, leukocyte number, and the level of nitric oxide (NO), prostaglandin E2 (PGE2), and tumor necrosis factor-alpha (TNF-?) in acetic acid-induced peritonitis in mice. The data also demonstrated that cavidine significantly inhibited LPS-induced TNF-?, interleukin-6 (IL-6), and NO production in peritoneal macrophages. Moreover, cavidine regulated the expression of cyclooxygenase-2 (COX-2) instead of cyclooxygenase-1 (COX-1) at protein levels. These results suggested that cavidine is a selective COX-2 inhibitor which possesses an anti-inflammatory activity. PMID:25373916

Niu, Xiaofeng; Zhang, Hailin; Li, Weifeng; Mu, Qingli; Yao, Huan; Wang, Yu

2015-04-01

57

p52-independent nuclear translocation of RelB promotes LPS-induced attachment  

SciTech Connect

The NF-{kappa}B signaling pathways have a critical role in the development and progression of various cancers. In this study, we demonstrated that the small cell lung cancer cell line (SCLC) H69 expressed a unique NF-{kappa}B profile as compared to other cancer cell lines. The p105/p50, p100/p52, c-Rel, and RelB protein and mRNA transcripts were absent in H69 cells but these cells expressed RelA/p65. The activation of H69 cells by lipopolysaccharide (LPS) resulted in the induction of RelB and p100 expression. The treatment also induced the nuclear translocation of RelB without the processing of p100 to p52. Furthermore, LPS-induced {beta}1 integrin expression and cellular attachment through an NF-{kappa}B-dependent mechanism. Blocking RelB expression prevented the increase in the expression of {beta}1 integrin and the attachment of H69. Taken together, the results suggest that RelB was responsible for the LPS-mediated attachment and may play an important role in the progression of some cancers.

Saito, T. [Laboratory of Immunology, National Institute on Aging, NIH Biomedical Research Center, Baltimore, MD 21224 (United States)] [Laboratory of Immunology, National Institute on Aging, NIH Biomedical Research Center, Baltimore, MD 21224 (United States); Sasaki, C.Y., E-mail: sasakic@grc.nia.nih.gov [Laboratory of Immunology, National Institute on Aging, NIH Biomedical Research Center, Baltimore, MD 21224 (United States); Rezanka, L.J.; Ghosh, P.; Longo, D.L. [Laboratory of Immunology, National Institute on Aging, NIH Biomedical Research Center, Baltimore, MD 21224 (United States)] [Laboratory of Immunology, National Institute on Aging, NIH Biomedical Research Center, Baltimore, MD 21224 (United States)

2010-01-01

58

Herbal medicine IMOD suppresses LPS-induced production of proinflammatory cytokines in human dendritic cells  

PubMed Central

Traditional medicines that stimulate or modulate the immune system can be used as innovative approaches to treat immunological diseases. The herbal medicine IMOD has been shown to strongly modulate immune responses in several animal studies as well as in clinical trials. However, little is known about the mechanisms of IMOD to modulate immunity. Here we have investigated whether IMOD modulates the immunological function of human dendritic cells (DCs). IMOD alone did not induce DC maturation nor production of cytokines. Notably, IMOD decreased the production of pro-inflammatory cytokines IL-6, IL-12 p70, and TNF? by LPS-activated DCs at both mRNA and protein levels in a dose dependent manner. In contrast, treatment with IMOD did not affect LPS induced-production of the anti-inflammatory cytokine IL-10. Furthermore, IMOD inhibited T cell activation/proliferation by LPS-treated DCs and skewed T-cells responses toward the T helper type 2 polarization. These data strongly indicate that IMOD has a potent immunomodulatory ability that affects TLR signaling and thereby modulates DC function. Insight into the immunomodulatory effect of herbal medicine IMOD may provide innovative strategies to affect the immune system and to help combat various diseases.

Mirzaee, Saeedeh; Drewniak, Agata; Sarrami-Forooshani, Ramin; Kaptein, Tanja M.; Gharibdoost, Farhad; Geijtenbeek, Teunis B. H.

2015-01-01

59

Granzyme K synergistically potentiates LPS-induced cytokine responses in human monocytes  

PubMed Central

Granzymes are serine proteases released by cytotoxic lymphocytes to induce apoptosis in virus-infected cells and tumor cells. Evidence is emerging that granzymes also play a role in controlling inflammation. Granzyme serum levels are elevated in patients with autoimmune diseases and infections, including sepsis. However, the function of extracellular granzymes in inflammation largely remains unknown. Here, we show that granzyme K (GrK) binds to Gram-negative bacteria and their cell-wall component lipopolysaccharide (LPS). GrK synergistically enhances LPS-induced cytokine release in vitro from primary human monocytes and in vivo in a mouse model of LPS challenge. Intriguingly, these extracellular effects are independent of GrK catalytic activity. GrK disaggregates LPS from micelles and augments LPS–CD14 complex formation, thereby likely boosting monocyte activation by LPS. We conclude that extracellular GrK is an unexpected direct modulator of LPS–TLR4 signaling during the antimicrobial innate immune response. PMID:24711407

Wensink, Annette C.; Kemp, Vera; Fermie, Job; García Laorden, M. Isabel; van der Poll, Tom; Hack, C. Erik; Bovenschen, Niels

2014-01-01

60

Granzyme K synergistically potentiates LPS-induced cytokine responses in human monocytes.  

PubMed

Granzymes are serine proteases released by cytotoxic lymphocytes to induce apoptosis in virus-infected cells and tumor cells. Evidence is emerging that granzymes also play a role in controlling inflammation. Granzyme serum levels are elevated in patients with autoimmune diseases and infections, including sepsis. However, the function of extracellular granzymes in inflammation largely remains unknown. Here, we show that granzyme K (GrK) binds to Gram-negative bacteria and their cell-wall component lipopolysaccharide (LPS). GrK synergistically enhances LPS-induced cytokine release in vitro from primary human monocytes and in vivo in a mouse model of LPS challenge. Intriguingly, these extracellular effects are independent of GrK catalytic activity. GrK disaggregates LPS from micelles and augments LPS-CD14 complex formation, thereby likely boosting monocyte activation by LPS. We conclude that extracellular GrK is an unexpected direct modulator of LPS-TLR4 signaling during the antimicrobial innate immune response. PMID:24711407

Wensink, Annette C; Kemp, Vera; Fermie, Job; García Laorden, M Isabel; van der Poll, Tom; Hack, C Erik; Bovenschen, Niels

2014-04-22

61

Simvastatin inhibits LPS-induced alveolar bone loss during metabolic syndrome.  

PubMed

Studies in recent years have shown a positive relationship between metabolic syndrome (MS) and periodontal disease (PD). Given that patients with MS take statins to reduce cholesterol, and statins also have anti-inflammatory effects, it is important to determine if statin intake hinders the progression of MS-associated PD. In this study, PD was induced in Zucker fat rats (ZFRs), an animal model for MS, and in control lean rats by periodontal injection of Aggregatibacter actinomycetemcomitans lipopolysaccharide (LPS), while simvastatin was given to some of the rats via gavage. After 4 wk of treatment, alveolar bone loss was determined by micro-computed tomography. To explore the underlying mechanisms, we determined the effect of simvastatin on tissue inflammation and the expression of molecules involved in osteoclastogenesis. Results showed that while bone loss was increased by LPS in both ZFRs and the control lean rats, it was significantly more in the former than the latter. Simvastatin effectively alleviated bone loss in both ZFRs and the control rats. Results also showed that LPS stimulated leukocyte tissue infiltration and expression of molecules for osteoclastogenesis, but simvastatin significantly modulated the stimulation. This study demonstrated that simvastatin inhibited LPS-induced alveolar bone loss and periodontal tissue inflammation in rats with MS. PMID:24352501

Jin, J; Machado, E R; Yu, H; Zhang, X; Lu, Z; Li, Y; Lopes-Virella, M F; Kirkwood, K L; Huang, Y

2014-03-01

62

Protective Effects of Baicalin on Decidua Cells of LPS-Induced Mice Abortion  

PubMed Central

The study was carried out to investigate the protective effects of Baicalin on decidual cells of LPS-induced abortion mice. In the in vitro experiment, the decidual cells were cultured by uterus tissue mass cultivation sampled at day 6 of pregnancy, and gradient concentrations of LPS were used to determine the optimal LPS concentration of the injured decidual cells model. The injured decidual cells were treated with Baicalin (4??g/mL) to determine the protective role of Baicalin. In the in vivo experiment, lipopolysaccharide (LPS) was injected intravenously via the tail vein to induce abortion at day 6 of pregnancy, and the mice were given different concentrations of Baicalin by oral gavage consecutively at days 7 to 8 of pregnancy. On day 9 of gestation, the mice were sacrificed. The TNF and progesterone contents in the serum were assayed by ELISA. The results clearly revealed that Baicalin can prevent the injury to decidual cells from LPS dose dependently, TNF was decreased significantly (P < 0.01) compared to LPS group, and there was no effect on the progesterone. These findings suggest that Baicalin has protective effects on the injured decidual cells in the pregnant mice. PMID:25386564

Wang, Xiaodan; Zhao, Yantao; Zhong, Xiuhui

2014-01-01

63

Involvement of mitogen-activated protein kinases and NF{kappa}B in LPS-induced CD40 expression on human monocytic cells  

SciTech Connect

CD40 is a costimulatory molecule linking innate and adaptive immune responses to bacterial stimuli, as well as a critical regulator of functions of other costimulatory molecules. The mechanisms regulating lipopolysaccharide (LPS)-induced CD40 expression have not been adequately characterized in human monocytic cells. In this study we used a human monocytic cell line, THP-1, to investigate the possible mechanisms of CD40 expression following LPS exposure. Exposure to LPS resulted in a dose- and time-dependent increase in CD40 expression. Further studies using immunoblotting and pharmacological inhibitors revealed that mitogen-activated protein kinases (MAPKs) and NF{kappa}B were activated by LPS exposure and involved in LPS-induced CD40 expression. Activation of MAPKs was not responsible for LPS-induced NF{kappa}B activation. TLR4 was expressed on THP-1 cells and pretreatment of cells with a Toll-like receptor 4 (TLR4) neutralizing antibody (HTA125) significantly blunted LPS-induced MAPK and NF{kappa}B activation and ensuing CD40 expression. Additional studies with murine macrophages expressing wild type and mutated TLR4 showed that TLR4 was implicated in LPS-induced ERK and NF{kappa}B activation, and CD40 expression. Moreover, blockage of MAPK and NF{kappa}B activation inhibited LPS-induced TLR4 expression. In summary, LPS-induced CD40 expression in monocytic cells involves MAPKs and NF{kappa}B.

Wu Weidong [Center for Environmental Medicine, Asthma, and Lung Biology, University of North Carolina, Chapel Hill, North Carolina 27599 (United States)]|[Department of Pediatrics, University of North Carolina, Chapel Hill, North Carolina 27599 (United States)], E-mail: Weidong_Wu@med.unc.edu; Alexis, Neil E. [Center for Environmental Medicine, Asthma, and Lung Biology, University of North Carolina, Chapel Hill, North Carolina 27599 (United States)]|[Department of Pediatrics, University of North Carolina, Chapel Hill, North Carolina 27599 (United States); Chen Xian [Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, North Carolina 27599 (United States)]|[Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, North Carolina 27599 (United States); Bromberg, Philip A. [Center for Environmental Medicine, Asthma, and Lung Biology, University of North Carolina, Chapel Hill, North Carolina 27599 (United States)]|[Department of Medicine, University of North Carolina, Chapel Hill, North Carolina 27599 (United States); Peden, David B. [Center for Environmental Medicine, Asthma, and Lung Biology, University of North Carolina, Chapel Hill, North Carolina 27599 (United States)]|[Department of Pediatrics, University of North Carolina, Chapel Hill, North Carolina 27599 (United States)]|[Department of Medicine, University of North Carolina, Chapel Hill, North Carolina 27599 (United States)

2008-04-15

64

Pulmonary and Systemic Expression of Monocyte Chemotactic Proteins in Preterm Sheep Fetuses Exposed to LPS Induced Chorioamnionitis  

PubMed Central

Monocyte chemoattractant proteins (MCP-1 and MCP-2) mediate monocyte and T-lymphocyte chemotaxis, and IL-1 contributes to the pathogenesis of chorioamnionitis-induced lung inflammation and fetal inflammatory responses. We tested the hypothesis that IL-1 mediates the systemic and pulmonary induction of MCP-1 and MCP-2 in response to lipopolysaccharide (LPS) induced chorioamnionitis. MCP-1 mRNA, MCP-2 mRNA and MCP-1 protein expression were measured in two models: 1) intra-amniotic LPS and 2) intra-amniotic recombinant sheep IL-1? given at varying intervals prior to preterm delivery at 124d gestational age. Intra-amniotic LPS or IL-1? induced MCP-1 mRNA and protein and MCP-2 mRNA in fetal lung many fold at 1-2d. LPS induced intense MCP-1 expression in sub-epithelial mesenchymal cells and interstitial inflammatory cells in the lung. Inhibition of IL-1 signaling with recombinant human IL-1 receptor antagonist (rhIL-1ra) did not attenuate LPS induced increase in MCP-1 or MCP-2 expression. MCP-1 and MCP-2 were not induced in liver or chorioamnion, but MCP-1 increased in cord plasma. LPS or IL-1 can induce robust expression of MCP-1 or MCP-2 in the fetal lung. LPS induction of MCP-1 is not IL-1 dependent in fetal sheep. MCP-1 and MCP-2 may be significant contributors to fetal inflammation. PMID:20703142

Shah, Tushar A.; Hillman, Noah H.; Nitsos, Ilias; Polglase, Graeme R.; Pillow, J. Jane; Newnham, John P.; Jobe, Alan H.; Kallapur, Suhas G.

2011-01-01

65

Protective effect of sanguinarine on LPS-induced endotoxic shock in mice and its effect on LPS-induced COX-2 expression and COX-2 associated PGE2 release from peritoneal macrophages.  

PubMed

The quaternary ammonium salt, sanguinarine (SG) was reported to possess widespread anti-microbial and anti-inflammatory effects in experimental animals and it has been used to treat many inflammatory diseases. The aim of this study was to evaluate the anti-inflammatory effect and the possible mechanisms underlying the anti-inflammatory activity of SG. Experimentally-induced mice ES model and LPS-induced peritoneal macrophages were used to examine the anti-inflammatory function of SG. In this study, SG pretreatment significantly increased the survival rate of mice from 25% to 58%, 75% and 91% respectively. The production of PGE2 in BALF, the lung MPO activity and the (W/D) weight ratios were also markedly reduced. In addition, immunohistochemical analysis showed that the expression of COX-2 was significantly suppressed in vivo. We also evaluated the effect of SG in LPS-induced peritoneal macrophages to clarify the possible mechanism. The data indicated that SG greatly inhibited the production of PGE2, and it also decreased COX-2 protein expression, without affecting COX-1 expression, in LPS-stimulated peritoneal macrophages. Taken all together, SG potently protected against LPS-induced ES, and our results suggest that the possible mechanism may be relevant to COX-2 regulation. PMID:25063710

Li, Weifeng; Li, Huani; Mu, Qingli; Zhang, Hailin; Yao, Huan; Li, Jiaoshe; Niu, Xiaofeng

2014-10-01

66

Impeding the interaction between Nur77 and p38 reduces LPS-induced inflammation.  

PubMed

Sepsis, a hyperinflammatory response that can result in multiple organ dysfunctions, is a leading cause of mortality from infection. Here, we show that orphan nuclear receptor Nur77 (also known as TR3) can enhance resistance to lipopolysaccharide (LPS)-induced sepsis in mice by inhibiting NF-?B activity and suppressing aberrant cytokine production. Nur77 directly associates with p65 to block its binding to the ?B element. However, this function of Nur77 is countered by the LPS-activated p38? phosphorylation of Nur77. Dampening the interaction between Nur77 and p38? would favor Nur77 suppression of the hyperinflammatory response. A compound, n-pentyl 2-[3,5-dihydroxy-2-(1-nonanoyl) phenyl]acetate, screened from a Nur77-biased library, blocked the Nur77-p38? interaction by targeting the ligand-binding domain of Nur77 and restored the suppression of the hyperinflammatory response through Nur77 inhibition of NF-?B. This study associates the nuclear receptor with immune homeostasis and implicates a new therapeutic strategy to treat hyperinflammatory responses by targeting a p38? substrate to modulate p38?-regulated functions. PMID:25822914

Li, Li; Liu, Yuan; Chen, Hang-Zi; Li, Feng-Wei; Wu, Jian-Feng; Zhang, Hong-Kui; He, Jian-Ping; Xing, Yong-Zhen; Chen, Yan; Wang, Wei-Jia; Tian, Xu-Yang; Li, An-Zhong; Zhang, Qian; Huang, Pei-Qiang; Han, Jiahuai; Lin, Tianwei; Wu, Qiao

2015-05-01

67

Exogenous rhTRX reduces lipid accumulation under LPS-induced inflammation  

PubMed Central

Redox-regulating molecule, recombinant human thioredoxin (rhTRX) which shows anti-inflammatory, and anti-oxidative effects against lipopolysaccharide (LPS)-stimulated inflammation and regulate protein expression levels. LPS-induced reactive oxygen intermediates (ROI) and NO production were inhibited by exogenous rhTRX. We identified up/downregulated intracellular proteins under the LPS-treated condition in exogenous rhTRX-treated A375 cells compared with non-LPS-treated cells via 2-DE proteomic analysis. Also, we quantitatively measured cytokines of in vivo mouse inflammation models using cytometry bead array. Exogenous rhTRX inhibited LPS-stimulated production of ROI and NO levels. TIP47 and ATP synthase may influence the inflammation-related lipid accumulation by affecting lipid metabolism. The modulation of skin redox environments during inflammation is most likely to prevent alterations in lipid metabolism through upregulation of TIP47 and ATP synthase and downregulation of inflammatory cytokines. Our results demonstrate that exogenous rhTRX has anti-inflammatory properties and intracellular regulatory activity in vivo and in vitro. Monitoring of LPS-stimulated pro-inflammatory conditions treated with rhTRX in A375 cells could be useful for diagnosis and follow-up of inflammation reduction related with candidate proteins. These results have a therapeutic role in skin inflammation therapy. PMID:24406320

Han, Gi-Yeon; Lee, Eun-Kyung; Park, Hey-won; Kim, Hyun-Jung; Kim, Chan-Wha

2014-01-01

68

Eukaryotic elongation factor 2 controls TNF-? translation in LPS-induced hepatitis  

PubMed Central

Bacterial LPS (endotoxin) has been implicated in the pathogenesis of acute liver disease through its induction of the proinflammatory cytokine TNF-?. TNF-? is a key determinant of the outcome in a well-established mouse model of acute liver failure during septic shock. One possible mechanism for regulating TNF-? expression is through the control of protein elongation during translation, which would allow rapid cell adaptation to physiological changes. However, the regulation of translational elongation is poorly understood. We found that expression of p38?/? MAPK proteins is required for the elongation of nascent TNF-? protein in macrophages. The MKK3/6-p38?/? pathway mediated an inhibitory phosphorylation of eukaryotic elongation factor 2 (eEF2) kinase, which in turn promoted eEF2 activation (dephosphorylation) and subsequent TNF-? elongation. These results identify a new signaling pathway that regulates TNF-? production in LPS-induced liver damage and suggest potential cell-specific therapeutic targets for liver diseases in which TNF-? production is involved. PMID:23202732

González-Terán, Bárbara; Cortés, José R.; Manieri, Elisa; Matesanz, Nuria; Verdugo, ?ngeles; Rodríguez, María E.; González-Rodríguez, ?gueda; Valverde, ?ngela; Martín, Pilar; Davis, Roger J.; Sabio, Guadalupe

2012-01-01

69

Escherichia coli LPS induces heat shock protein 25 in intestinal epithelial cells through MAP kinase activation.  

PubMed

Protection of colonic epithelial integrity and function is critical, because compromises in mucosal functions can lead to adverse and potentially life-threatening effects. The gut flora may contribute to this protection, in part, through the sustained induction of cytoprotective heat shock proteins (HSPs) in surface colonocytes. In this study, we investigated whether Escherichia coli LPS mediates bacteria-induced HSP by using cultured young adult mouse colon (YAMC) cells, an in vitro model of the colonic epithelium. E. coli LPS led to an epithelial cell-type specific induction of HSP25 in a time- and concentration-dependent manner, an effect that did not involve changes in HSP72. YAMC cells expressed the toll-like receptors (TLR)2 and TLR4 but not the costimulatory CD14 molecule. Whereas LPS stimulated both the p38 and ERK1/2 but not the stress-activated protein kinase/c-Jun NH(2)-terminal kinase, signaling pathways in the YAMC cells, all three were stimulated in RAW macrophage cells (in which no LPS-induced HSP25 expression was observed). The p38 inhibitor SB-203580 and the MAP kinase kinase-1 inhibitor PD-98059 inhibited HSP25 induction by LPS. LPS treatment also conferred protection against actin depolymerization induced by the oxidant monochloramine. The HSP25 dependence of the LPS protective effect was outlined in inhibitor studies and through adenovirus-mediated overexpression of HSP25. In conclusion, LPS may be an important mediator of enteric bacteria-induced expression of intestinal epithelial HSP25, an effect that may contribute to filamentous actin stabilization under physiological as well as pathophysiological conditions and thus protection of colonic epithelial integrity. PMID:14630641

Kojima, Keishi; Musch, Mark W; Ropeleski, Mark J; Boone, David L; Ma, Averil; Chang, Eugene B

2004-04-01

70

Myeloid depletion of SOCS3 enhances LPS-induced acute lung injury through CCAAT/enhancer binding protein ? pathway  

PubMed Central

Although uncontrolled inflammatory response plays a central role in the pathogenesis of acute lung injury (ALI), the precise molecular mechanisms underlying the development of this disorder remain poorly understood. SOCS3 is an important negative regulator of IL-6-type cytokine signaling. SOCS3 is induced in lung during LPS-induced lung injury, suggesting that generation of SOCS3 may represent a regulatory product during ALI. In the current study, we created mice lacking SOCS3 expression in macrophages and neutrophils (LysM-cre SOCS3fl/fl). We evaluated the lung inflammatory response to LPS in both LysM-cre SOCS3fl/fl mice and the wild-type (WT) mice (SOCS3fl/fl). LysM-cre SOCS3fl/fl mice displayed significant increase of the lung permeability index (lung vascular leak of albumin), neutrophils, lung neutrophil accumulation (myeloperoxidase activity), and proinflammatory cytokines/chemokines in bronchial alveolar lavage fluids compared to WT mice. These phenotypes were consistent with morphological evaluation of lung, which showed enhanced inflammatory cell influx and intra-alveolar hemorrhage. We further identify the transcription factor, CCAAT/enhancer-binding protein (C/EBP) ? as a critical downstream target of SOCS3 in LPS-induced ALI. These results indicate that SOCS3 has a protective role in LPS-induced ALI by suppressing C/EBP? activity in the lung. Elucidating the function of SOCS3 would represent prospective targets for a new generation of drugs needed to treat ALI.—Yan, C., Ward, P. A., Wang, X., Gao, H. Myeloid depletion of SOCS3 enhances LPS-induced acute lung injury through CCAAT/enhancer binding protein ? pathway. PMID:23585399

Yan, Chunguang; Ward, Peter A.; Wang, Ximo; Gao, Hongwei

2013-01-01

71

Wogonin inhibits LPS-induced tumor angiogenesis via suppressing PI3K/Akt/NF-?B signaling.  

PubMed

Wogonin has been shown to have anti-angiogenesis and anti-tumor effects. However, whether wogonin inhibits LPS-induced tumor angiogenesis is not well known. In this study, we investigated the effect of wogonin on inhibiting LPS-induced tumor angiogenesis and further probed the underlying mechanisms. ELISA results revealed that wogonin could suppress LPS-induced VEGF secretion from tumor cells. Transwell assay, tube formation assay, rat aortic ring assay and CAM model were used to evaluate the effect of wogonin on angiogenesis induced by MCF-7 cell (treated with LPS) in vitro and in vivo. The inhibitory effect of wogonin on angiogenesis in LPS-treated MCF-7 cells was then confirmed by the above in vitro and in vivo assays. The study of the molecular mechanism showed that wogonin could suppress PI3K/Akt signaling activation. Moreover, wogonin inhibited nuclear translocation of NF-?B and its binding to DNA. The result of real-time PCR and luciferase reporter assay suggested that VEGF expression was down-regulated by wogonin primarily at the transcriptional level. IGF-1 and p65 expression plasmid were used to activate PI3K/Akt and NF-?B pathways, and to observe the effect of wogonin on the simualtion of PI3K/Akt/NF-?B signaling. Taken together, the result suggested that wogonin was a potent inhibitor of tumor angiogenesis and provided a new insight into the mechanisms of wogonin against cancer. PMID:24858369

Zhao, Kai; Song, Xiuming; Huang, Yujie; Yao, Jing; Zhou, Mi; Li, Zhiyu; You, Qidong; Guo, Qinglong; Lu, Na

2014-08-15

72

Tanshinone IIA therapeutically reduces LPS-induced acute lung injury by inhibiting inflammation and apoptosis in mice  

PubMed Central

Aim: To study the effects of tanshinone IIA (TIIA) on lipopolysaccharide (LPS)-induced acute lung injury in mice and the underlying mechanisms. Methods: Mice were injected with LPS (10 mg/kg, ip), then treated with TIIA (10 mg/kg, ip). Seven hours after LPS injection, the lungs were collected for histological study. Protein, LDH, TNF-? and IL-1? levels in bronchoalveolar lavage fluid (BALF) and myeloperoxidase (MPO) activity in lungs were measured. Cell apoptosis and Bcl-2, caspase-3, NF-?B and HIF-1? expression in lungs were assayed. Results: LPS caused marked histological changes in lungs, accompanied by significantly increased lung W/D ratio, protein content and LDH level in BALF, and Evans blue leakage. LPS markedly increased neutrophil infiltration in lungs and inflammatory cytokines in BALF. Furthermore, LPS induced cell apoptosis in lungs, as evidenced by increased TUNEL-positive cells, decreased Bcl-2 content and increased cleaved caspase-3 content. Moreover, LPS significantly increased the expression of NF-?B and HIF-1? in lungs. Treatment of LPS-injected mice with TIIA significantly alleviated these pathological changes in lungs. Conclusion: TIIA alleviates LPS-induced acute lung injury in mice by suppressing inflammatory responses and apoptosis, which is mediated via inhibition of the NF-?B and HIF-1? pathways. PMID:25544360

Xu, Min; Cao, Fa-le; Zhang, Yu-fei; Shan, Liang; Jiang, Xiao-ling; An, Xiao-jing; Xu, Wei; Liu, Xiu-zhi; Wang, Xiao-yan

2015-01-01

73

A20 expression in dendritic cells protects mice from LPS-induced mortality.  

PubMed

DCs contribute to immune homeostasis under physiological conditions and regulate the immune activation during infection. The deubiquitinase A20 inhibits the activation of NF-?B-dependent immune reactions, and prevents the hyperactivation of DCs under steady-state conditions. However, the role of DC-specific A20 under pathological conditions is unknown. Here, we demonstrate that upon injection of low-dose LPS, mice with DC-specific A20 deletion (CD11c-Cre A20(fl/fl) ) died within 6 h, whereas A20(fl/fl) controls survived. LPS-induced mortality in CD11c-Cre A20(fl/fl) mice was characterized by increased serum levels of IL-2, IL-10, IL-12, IFN-?, and TNF. Upon LPS stimulation, the activation of NF-?B and ERK-NFATc3 pathways were enhanced in A20-deficient DCs, resulting in an increased production of IL-2, IL-12, and TNF both in vitro and in vivo. Targeted inhibition of ERK in A20-deficient DCs abolished the increased production of IL-2. A20-deficient DCs failed to induce LPS tolerance, which was independent of T cells and the intestinal flora, since T-cell depletion and decolonization of CD11c-Cre A20(fl/fl) mice could not prevent death of LPS-challenged CD11c-Cre A20(fl/fl) mice. In conclusion, these findings show that DC-specific A20 preserves immune homeostasis in steady-state conditions and is also required for LPS tolerance. PMID:25472594

Xuan, Nguyen Thi; Wang, Xu; Nishanth, Gopala; Waisman, Ari; Borucki, Katrin; Isermann, Berend; Naumann, Michael; Deckert, Martina; Schlüter, Dirk

2015-03-01

74

Autotaxin downregulates LPS-induced microglia activation and pro-inflammatory cytokines production.  

PubMed

Inflammation is essential in defense against infection or injury. It is tightly regulated, as over-response can be detrimental, especially in immune-privileged organs such as the central nervous system (CNS). Microglia constitutes the major source of inflammatory factors, but are also involved in the regulation of the inflammation and in the reparation. Autotaxin (ATX), a phospholipase D, converts lysophosphatidylcholine (LPC) into lysophosphatidic acid (LPA) and is upregulated in several CNS injuries. LPA, a pleiotropic immunomodulatory factor, can induce multiple cellular processes including morphological changes, proliferation, death, and survival. We investigated ATX effects on microglia inflammatory response to lipopolysaccharide (LPS), mimicking gram-negative infection. Murine BV-2 microglia and stable transfected, overexpressing ATX-BV-2 (A?+) microglia were treated with LPS. Tumor necrosis factor ? (TNF?), interleukin (IL)-6, and IL-10 mRNA and proteins levels were examined by qRT-PCR and ELISA, respectively. Secreted LPA was quantified by a radioenzymatic assay and microglial activation markers (CD11b, CD14, B7.1, and B7.2) were determined by flow cytometry. ATX expression and LPA production were significantly enhanced in LPS treated BV-2 cells. LPS induction of mRNA and protein level for TNF? and IL-6 were inhibited in A+ cells, while IL-10 was increased. CD11b, CD14, and B7.1, and B7.2 expressions were reduced in A+ cells. Our results strongly suggest deactivation of microglia and an IL-10 inhibitory of ATX with LPS induced microglia activation. PMID:25053164

Awada, Rana; Saulnier-Blache, Jean Sébastien; Grès, Sandra; Bourdon, Emmanuel; Rondeau, Philippe; Parimisetty, Avinash; Orihuela, Ruben; Harry, G Jean; d'Hellencourt, Christian Lefebvre

2014-12-01

75

COX1 and COX2 contribute differentially to the LPS-induced release of PGE 2 and TxA 2 in liver macrophages  

Microsoft Academic Search

LPS induces an immediate release of thromboxane TxA2 and a delayed release of PGE2. Dexamethasone suppresses the LPS-induced release of TxA2 and PGE2. In the first 8h after LPS addition, the specific COX-2 inhibitor SC236 inhibits the PGE2 and TxA2 release by about 80% and 20%, whereas the release of PGE2 and TxA2 between 8 and 24h is inhibited by

Yevgeniya Bezugla; Angelika Kolada; Sabine Kamionka; Brigitte Bernard; Roland Scheibe; Peter Dieter

2006-01-01

76

2-phenylethynesulfonamide Prevents Induction of Pro-inflammatory Factors and Attenuates LPS-induced Liver Injury by Targeting NHE1-Hsp70 Complex in Mice  

PubMed Central

The endotoxin-mediated production of pro-inflammatory cytokines plays an important role in the pathogenesis of liver disorders. Heat shock protein (Hsp70) overexpression has established functions in lipopolysaccharide (LPS)-mediated inflammatory response. However, little is known about the role of Hsp70 activity in LPS signaling. We hypothesized that inhibition of Hsp70 substrate binding activity can ameliorate LPS-induced liver injury by decreasing induction of pro-inflammatory factors. In this study, C57/BL6 mice were injected intraperitoneally with LPS and 2-phenylethynesulfonamide (PES), an inhibitor of Hsp70 substrate binding activity. We found that i. PES prevented LPS-induced increase in serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activity, infiltration of inflammatory cells, and liver cell apoptosis; ii. PES reduced inducible nitric oxide synthase (iNOS) protein expression as well as serum nitric oxide (NO), tumor necrosis factor-? (TNF-?), and interleukin-6 (IL-6) content in LPS-stimulated mice; iii. PES reduced the mRNA level of iNOS, TNF-?, and IL-6 in LPS-stimulated liver. iiii. PES attenuated the degradation of inhibitor of ?B-? (I?B-?) as well as the phosphorylation and nuclear translocation of nuclear factor-?B (NF-?B) in LPS-stimulated liver. Similar changes in the protein expression of inflammatory markers, I?B-? degradation, and NF-?B phosphorylation and nuclear translocation were observed in RAW 264.7 cells. Further mechanistic studies revealed that PES remarkably reduced the elevation of [Ca2+]i and intracellular pH value (pHi) in LPS-stimulated RAW 264.7 cells. Furthermore, PES significantly reduced the increase in Na+/H+ exchanger 1 (NHE1) association to Hsp70 in LPS-stimulated macrophages and liver, suggesting that NHE1-Hsp70 interaction is required for the involvement of NHE1 in the inflammation response. In conclusion, inhibition of Hsp70 substrate binding activity in vivo reduces the induction of pro-inflammatory factors and prevents LPS-induced liver injury likely by disrupting NHE1-Hsp70 interaction which consequently reduces the activation of I?B-?-NF-?B pathway in liver. PMID:23805318

Huang, Chao; Wang, Jia; Chen, Zhuo; Wang, Yuzhe; Zhang, Wei

2013-01-01

77

2-phenylethynesulfonamide Prevents Induction of Pro-inflammatory Factors and Attenuates LPS-induced Liver Injury by Targeting NHE1-Hsp70 Complex in Mice.  

PubMed

The endotoxin-mediated production of pro-inflammatory cytokines plays an important role in the pathogenesis of liver disorders. Heat shock protein (Hsp70) overexpression has established functions in lipopolysaccharide (LPS)-mediated inflammatory response. However, little is known about the role of Hsp70 activity in LPS signaling. We hypothesized that inhibition of Hsp70 substrate binding activity can ameliorate LPS-induced liver injury by decreasing induction of pro-inflammatory factors. In this study, C57/BL6 mice were injected intraperitoneally with LPS and 2-phenylethynesulfonamide (PES), an inhibitor of Hsp70 substrate binding activity. We found that i. PES prevented LPS-induced increase in serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activity, infiltration of inflammatory cells, and liver cell apoptosis; ii. PES reduced inducible nitric oxide synthase (iNOS) protein expression as well as serum nitric oxide (NO), tumor necrosis factor-? (TNF-?), and interleukin-6 (IL-6) content in LPS-stimulated mice; iii. PES reduced the mRNA level of iNOS, TNF-?, and IL-6 in LPS-stimulated liver. iiii. PES attenuated the degradation of inhibitor of ?B-? (I?B-?) as well as the phosphorylation and nuclear translocation of nuclear factor-?B (NF-?B) in LPS-stimulated liver. Similar changes in the protein expression of inflammatory markers, I?B-? degradation, and NF-?B phosphorylation and nuclear translocation were observed in RAW 264.7 cells. Further mechanistic studies revealed that PES remarkably reduced the elevation of [Ca(2+)]i and intracellular pH value (pHi) in LPS-stimulated RAW 264.7 cells. Furthermore, PES significantly reduced the increase in Na(+)/H(+) exchanger 1 (NHE1) association to Hsp70 in LPS-stimulated macrophages and liver, suggesting that NHE1-Hsp70 interaction is required for the involvement of NHE1 in the inflammation response. In conclusion, inhibition of Hsp70 substrate binding activity in vivo reduces the induction of pro-inflammatory factors and prevents LPS-induced liver injury likely by disrupting NHE1-Hsp70 interaction which consequently reduces the activation of I?B-?-NF-?B pathway in liver. PMID:23805318

Huang, Chao; Wang, Jia; Chen, Zhuo; Wang, Yuzhe; Zhang, Wei

2013-01-01

78

Anti-inflammatory activity of cinnamon water extract in vivo and in vitro LPS-induced models  

PubMed Central

Background Cinnamon bark is one of the most popular herbal ingredients in traditional oriental medicine and possesses diverse pharmacological activities including anti-bacterial, anti-viral, and anti-cancer properties. The goal of this study is to investigate the in vivo and in vitro inhibitory effect of cinnamon water extract (CWE) on lipopolysaccharide (LPS)-induced tumor necrosis factor (TNF)-? and its underlying intracellular mechanisms. Methods CWE was orally administrated to mice for 6 days prior to intraperitoneal injection of LPS. Serum levels of TNF-? and interleukin (IL)-6 were determined 1 hour after LPS stimulation. Peritoneal macrophages from thioglycollate-injected mice were isolated and assayed for viability, cytokine expression and signaling molecules upon LPS stimulation. CWE was further fractioned according to molecular size, and the levels of total polyphenols and biological activities of each fraction were measured. Results The oral administration of CWE to mice significantly decreased the serum levels of TNF-? and IL-6. CWE treatment in vitro decreased the mRNA expression of TNF-?. CWE blocked the LPS-induced degradation of I?B? as well as the activation of JNK, p38 and ERK1/2. Furthermore, size-based fractionation of CWE showed that the observed inhibitory effect of CWE in vitro occurred in the fraction containing the highest level of total polyphenols. Conclusions Treatment with CWE decreased LPS-induced TNF-? in serum. In vitro inhibition of TNF-? gene by CWE may occur via the modulation of I?B? degradation and JNK, p38, and ERK1/2 activation. Our results also indicate that the observed anti-inflammatory action of CWE may originate from the presence of polyphenols. PMID:23190501

2012-01-01

79

aged black garlic exerts anti-inflammatory effects by decreasing no and proinflammatory cytokine production with less cytoxicity in LPS-stimulated raw 264.7 macrophages and LPS-induced septicemia mice.  

PubMed

In this study, the anti-inflammatory and antisepticemic activities of a water extract of aged black garlic (AGE), which is not pungent, were compared with those of raw garlic extract (RGE). The methyl thiazolyl tetrazolium (MTT) assay showed that AGE was not toxic up to 1000 ?g/mL and was at least four times less cytotoxic than RGE. AGE significantly suppressed the production of nitric oxide (NO), tumor-necrosis factor-? (TNF-?), and prostaglandin (PG)-E2 in a dose-dependent manner in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Furthermore, the inhibitory effect of AGE on LPS-induced inflammation was confirmed by downregulation of inducible NO synthase and TNF-? mRNA expression, as well as cyclooxygenase-2 protein expression. The anti-inflammatory activities of AGE were similar to those of RGE at nontoxic concentrations up to 250 ?g/mL. Signal transduction pathway studies further indicated that both garlic extracts inhibited activation of mitogen-activated protein kinase and nuclear factor-?B induced by LPS stimulation. Treatment with both AGE and RGE in an in vivo experiment of LPS-induced endotoxemia significantly reduced the level of TNF-? and interleukin-6 in serum and completely protected against LPS-induced lethal shock in C57BL/6 mice. The results suggest that AGE is a more promising nutraceutical or medicinal agent to prevent or cure inflammation-related diseases for safety aspects compared with RGE. PMID:25238199

Kim, Min Jee; Yoo, Yung Choon; Kim, Hyun Jung; Shin, Suk Kyung; Sohn, Eun Jeong; Min, A Young; Sung, Nak Yun; Kim, Mee Ree

2014-10-01

80

Whole-body deletion of LPS-induced TNF-? factor (LITAF) markedly improves experimental endotoxic shock and inflammatory arthritis.  

PubMed

LPS-induced TNF-? factor (LITAF) mediates cytokine expression in response to endotoxin challenge. Previously, we reported that macrophage-specific LITAF-deficient (macLITAF-/-) mice exposed to LPS have a delayed onset in the serum levels of proinflammatory cytokines and prolonged persistence of anti-inflammatory cytokines, but only partial protection from endotoxic shock. We postulated that greater protection might be achieved if LITAF were deleted from all LITAF-producing cells, including macrophages. Using a Cre-loxP system, we engineered a tamoxifen-induced recombination mouse [tamLITAF(i)-/-] that resulted in whole-body LITAF deficiency. Our findings demonstrate that (i) tamLITAF(i)-/- mice are more resistant to systemic Escherichia coli LPS-induced lethality than our previous macLITAF-/- mice, providing evidence that LITAF-producing cells other than LysMCre-positive cells play an important role in mediating endotoxic shock; (ii) tamLITAF(i)-/- mice show a similar pattern of cytokine expression with decreased proinflammatory and prolonged anti-inflammatory mediators compared with WT mice; and (iii) tamLITAF(i)-/- mice, compared with WT mice, display a significant reduction in bone resorption and inflammation associated with a local chronic inflammatory disease--namely, collagen antibody-induced arthritis. Our findings offer a unique model to study the role of LITAF in systemic and chronic local inflammatory processes, and pave the way for anti-LITAF therapeutic approaches for the treatment of TNF-mediated inflammatory diseases. PMID:22160695

Merrill, Jamie C; You, Jian; Constable, Cara; Leeman, Susan E; Amar, Salomon

2011-12-27

81

Emodin Ameliorates LPS-Induced Acute Lung Injury, Involving the Inactivation of NF-?B in Mice  

PubMed Central

Acute lung injury (ALI) and its severe manifestation of acute respiratory distress syndrome (ARDS) are well-known illnesses. Uncontrolled and self-amplified pulmonary inflammation lies at the center of the pathology of this disease. Emodin, the bio-active coxund of herb Radix rhizoma Rhei, shows potent anti-inflammatory properties through inactivation of nuclear factor-?B (NF-?B). The aim of this study was to evaluate the effect of emodin on lipopolysaccharide (LPS)-induced ALI in mice, and its potential bio-mechanism. In our study, BALB/c mice were stimulated with LPS to induce ALI. After 72 h of LPS stimulation, pulmonary pathological changes, lung injury scores, pulmonary edema, myeloperoxidase (MPO) activity, total cells, neutrophils, macrophages, TNF-?, IL-6 and IL-1? in bronchoalveolar lavage fluid (BALF), and MCP-1 and E-selectin expression were notably attenuated by emodin in mice. Meanwhile, our data also revealed that emodin significantly inhibited the LPS-enhanced the phosphorylation of NF-?B p65 and NF-?B p65 DNA binding activity in lung. Our data indicates that emodin potently inhibits LPS-induced pulmonary inflammation, pulmonary edema and MCP-1 and E-selectin expression, and that these effects were very likely mediated by inactivation of NF-?B in mice. These results suggest a therapeutic potential of emodin as an anti-inflammatory agent for ALI/ARDS treatment. PMID:25347274

Xiao, Min; Zhu, Tao; Zhang, Wei; Wang, Tao; Shen, Yong-Chun; Wan, Qiong-Fang; Wen, Fu-Qiang

2014-01-01

82

Lipid emulsions differentially affect LPS-induced acute monocytes inflammation: in vitro effects on membrane remodeling and cell viability.  

PubMed

The aim of this study was to assess how lipid emulsions for parenteral nutrition affect lipopolysaccharide (LPS)-induced acute monocyte inflammation in vitro. An 18 h long LPS induced human monocyte leukemia cell stimulation was performed and the cell-growth medium was supplemented with three different industrial lipid emulsions: Intralipid(®), containing long-chain triglycerides (LCT--soybean oil); Medialipid(®), containing LCT (soybean oil) and medium-chain triglycerides (MCT--coconut oil); and SMOFlipid(®), containing LCT, MCT, omega-9 and -3 (soybean, coconut, olive and fish oils). Cell viability and apoptosis were assessed by Trypan blue exclusion and flow cytometry respectively. Monocyte composition and membrane remodeling were studied using gas chromatography and NR12S staining. Microparticles released in supernatant were measured by prothrombinase assay. After LPS challenge, both cellular necrosis and apoptosis were increased (threefold and twofold respectively) and microparticle release was enhanced (sevenfold) after supplementation with Medialipid(®) compared to Intralipid(®), SMOFlipid(®) and monocytes in the standard medium. The monocytes differentially incorporated fatty acids after lipid emulsion challenge. Finally, lipid-treated cells displayed microparticles characterized by disrupted membrane lipid order, reflecting lipid remodeling of the parental cell plasma membrane. Our data suggest that lipid emulsions differentially alter cell viability, monocyte composition and thereby microparticle release. While MCT have deleterious effects, we have shown that parenteral nutrition emulsion containing LCT or LCT and MCT associated to n-3 and n-9 fatty acids have no effect on endotoxin-induced cell death and inflammation. PMID:25038627

Boisramé-Helms, Julie; Delabranche, Xavier; Klymchenko, Andrey; Drai, Jocelyne; Blond, Emilie; Zobairi, Fatiha; Mely, Yves; Hasselmann, Michel; Toti, Florence; Meziani, Ferhat

2014-11-01

83

Salidroside Reduces Cell Mobility via NF-?B and MAPK Signaling in LPS-Induced BV2 Microglial Cells  

PubMed Central

The unregulated activation of microglia following stroke results in the production of toxic factors that propagate secondary neuronal injury. Salidroside has been shown to exhibit protective effects against neuronal death induced by different insults. However, the molecular mechanisms responsible for the anti-inflammatory activity of salidroside have not been elucidated clearly in microglia. In the present study, we investigated the molecular mechanism underlying inhibiting LPS-stimulated BV2 microglial cell mobility of salidroside. The protective effect of salidroside was investigated in microglial BV2 cell, subjected to stretch injury. Moreover, transwell migration assay demonstrated that salidroside significantly reduced cell motility. Our results also indicated that salidroside suppressed LPS-induced chemokines production in a dose-dependent manner, without causing cytotoxicity in BV2 microglial cells. Moreover, salidroside suppressed LPS-induced activation of nuclear factor kappa B (NF-?B) by blocking degradation of I?B? and phosphorylation of MAPK (p38, JNK, ERK1/2), which resulted in inhibition of chemokine expression. These results suggest that salidroside possesses a potent suppressive effect on cell migration of BV2 microglia and this compound may offer substantial therapeutic potential for treatment of ischemic strokes that are accompanied by microglial activation. PMID:24864151

Hu, Haixia; Li, Zuanfang; Zhu, Xiaoqin; Lin, Ruhui; Chen, Lidian

2014-01-01

84

Saponarin from barley sprouts inhibits NF-?B and MAPK on LPS-induced RAW 264.7 cells.  

PubMed

Saponarin (SA), a natural flavonoid, is known for its antioxidant and hepatoprotective activities. SA is the predominant compound (1142.7 ± 0.9 mg per 100 g) in barley sprouts, constituting 72% of the total polyphenol content. We investigated, for the first time, the effects of SA from barley sprouts on cellular anti-inflammatory responses. In lipopolysaccharide (LPS)-induced RAW 264.7 macrophages, SA suppressed the activation of NF-?B, as evidenced by the inhibition of NF-?B DNA binding, nuclear translocation, I?B? phosphorylation, and reporter gene expression, and it downregulated the expression of the pro-inflammatory mediator IL-6. Furthermore, SA reduced the transcription of NF-?B target molecules COX2 and FLIP inhibited the phosphorylation of mitogen-activated protein kinases ERK and p38. These results suggest that SA isolated from barley sprouts exerts anti-inflammatory effects in LPS-induced RAW 264.7 macrophages via inhibition of NF-?B, ERK and p38 signaling. Thus, SA may be a promising natural anti-inflammatory agent. PMID:25238253

Seo, Kyung Hye; Park, Mi Jin; Ra, Ji-Eun; Han, Sang-Ik; Nam, Min-Hee; Kim, Jin Hyo; Lee, Jin Hwan; Seo, Woo Duck

2014-11-01

85

Loss of Protein Kinase C-? Protects against LPS-Induced Osteolysis Owing to an Intrinsic Defect in Osteoclastic Bone Resorption  

PubMed Central

Bone remodeling is intrinsically regulated by cell signaling molecules. The Protein Kinase C (PKC) family of serine/threonine kinases is involved in multiple signaling pathways including cell proliferation, differentiation, apoptosis and osteoclast biology. However, the precise involvement of individual PKC isoforms in the regulation of osteoclast formation and bone homeostasis remains unclear. Here, we identify PKC-? as the major PKC isoform expressed among all PKCs in osteoclasts; including classical PKCs (??, ?? and ??), novel PKCs (??, ??, ?? and ??) and atypical PKCs (??/? and ??). Interestingly, pharmacological inhibition and genetic ablation of PKC-? impairs osteoclastic bone resorption in vitro. Moreover, disruption of PKC-? activity protects against LPS-induced osteolysis in mice, with osteoclasts accumulating on the bone surface failing to resorb bone. Treatment with the PKC-? inhibitor Rottlerin, blocks LPS-induced bone resorption in mice. Consistently, PKC-? deficient mice exhibit increased trabeculae bone containing residual cartilage matrix, indicative of an osteoclast-rich osteopetrosis phenotype. Cultured ex vivo osteoclasts derived from PKC-? null mice exhibit decreased CTX-1 levels and MARKS phosphorylation, with enhanced formation rates. This is accompanied by elevated gene expression levels of cathepsin K and PKC ??, ?? and ??, as well as altered signaling of pERK and pcSrc416/527 upon RANKL-induction, possibly to compensate for the defects in bone resorption. Collectively, our data indicate that PKC-? is an intrinsic regulator of osteoclast formation and bone resorption and thus is a potential therapeutic target for pathological osteolysis. PMID:23951014

Khor, Ee Cheng; Abel, Tamara; Tickner, Jennifer; Chim, Shek Man; Wang, Cathy; Cheng, Taksum; Ng, Benjamin; Ng, Pei Ying; Teguh, Dian Astari; Kenny, Jacob; Yang, Xiaohong; Chen, Honghui; Nakayama, Keiichi I.; Nakayama, Keiko; Pavlos, Nathan; Zheng, Ming H.; Xu, Jiake

2013-01-01

86

A TLR4/MD2 fusion protein inhibits LPS-induced pro-inflammatory signaling in hepatic stellate cells  

SciTech Connect

Activated hepatic stellate cells (HSCs) play a key role in hepatic fibrogenesis. In injured liver they are the main extracellular matrix protein producing cell type and further perpetuate hepatic injury by secretion of pro-inflammatory mediators. Since LPS-mediated signaling through toll-like receptor 4 (TLR4) has been identified as key fibrogenic signal in HSCs we aimed to test TLR4 as potential target of therapy via ligand-binding soluble receptors. Incubation of human HSCs with a fusion protein between the extracellular domain of TLR4 and MD2 which binds LPS inhibited LPS-induced NF{kappa}B and JNK activation. TLR4/MD2 abolished LPS-induced secretion of IL-6, IL-8, MCP1, and RANTES in HSCs. In addition, TLR4/MD2 fused to human IgG-Fc neutralized LPS activity. Since TLR4 mutant mice are resistant to liver fibrosis, the TLR4/MD2 soluble receptor might represent a new therapeutic molecule for liver fibrogenesis in vivo.

Schnabl, Bernd [Department of Medicine, University of California San Diego, 9500 Gilman Drive, MC0702, La Jolla, CA 92093 (United States)], E-mail: beschnabl@ucsd.edu; Brandl, Katharina; Fink, Marina; Gross, Philipp [Department of Internal Medicine I, University of Regensburg (Germany); Taura, Kojiro [Department of Medicine, University of California San Diego, 9500 Gilman Drive, MC0702, La Jolla, CA 92093 (United States); Gaebele, Erwin; Hellerbrand, Claus; Falk, Werner [Department of Internal Medicine I, University of Regensburg (Germany)

2008-10-17

87

Adrenaline stimulates the proliferation and migration of mesenchymal stem cells towards the LPS-induced lung injury  

PubMed Central

Bone marrow-derived mesenchymal stem cells (BMSCs) could modulate inflammation in experimental lung injury. On the other hand, adrenergic receptor agonists could increase DNA synthesis of stem cells. Therefore, we investigated the therapeutic role of adrenaline-stimulated BMSCs on lipopolysaccharide (LPS)-induced lung injury. BMSCs were cultured with adrenergic receptor agonists or antagonists. Suspensions of lung cells or sliced lung tissue from animals with or without LPS-induced injury were co-cultured with BMSCs. LPS-stimulated alveolar macrophages were co-cultured with BMSCs (with adrenaline stimulation or not) in Transwell for 6 hrs. A preliminary animal experiment was conducted to validate the findings in ex vivo study. We found that adrenaline at 10 ?M enhanced proliferation of BMSCs through both ?- and ?-adrenergic receptors. Adrenaline promoted the migration of BMSCs towards LPS-injured lung cells or lung tissue. Adrenaline-stimulated BMSCs decreased the inflammation of LPS-stimulated macrophages, probably through the expression and secretion of several paracrine factors. Adrenaline reduced the extent of injury in LPS-injured rats. Our data indicate that adrenaline-stimulated BMSCs might contribute to the prevention from acute lung injury through the activation of adrenergic receptors, promotion of proliferation and migration towards injured lung, and modulation of inflammation. PMID:24684532

Wu, Xiaodan; Wang, Zhiming; Qian, Mengjia; Wang, Lingyan; Bai, Chunxue; Wang, Xiangdong

2014-01-01

88

Resveratrol Inhibits LPS-Induced MAPKs Activation via Activation of the Phosphatidylinositol 3-Kinase Pathway in Murine RAW 264.7 Macrophage Cells  

PubMed Central

Background Resveratrol is a natural polyphenolic compound that has cardioprotective, anticancer and anti-inflammatory properties. We investigated the capacity of resveratrol to protect RAW 264.7 cells from inflammatory insults and explored mechanisms underlying inhibitory effects of resveratrol on RAW 264.7 cells. Methodology/Principal Findings Murine RAW 264.7 cells were treated with resveratrol (1, 5, and 10 µM) and/or LPS (5 µg/ml). Nitric oxide (NO) and prostaglandin E2 (PGE2) were measured by Griess reagent and ELISA. The mRNA and protein levels of proinflammatory proteins and cytokines were analysed by ELISA, RT-PCR and double immunofluorescence labeling, respectively. Phosphorylation levels of Akt, cyclic AMP-responsive element-binding protein (CREB), mitogen-activated protein kinases (MAPKs) cascades, AMP-activated protein kinase (AMPK) and expression of SIRT1(Silent information regulator T1) were measured by western blot. Wortmannin (1 µM), a specific phosphatidylinositol 3-kinase (PI3-K) inhibitor, was used to determine if PI3-K/Akt signaling pathway might be involved in resveratrol’s action on RAW 264.7 cells. Resveratrol significantly attenuated the LPS-induced expression of nitric oxide (NO), prostaglandin E2 (PGE2), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor-? (TNF-?) and interleukin-1? (IL-1?) in RAW 264.7 cells. Resveratrol increased Akt phosphorylation in a time-dependent manner. Wortmannin, a specific phosphatidylinositol 3-kinase (PI3-K) inhibitor, blocked the effects of resveratrol on LPS-induced RAW 264.7 cells activation. In addition, PI3-K inhibition partially abolished the inhibitory effect of resveratrol on the phosphorylation of cyclic AMP-responsive element-binding protein (CREB) and mitogen-activated protein kinases (MAPKs) cascades. Meanwhile, PI3-K is essential for resveratrol-mediated phosphorylation of AMPK and expression of SIRT1. Conclusion and Implications This investigation demonstrates that PI3-K/Akt activation is an important signaling in resveratrol-mediated activation of AMPK phosphorylation and SIRT1 expression, and inhibition of phosphorylation of CREB and MAPKs activation, proinflammatory mediators and cytokines production in response to LPS in RAW 264.7 cells. PMID:22952890

Liu, Bin; Deng, Yi-Shu; Zhan, Dong; Chen, Yuan-Li; He, Ying; Liu, Jing; Zhang, Zong-Ji; Sun, Jun; Lu, Di

2012-01-01

89

Anti-inflammatory effect of procyanidins from wild grape (Vitis amurensis) seeds in LPS-induced RAW 264.7 cells.  

PubMed

In the present study, the anti-inflammatory effect and underlying mechanisms of wild grape seeds procyanidins (WGP) were examined using lipopolysaccharide- (LPS-) stimulated RAW 264.7 cells. We used nitric oxide (NO) and prostaglandin E2 (PGE2) and reactive oxygen species (ROS) assays to examine inhibitory effect of WGP and further investigated the mechanisms of WGP suppressed LPS-mediated genes and upstream expression by Western blot and confocal microscopy analysis. Our data indicate that WGP significantly reduced NO, PGE2, and ROS production and also inhibited the expression of proinflammatory mediators such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) protein expressions. Consistently, WGP significantly reduced LPS-stimulated expression of proinflammatory cytokines such as tumor necrosis factor ? (TNF-?) and interleukin- (IL-) 1 ? . Moreover, WGP prevented nuclear translocation of nuclear factor- ? B (NF ? B) p65 subunit by reducing inhibitory ? B- ? (I ? B ?) and NF ? B phosphorylation. Furthermore, we found that WGP inhibited LPS-induced phosphorylation of p38 mitogen-activated protein kinase (MAPK). Taken together, our results demonstrated that WGP exerts potent anti-inflammatory activity through the inhibition of iNOS and COX-2 by regulating NF ? B and p38 MAPK pathway. PMID:24260615

Bak, Min-Ji; Truong, Van Long; Kang, Hey-Sook; Jun, Mira; Jeong, Woo-Sik

2013-01-01

90

Anti-Inflammatory Effect of Procyanidins from Wild Grape (Vitis amurensis) Seeds in LPS-Induced RAW 264.7 Cells  

PubMed Central

In the present study, the anti-inflammatory effect and underlying mechanisms of wild grape seeds procyanidins (WGP) were examined using lipopolysaccharide- (LPS-) stimulated RAW 264.7 cells. We used nitric oxide (NO) and prostaglandin E2 (PGE2) and reactive oxygen species (ROS) assays to examine inhibitory effect of WGP and further investigated the mechanisms of WGP suppressed LPS-mediated genes and upstream expression by Western blot and confocal microscopy analysis. Our data indicate that WGP significantly reduced NO, PGE2, and ROS production and also inhibited the expression of proinflammatory mediators such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) protein expressions. Consistently, WGP significantly reduced LPS-stimulated expression of proinflammatory cytokines such as tumor necrosis factor ? (TNF-?) and interleukin- (IL-) 1?. Moreover, WGP prevented nuclear translocation of nuclear factor-?B (NF?B) p65 subunit by reducing inhibitory ?B-? (I?B?) and NF?B phosphorylation. Furthermore, we found that WGP inhibited LPS-induced phosphorylation of p38 mitogen-activated protein kinase (MAPK). Taken together, our results demonstrated that WGP exerts potent anti-inflammatory activity through the inhibition of iNOS and COX-2 by regulating NF?B and p38 MAPK pathway. PMID:24260615

Bak, Min-Ji; Truong, Van Long; Kang, Hey-Sook; Jun, Mira; Jeong, Woo-Sik

2013-01-01

91

Polysaccharides from Smilax glabra inhibit the pro-inflammatory mediators via ERK1/2 and JNK pathways in LPS-induced RAW264.7 cells.  

PubMed

The rhizomes of Smilax glabra have been used as both food and folk medicine in many countries for a long time. However, little research has been reported on polysaccharides of S. glabra. In the present study, two polysaccharide fractions, SGP-1 and SGP-2, were isolated from the rhizomes of S. glabra with the number average molecular weights of 1.72×10(2)kDa and 1.31×10(2)kDa, and the weight average molecular weights of 1.31×10(5)kDa and 1.18×10(5)kDa, respectively, and their mainly monosaccharide compositions were both galactose and rhamnose (2.5:1). Both SGP-1 and SGP-2 significantly suppressed the release of nitric oxide (NO), tumor necrosis factor-? (TNF-?) and interleukin-6 (IL-6) from LPS-induced RAW 264.7 cells, as well as the mRNA expression of inducible nitric oxide synthase (iNOS), TNF-? and IL-6. Additionally, SGP-1 and SGP-2 repressed the extracellular signal-regulated kinase (ERK) and c-Jun NH2-terminal kinase (JNK). These findings strongly suggested polysaccharides were also the anti-inflammatory active ingredient for S. glabra, and the potential of SGP-1 and SGP-2 as the anti-inflammatory agents. PMID:25817687

Chuan-Li, Lu; Wei, Zhu; Min, Wang; Meng-Mei, Hu; Wen-Long, Chen; Xiao-Jie, Xu; Chuan-Jian, Lu

2015-05-20

92

Inhibition of LPS-induced chemokine production in human lung endothelial cells by lipid conjugates anchored to the membrane  

PubMed Central

In acute respiratory distress syndrome (ARDS) induced by endotoxins, a high production of inflammatory mediators by microvascular lung endothelial cells (LMVEC) can be observed. Activation of cells by endotoxins may result in elevated secretion of phospholipase A2 (sPLA2) which is thought to contribute to tissue damage. The present study was undertaken to investigate the role of sPLA2 in chemokine production in human lung microvascular endothelial cells (LMVEC) stimulated with the endotoxins lipopolysaccharide (LPS) and lipoteichoic acid (LTA). In particular, we investigated the effects of sPLA2 inhibitors, specifically, the extracellular PLA2 inhibitors (ExPLIs), composed of N-derivatized phosphatidyl-ethanolamine linked to polymeric carriers, and LY311727, a specific inhibitor of non-pancreatic sPLA2. ExPLIs markedly inhibited LPS and LTA induced production and mRNA expression of the neutrophile attracting chemokines IL-8, Gro-? and ENA-78, as well as of the adhesion molecules ICAM-1 and E-selectin. Concomitantly, ExPLIs inhibited the LPS-induced activation of NF-?B by LPS but not its activation by TNF-? or IL-1. Endotoxin mediated chemokine production in LMVEC seems not to involve PLA2 activity, since LPS stimulation was not associated with activation of intracellular or secreted PLA2. It therefore seems that the inhibitory effect of the ExPLIs was not due to their PLA2 inhibiting capacity. This was supported by the finding that the LPS-induced chemokine production was not affected by the selective sPLA2 inhibitor LY311727. It is proposed that the ExPLIs may be considered a prototype of potent suppressors of specific endotoxin-induced inflammatory responses, with potential implications for the therapy of subsequent severe inflammation. PMID:11934806

Beck, G Ch; Yard, B A; Schulte, J; Oberacker, R; Ackern, K van; Woude, F J van der; Krimsky, M; Kaszkin, M; Yedgar, S

2002-01-01

93

Prostacyclin post-treatment improves LPS-induced acute lung injury and endothelial barrier recovery via Rap1.  

PubMed

Protective effects of prostacyclin (PC) or its stable analog beraprost against agonist-induced lung vascular inflammation have been associated with elevation of intracellular cAMP and Rac GTPase signaling which inhibited the RhoA GTPase-dependent pathway of endothelial barrier dysfunction. This study investigated a distinct mechanism of PC-stimulated lung vascular endothelial (EC) barrier recovery and resolution of LPS-induced inflammation mediated by small GTPase Rap1. Efficient barrier recovery was observed in LPS-challenged pulmonary EC after prostacyclin administration even after 15h of initial inflammatory insult and was accompanied by the significant attenuation of p38 MAP kinase and NF?B signaling and decreased production of IL-8 and soluble ICAM1. These effects were reproduced in cells post-treated with 8CPT, a small molecule activator of Rap1-specific nucleotide exchange factor Epac. By contrast, pharmacologic Epac inhibitor, Rap1 knockdown, or knockdown of cell junction-associated Rap1 effector afadin attenuated EC recovery caused by PC or 8CPT post-treatment. The key role of Rap1 in lung barrier restoration was further confirmed in the murine model of LPS-induced acute lung injury. Lung injury was monitored by measurements of bronchoalveolar lavage protein content, cell count, and Evans blue extravasation and live imaging of vascular leak over 6days using a fluorescent tracer. The data showed significant acceleration of lung recovery by PC and 8CPT post-treatment, which was abrogated in Rap1a(-/-) mice. These results suggest that post-treatment with PC triggers the Epac/Rap1/afadin-dependent mechanism of endothelial barrier restoration and downregulation of p38MAPK and NF?B inflammatory cascades, altogether leading to accelerated lung recovery. PMID:25545047

Birukova, Anna A; Meng, Fanyong; Tian, Yufeng; Meliton, Angelo; Sarich, Nicolene; Quilliam, Lawrence A; Birukov, Konstantin G

2015-05-01

94

Identification and expression of a putative LPS-induced TNF-? factor from Asiatic hard clam Meretrix meretrix.  

PubMed

LPS-induced TNF-? (LITAF) is a novel transcriptional factor that mediates the expression of inflammatory cytokines in LPS-induced processes. In the present study, the full-length cDNA encoding LITAF (designated as Mm-LITAF) was identified from Asiatic hard clam, Meretrix meretrix, by expressed sequence tag and rapid amplification of cDNA ends (RACE) approaches. The full-length cDNA of Mm-LITAF was 1653 bp, consisting of a 5' untranslated region (UTR) of 91 bp, a 3'UTR of 1166 bp with one cytokine RNA instability motif (ATTTA) and one polyadenylation signal (AATAAA), and an open reading frame (ORF) of 396 bp encoding a polypeptide of 131 amino acids with a theoretical isoelectric point of 7.49, and predicted molecular weight of 14.47 kDa. The deduced amino acid of Mm-LITAF shared 29-63% similarity with the LITAFs from other species, indicating that Mm-LITAF should be a member of the LITAF family. Two highly conserved CXXC motifs forming a compact Zn(2+)-binding structure were also identified in Mm-LITAF. A quantitative reverse transcriptase real-time PCR (qRT-PCR) assay was developed to assess the expression of Mm-LITAF mRNA in different tissues, and the temporal expression of Mm-LITAF in clams challenged with Vibrio anguillarum. The mRNA transcript of Mm-LITAF could be detected in all the examined tissues with the highest expression level in the gill. Mm-LITAF expression was up-regulated significantly at 16 h in the gill and at 8 h in haemocytes after bacterial challenge, respectively. These results suggest that the Mm-LITAF is a constitutive and inducible acute-phase protein that perhaps involved in the innate immune response of hard clam. PMID:21567197

Li, Hong-Jun; Yang, Qing; Gao, Xiang-Gang; Su, Hao; Wang, Juan; He, Chong-Bo

2012-02-01

95

Selective inhibition of sphingosine kinase-1 protects adipose tissue against LPS-induced inflammatory response in Zucker diabetic fatty rats.  

PubMed

Obesity is associated with a state of chronic inflammation. The chemokine (C-C motif) ligand 5 (CCL5) has been proposed to modulate the inflammatory response in adipose tissue (AT). However, the mechanisms underlying CCL5 upregulation in AT remain undefined. The objective of the present study was to evaluate whether the enzyme sphingosine kinase-1 (SK1) would modulate the expression of CCL5 and other inflammatory biomarkers in primary adipocytes and its potential role in lipopolysaccharide (LPS)-induced AT inflammation in a rat model of diabetes. To address this, LPS-stimulated primary adipocytes and 3T3-L1 cells were treated with a SK inhibitor, and the expression of Ccl5 and other CC chemokines were studied. Moreover, the effect of SK1 knockdown on cytokine production was analyzed in 3T3-L1 cells by transfection of SK1-specific small-interfering RNA (siRNA). The anti-inflammatory effects of SK inhibitor in AT were also investigated in vivo using the Zucker lean normoglycemic control (ZLC) rats. LPS treatment stimulated Ccl5, IL-6, pentraxin 3 (Ptx3), and Tnf? mRNA expression in primary adipocytes and 3T3-L1 cells, whereas pharmacologically and siRNA-mediated SK1 inhibition strongly reduced mRNA levels of proinflammatory cytokines in these cells. Similarly, administration of SK inhibitor to ZLC rats prevented the LPS-induced inflammatory response in AT. Our data demonstrate a role for SK1 in endotoxin-induced cytokine expression in adipocytes and suggest that inhibition of SK1 may be a potential therapeutic tool in the prevention and treatment of chronic and common metabolic disorders, including obesity, insulin-resistance, and type 2 diabetes. PMID:25053402

Tous, Monica; Ferrer-Lorente, Raquel; Badimon, Lina

2014-09-01

96

MyD88–BLT2-dependent cascade contributes to LPS-induced interleukin-6 production in mouse macrophage  

PubMed Central

Endotoxic responses to bacterial lipopolysaccharide (LPS) are triggered by Toll-like receptor 4 (TLR4) and involve the production of inflammatory mediators, including interleukin-6 (IL-6), by macrophages. The detailed mechanism of IL-6 production by macrophages in response to LPS has remained unclear, however. We now show that LPS induces IL-6 synthesis in mouse peritoneal macrophages via the leukotriene B4 receptor BLT2. Our results suggest that TLR4–MyD88 signaling functions upstream of BLT2 and that the generation of reactive oxygen species (ROS) by NADPH oxidase 1 (Nox1) and consequent activation of the transcription factor nuclear factor (NF)-?B function downstream of BLT2 in this response. These results suggest that a TLR4–MyD88–BLT2–Nox1–ROS–NF-?B pathway contributes to the synthesis of IL-6 in LPS-stimulated mouse macrophages.

Lee, A-Jin; Cho, Kyung-Jin; Kim, Jae-Hong

2015-01-01

97

MyD88-BLT2-dependent cascade contributes to LPS-induced interleukin-6 production in mouse macrophage.  

PubMed

Endotoxic responses to bacterial lipopolysaccharide (LPS) are triggered by Toll-like receptor 4 (TLR4) and involve the production of inflammatory mediators, including interleukin-6 (IL-6), by macrophages. The detailed mechanism of IL-6 production by macrophages in response to LPS has remained unclear, however. We now show that LPS induces IL-6 synthesis in mouse peritoneal macrophages via the leukotriene B4 receptor BLT2. Our results suggest that TLR4-MyD88 signaling functions upstream of BLT2 and that the generation of reactive oxygen species (ROS) by NADPH oxidase 1 (Nox1) and consequent activation of the transcription factor nuclear factor (NF)-?B function downstream of BLT2 in this response. These results suggest that a TLR4-MyD88-BLT2-Nox1-ROS-NF-?B pathway contributes to the synthesis of IL-6 in LPS-stimulated mouse macrophages. PMID:25838003

Lee, A-Jin; Cho, Kyung-Jin; Kim, Jae-Hong

2015-01-01

98

The regulation of cytochrome P450 2E1 during LPS-induced inflammation in the rat  

SciTech Connect

It is well known that inflammatory and infectious conditions differentially regulate cytochrome P450 (P450)-mediated drug metabolism in the liver. We have previously outlined a potential pathway for the downregulation in hepatic cytochrome P450 following LPS-mediated inflammation in the CNS (Abdulla, D., Goralski, K.B., Garcia Del Busto Cano, E., Renton, K.W., 2005. The signal transduction pathways involved in hepatic cytochrome P450 regulation in the rat during an LPS-induced model of CNS inflammation. Drug Metab. Dispos). The purpose of this study was to outline the effects of LPS-induced peripheral and central nervous system inflammation on hepatic cytochrome P450 2E1 (CYP2E1) in vivo, an enzyme that plays an important role in various physiological and pathological states. We report an increase in hepatic mRNA expression of CYP2E1 that occurred as early as 2-3 h following either the intraperitoneal (i.p.) injection of 5 mg/kg LPS or i.c.v. administration of 25 {mu}g of LPS. This increase in CYP2E1 mRNA expression was sustained for 24 h. In sharp contrast to the increase in hepatic CYP2E1 mRNA, we observed a significant reduction in the catalytic activity of this enzyme 24 h following either the i.c.v. or i.p. administration of LPS. Cycloheximide or actinomycin-D did not change the LPS-mediated downregulation in hepatic CYP2E1 catalytic activity. Our results support the idea that LPS acts at two different levels to regulate hepatic CYP2E1: a transcriptional level to increase CYP2E1 mRNA expression and a post-transcriptional level to regulate CYP2E1 protein and activity.

Abdulla, Dalya [Department of Pharmacology, Sir Charles Tupper Medical Bldg., Dalhousie University, Halifax, Nova Scotia, B3H 4H7 (Canada); Goralski, Kerry B. [Department of Pharmacology, Sir Charles Tupper Medical Bldg., Dalhousie University, Halifax, Nova Scotia, B3H 4H7 (Canada); College of Pharmacy, Burbidge Building, Dalhousie University, Halifax, Nova Scotia, B3H 3J5 (Canada); Renton, Kenneth W. [Department of Pharmacology, Sir Charles Tupper Medical Bldg., Dalhousie University, Halifax, Nova Scotia, B3H 4H7 (Canada)]. E-mail: Ken.Renton@dal.ca

2006-10-01

99

LPS-Induced Lung Inflammation in Marmoset Monkeys – An Acute Model for Anti-Inflammatory Drug Testing  

PubMed Central

Increasing incidence and substantial morbidity and mortality of respiratory diseases requires the development of new human-specific anti-inflammatory and disease-modifying therapeutics. Therefore, new predictive animal models that closely reflect human lung pathology are needed. In the current study, a tiered acute lipopolysaccharide (LPS)-induced inflammation model was established in marmoset monkeys (Callithrix jacchus) to reflect crucial features of inflammatory lung diseases. Firstly, in an ex vivo approach marmoset and, for the purposes of comparison, human precision-cut lung slices (PCLS) were stimulated with LPS in the presence or absence of the phosphodiesterase-4 (PDE4) inhibitor roflumilast. Pro-inflammatory cytokines including tumor necrosis factor-alpha (TNF-?) and macrophage inflammatory protein-1 beta (MIP-1?) were measured. The corticosteroid dexamethasone was used as treatment control. Secondly, in an in vivo approach marmosets were pre-treated with roflumilast or dexamethasone and unilaterally challenged with LPS. Ipsilateral bronchoalveolar lavage (BAL) was conducted 18 hours after LPS challenge. BAL fluid was processed and analyzed for neutrophils, TNF-?, and MIP-1?. TNF-? release in marmoset PCLS correlated significantly with human PCLS. Roflumilast treatment significantly reduced TNF-? secretion ex vivo in both species, with comparable half maximal inhibitory concentration (IC50). LPS instillation into marmoset lungs caused a profound inflammation as shown by neutrophilic influx and increased TNF-? and MIP-1? levels in BAL fluid. This inflammatory response was significantly suppressed by roflumilast and dexamethasone. The close similarity of marmoset and human lungs regarding LPS-induced inflammation and the significant anti-inflammatory effect of approved pharmaceuticals assess the suitability of marmoset monkeys to serve as a promising model for studying anti-inflammatory drugs. PMID:22952743

Seehase, Sophie; Lauenstein, Hans-Dieter; Schlumbohm, Christina; Switalla, Simone; Neuhaus, Vanessa; Förster, Christine; Fieguth, Hans-Gerd; Pfennig, Olaf; Fuchs, Eberhard; Kaup, Franz-Josef; Bleyer, Martina; Hohlfeld, Jens M.; Braun, Armin

2012-01-01

100

Cytokines and LPS induce iNOS but not Enzyme Activity in Adult Rat Cardiomyocytes  

PubMed Central

There is evidence that nitric oxide (NO) formation in adult cardiomyocytes stimulated with lipopolysaccharide (LPS) does not commensurate with iNOS levels. Tetrahydrobiopterin (BH4) is a key factor in the stabilization and NO production by iNOS homodimer. Thus we hypothesized that BH4 is a limiting factor for NO production in adult cardiomyocytes in response to LPS and cytokines (TNF-?, IL-1, IFN-? alone or mixed). It was verified that LPS and cytokines induced iNOS expression which did not translate into increased nitrite or 14C-citrulline production. This response coincided with defective BH4 synthesis and low GTP cyclohydrolase activity. Furthermore, supplementation with BH4 and ascorbate failed to increase iNOS activity. This effect was related to preferential accumulation of BH2 rather than BH4 in these cells. Uncoupled iNOS activity in stimulated cells was examined using mitochondrial aconitase activity as an endogenous marker of superoxide anion radical (O2•?) formation, and found not to be significantly inhibited. 2-Hydroxyethidium also showed not to be significantly increased. We conclude that adult cardiomyocytes are an unlikely source of NO and O2•? in inflammatory conditions. This finding adds a new and unexpected layer of complexity to our understanding of the responses of the adult heart to inflammation. PMID:18634867

Vásquez-Vivar, Jeannette; Whitsett, Jennifer; Ionova, Irina; Konorev, Eugene; Zielonka, Jacek; Kalyanaraman, Balaraman; Shi, Yang; Pieper, Galen M.

2008-01-01

101

Gonadotropin-releasing hormone agonist selectively augments thymopoiesis and prevents cell apoptosis in LPS induced thymic atrophy model independent of gonadal steroids.  

PubMed

Lipopolysaccharide (LPS) causes acute thymic atrophy, a phenomenon that has been linked to immune dysfunction and poor survival during sepsis. The systemic response to LPS involves a rise in glucocorticoids and proinflammatory cytokines which contribute greatly to thymic involution and apoptosis. Gonadotropin-releasing hormone (GnRH) analog exerts thymopoietic regulatory effects and possesses immunostimulant properties. We determined whether leuprolide, a GnRH analog can be useful in LPS induced thymic involution and apoptosis. Mice injected with 100 ?g of LPS intraperitoneally led to involution of thymus, to decrease of CD4(+)8(+) thymocyte subset, and to fragmentation of thymic DNA. Leuprolide (100 ?g/mouse, s.c.) pretreatment significantly attenuated LPS induced thymic atrophy, and also reduced LPS induced systemic rise in corticosterone levels. The observed effect of leuprolide remained unaffected in castrated and ovariectomized mice. Collectively, leuprolide has protective action independent of gonadal steroids, which was mediated by blunting of the systemic corticosteroid response in LPS induced thymic atrophy model. PMID:25111851

Ullewar, Meenal P; Umathe, Sudhir N

2014-11-01

102

Santamarin, a sesquiterpene lactone isolated from Saussurea lappa, represses LPS-induced inflammatory responses via expression of heme oxygenase-1 in murine macrophage cells.  

PubMed

Saussurea lappa C.B. Clarke (Compositae) is indigenous to India and Pakistan. The dried root of S. lappa has been traditionally used for alleviating pain in abdominal distention and tenesmus, indigestion with anorexia, dysentery, nausea, and vomiting. Santamarin is a sesquiterpene lactone isolated from S. lappa. In the present study, santamarin inhibited inducible nitric oxide synthase (iNOS) protein, reduced iNOS-derived nitric oxide (NO), suppressed COX-2 protein and reduced COX-derived PGE(2) production in lipopolysaccharide (LPS)-stimulated RAW264.7 cells and murine peritoneal macrophages. Similarly, santamarin reduced tumor necrosis factor-? (TNF-?) and interleukin-1? (IL-1?) production. In addition, santamarin suppressed the phosphorylation and degradation of I?B-? as well as the nuclear translocation of p65 in response to LPS in RAW264.7 cells. Furthermore, santamarin induced heme oxygenase (HO)-1 expression mRNA and protein level that plays a cytoprotective role against inflammation. The induction of HO-1 is primarily regulated at the transcriptional level, and its induction by various agents is mediated by the nuclear transcription factor E2-related factor 2 (Nrf2), master regulator of antioxidant responses. Unbound Nrf2 translocates into the nucleus and binds to the antioxidant response element (ARE) in the upstream promoter region of many antioxidative genes, where it initiates their transcription. The effects of santamarin on LPS-induced NO, PGE(2), TNF-?, and IL-1? production were partially reversed by the HO-1 inhibitor, tin protoporphyrin (SnPP). Therefore, our data suggest that the anti-inflammatory effect of santamarin in macrophages may be exerted through a novel mechanism that involves HO-1 expression. PMID:22564506

Choi, Hyun-Gyu; Lee, Dong-Sung; Li, Bin; Choi, Yeon Ho; Lee, Seung-Ho; Kim, Youn-Chul

2012-07-01

103

SIGNR1-mediated phagocytosis, but not SIGNR1-mediated endocytosis or cell adhesion, suppresses LPS-induced secretion of IL-6 from murine macrophages.  

PubMed

C-type lectin receptors (CLRs) serve as phagocytosis receptors for pathogens and also function as adhesion molecules and in the recognition and endocytosis of glycosylated self-antigens. In the present study, we demonstrated that phagocytosis mediated by a mouse mannose-binding CLR, SIGNR1 significantly suppressed the LPS-induced secretion of the specific pro-inflammatory cytokines from the resident peritoneal macrophages and the mouse macrophage-like cells that express SIGNR1 (RAW-SIGNR1). LPS-induced secretion of IL-6 from peritoneal macrophages suppressed in response to uptake of oligomannose-coated liposomes (OMLs), and the suppression was partly inhibited by treatment with an anti-SIGNR1 antibody. LPS-induced secretion of IL-6 from RAW-SIGNR1 cells was also clearly inhibited by treatment of the cells with OMLs >0.4?m in diameter, but treatment with OMLs <0.4?m in diameter did not affect the IL-6 secretion. In contrast, LPS-induced TNF-? secretion from the cells was not affected on treatment of the cells with OMLs. Suppression of the IL-6 secretion was not observed following treatment with oligomannose-containing soluble polymers or when cells were bound to an oligomannose-coated solid phase. Phagocytosis of oligomannose-coated liposomes did not interfere with the transcription of IL-6 mRNA, but did affect IL-6 mRNA stability, leading to suppression of IL-6 secretion. Interestingly, treatment of the cells with Ly290042, a PI3 kinase inhibitor, partly blocked the suppression of LPS-induced secretion of IL-6 by OML. Thus, we conclude that SIGNR1-mediated phagocytosis but not SIGNR1-mediated endocytosis and cell adhesion, suppresses the TLR4-mediated production of specific proinflammatory cytokines via PI3 kinase signaling. PMID:25226443

Kawauchi, Yoko; Takagi, Hideaki; Hanafusa, Kei; Kono, Mirei; Yamatani, Minami; Kojima, Naoya

2015-01-01

104

High-mobility group box-1 was released actively and involved in LPS induced depressive-like behavior.  

PubMed

Depression disorder is a common mental illness, of which the pathogenesis is not well understood. Studies suggest that immunity imbalance and up-regulation of pro-inflammatory cytokines may be associated with the pathogenesis of depression. High-mobility group box 1 protein (HMGB1) has gained much attention as an important player in innate immune responses and an modulating factor in several inflammatory diseases. Here we sought to explore the role of HMGB1 in the development of depression. Depression model was established with low dose of lipopolysaccharide (LPS) administration. Depressive behavior was reflected with increased immobility time in tail suspension test. Accompanying with depressive-like behavior, translocation of HMGB1 from nuclei to cytoplasm was observed by immunofluorescence assays. Meanwhile, no significant necrosis was observed evaluated by hematoxylin-eosin staining. These data indicated that HMGB1 was released actively in the central nervous system. In addition, treating the mice with human recombinant HMGB1 (rHMGB1) could induce the development of depressive-like behavior. Blockage of HMGB1 with GZA abrogated the depressive-like behavior induced by LPS or rHMGB1. These results implicated that HMGB1 was involved in LPS-induced depressive-like behavior. PMID:25795092

Wu, Teng-Yun; Liu, Lei; Zhang, Wei; Zhang, Yi; Liu, Yun-Zi; Shen, Xiao-Liang; Gong, Hong; Yang, Yuan-Yuan; Bi, Xiao-Ying; Jiang, Chun-Lei; Wang, Yun-Xia

2015-05-01

105

Nicotine exaggerates LPS-induced airway hyperreactivity via JNK-mediated up-regulation of Toll-like receptor 4.  

PubMed

Tobacco smokers often display increased airway hyperreactivity (AHR) when faced with bacterial infections. The present study uses a murine organ-culture model to dissect the mechanisms involved in this exaggerated smooth muscle response. Nicotine simulates the effects of smoking, and LPS represents bacterial infection. Contractile responses of isolated murine tracheal segments were analyzed in myographs after organ culture with increasing concentrations of LPS and/or nicotine for 4 days with or without specific MAPK inhibitors. Nicotine's effect on the expression of cell surface Toll-like receptors (TLRs), MCP-1, COX-2, and TNF-? were examined by real-time PCR. Increased protein expression was verified by immunohistochemistry. LPS concentration-dependently increased contractile responses to bradykinin and des-Arg(9)-bradykinin. A combination of nicotine and low-dose LPS caused powerful synergistic contractions along with increased kinin receptor expression. Specific kinin B1 and B2 receptor inhibitors blocked this reaction. Nicotine increased mRNA and protein expression of TLR4 and -6 in the epithelium and smooth muscle layer, with MCP-1 and COX-2 mRNA increasing in parallel. Specific inhibition of JNK attenuated nicotine's effects. In conclusion, long-term exposure to nicotine up-regulated the expression of TLR4 and -6 via a JNK-related pathway, causing an exaggeration of the LPS-induced local airway inflammation and increased AHR. This might offer a mechanistic explanation to the increased AHR seen in tobacco smokers confronted with bacterial infections. PMID:24669857

Xu, Yuan; Zhang, Yaping; Cardell, Lars-Olaf

2014-09-01

106

Ultrapure LPS induces inflammatory and antibacterial responses attenuated in vitro by exogenous sera in Atlantic cod and Atlantic salmon.  

PubMed

Phagocyte recognition of lipopolysaccharide (LPS) is an early key event for triggering the host innate immune response necessary for clearance of invading bacteria. The ability of fishes to recognise LPS has been questioned as contradictory results have been presented. We show here that monocyte/macrophage cultures from Atlantic cod (Gadus morhua) and Atlantic salmon (Salmo salar) respond with an increased expression of inflammatory and antibacterial genes to both crude and ultrapure Escherichia coli LPS. Crude LPS produces higher induction than the ultrapure LPS type in both species in vitro as well as in vivo in cod injected with LPS. Crude LPS gave, in contrast to ultrapure LPS, an additional weak up-regulation of antiviral genes in salmon macrophages, most likely because of contaminants in the LPS preparation. Increased levels of chicken (c)-type lysozyme transcripts and enzyme activity were measured in salmon macrophages following ultrapure LPS stimulation demonstrating not only increased transcription but also translation. Simultaneous use and even pre-treatment with bovine sera suppressed the LPS-induced expression thereby reflecting the presence of transcription inhibitory components in sera. Together, these findings show that both cod and salmon recognise LPS per se and that the observed induction is highly dependent on the absence of sera. PMID:25655332

Seppola, Marit; Mikkelsen, Helene; Johansen, Audny; Steiro, Kari; Myrnes, Bjørnar; Nilsen, Inge W

2015-05-01

107

Microbial transformation of acetyl-11-keto-?-boswellic acid and their inhibitory activity on LPS-induced NO production.  

PubMed

The capabilities of 20 strains of fungi to transform acetyl-11-keto-?-boswellic (AKBA) were screened. And biotransformation of AKBA by Cunninghamella blakesleana AS 3.970 afforded five metabolites (1-5), while two metabolites (6, 7) were isolated from biotransformation of Cunninghamella elegans AS 3.1207. The chemical structures of these metabolites were identified by spectral methods including 2D NMR and their structures were elucidated as 7?-hydroxy-3-acety-11-keto-?-boswellic acid (1), 21?-dihydroxy-3-acety-11-keto-?-boswellic acid (2), 7?,22?-dihydroxy-3-acety-11-keto-?-boswellic acid (3), 7?,16?-dihydroxy-3-acety-11-keto-?-boswellic acid (4), 7?,15?-dihydroxy-3-acety-11-keto-?-boswellic acid (5); 7?,15?,21?-trihydroxy-3-acety-11-keto-?-boswellic acid (6) and 15?,21?-dihydroxy-3-acety-11-keto-?-boswellic acid (7). All these products are previously unknown. Their primary structure-activity relationships (SAR) of inhibition activity on LPS-induced NO production in RAW 264.7 macrophage cells were evaluated. PMID:23391590

Sun, Yan; Liu, Dan; Xi, Ronggang; Wang, Xiaobo; Wang, Yan; Hou, Jie; Zhang, Baojing; Wang, Changyuan; Liu, Kexin; Ma, Xiaochi

2013-03-01

108

FPR2/ALX activation reverses LPS-induced vascular hyporeactivity in aorta and increases survival in a pneumosepsis model.  

PubMed

The formylpeptide receptor 2 (FPR2/ALX) is a very promiscuous receptor, utilized by lipid and protein ligands that trigger pro- or anti-inflammatory responses. FPR2/ALX expression is increased in lung tissues of septic animals and its activation has a beneficial therapeutic effect by controlling exacerbated inflammation. Although FPR2/ALX expression was observed in vascular smooth muscle cells, its role in vascular reactivity in inflammatory conditions has not been studied. In this study, we report that LPS increases FPR2/ALX expression in vascular smooth muscle cells (A7r5 cells) and aorta tissue, and that the selective agonist WKYMVm reverses LPS-induced vascular hyporeactivity in mouse aorta rings. Mice bearing pneumosepsis by Klebsiella pneumoniae and treated with WKYMVm recovered the reactivity to vasoconstrictors and the survival improved by 40%. As for the mechanisms involved, FPR2/ALX activation decreases NO production in LPS-stimulated cells and aorta, but it does not seem involve the regulation of NOS-2 expression. The molecular mechanism by which the peptide inhibits NO production still needs to be elucidated, but our data suggests an important role for NO in the WKYMVm beneficial effect observed in LPS injury and sepsis. In conclusion, our data suggest, for the first time, that a receptor, primarily described as a mediator of immune responses, may have an important role in the vascular dysfunctions observed in sepsis and may be a possible target for new therapeutic interventions. PMID:25478948

Horewicz, Verônica Vargas; Crestani, Sandra; de Sordi, Regina; Rezende, Edir; Assreuy, Jamil

2015-01-01

109

Wogonin Inhibits LPS-Induced Inflammatory Responses in Rat Dorsal Root Ganglion Neurons Via Inhibiting TLR4-MyD88-TAK1-Mediated NF-?B and MAPK Signaling Pathway.  

PubMed

Recent studies showed that the activation of toll-like receptor 4 (TLR4) on dorsal root ganglion (DRG) neurons might underlie neuropathic and inflammatory pain states. This study was undertaken to investigate the effects of wogonin, a flavonoid with potent anti-inflammatory properties on the inflammatory reaction and TLR4 dependent pathways in lipopolysaccharide (LPS)-treated DRG neurons. Our results showed that wogonin not only inhibited the expression and interaction of TLR4, MyD88, and TAK1, but also reduced the activation of nuclear factor kappa B and mitogen-activated protein kinases pathway in LPS-treated DRG neurons. Moreover, wogonin significantly suppressed the release of pro-inflammatory mediators in LPS-induced DRG neurons, including cyclooxygenase-2, inducible nitric oxide synthases, interleukin (IL)-1?, IL-6, and tumor necrosis factor-alpha. Our results suggested that pre-treatment with wogonin could attenuate the TLR4-mediated inflammatory response in LPS-induced DRG neurons, thus might be beneficial for the treatment of neuropathic and inflammatory pain. PMID:25504431

Chen, Shibiao; Xiong, Jiangqin; Zhan, Yanping; Liu, Weicheng; Wang, Xiuhong

2015-05-01

110

GATA-2 Transduces LPS-Induced il-1? Gene Expression in Macrophages via a Toll-Like Receptor 4/MD88/MAPK-Dependent Mechanism  

PubMed Central

Lipopolysaccharide (LPS) is a critical factor for inducing acute lung injury. GATA-2, a transcription factor, contributes to the control of cell activity and function. Exposure of RAW 264.7 cells to LPS induced interleukin (IL)-1? mRNA and protein expression and GATA-2 translocation from the cytoplasm to nuclei in concentration- and time-dependent manners. A bioinformatic search revealed that GATA-2-specific binding elements exist in the 5’-promoter region of the il-1? gene. LPS could enhance the transactivation activity of GATA-2 in macrophages. Knocking-down translation of GATA-2 mRNA using RNA interference significantly alleviated LPS-induced IL-1? mRNA and protein expression. As to the mechanism, transfection of toll-like receptor (TLR) 4 small interfering (si)RNA into macrophages concurrently decreased LPS-caused increases in nuclear GATA-2 levels. Sequentially, treatment with myeloid differentiation factor 88 (MyD88) siRNA decreased LPS-induced phosphorylation of mitogen-activated protein kinases (MAPKs) kinase 1/2 and subsequent translocation of GATA-2. Reducing MAPK activities using specific inhibitors simultaneously decreased GATA-2 activation. Furthermore, exposure of primary macrophages to LPS significantly increased the transactivation activities of GATA-2 and IL-1? mRNA and protein expression. Transfection of GATA-2 siRNA inhibited LPS-induced IL-1? mRNA expression. Results of this study show that LPS induction of il-1? gene expression in macrophages is mediated by GATA-2 via activation of TLR4, MyD88, and MAPKs. PMID:23940812

Wu, Tsu-Tuan; Tai, Yu-Ting; Cherng, Yih-Giun; Chen, Tyng-Guey; Lin, Chien-Ju; Chen, Ta-Liang; Chang, Huai-Chia; Chen, Ruei-Ming

2013-01-01

111

POLYCHLORINATED BIPHENYL MIXTURES (AROCLORS) INHIBIT LPS-INDUCED MURINE SPLENOCYTE PROLIFERATION IN VITRO. (R826687)  

EPA Science Inventory

Abstract The immune system is believed to be a sensitive indicator for adverse polychlorinated biphenyl (PCB)-induced health effects. Four commercial PCB mixtures (Aroclors) or six individual PCB congeners were evaluated for their effect on splenocyte viability and lip...

112

The Probiotic Mixture VSL#3 Dampens LPS-Induced Chemokine Expression in Human Dendritic Cells by Inhibition of STAT-1 Phosphorylation  

PubMed Central

VSL#3, a mixture of 8 different probiotic bacteria, has successfully been used in the clinic to treat Ulcerative Colitis. We previously identified the modulation of chemokines as a major mechanism in the protective effect of the VSL#3 in a mouse model of colitis. This was supported by in vitro studies that implicated a role for VSL#3 in the suppression of LPS-induced chemokine production by mouse bone marrow-derived dendritic cells (DC). Herein, we validated these findings employing human monocyte-derived DC. Stimulation of human DC with LPS, VSL#3, or a combination of both resulted in their maturation, evident from enhanced expression of activation markers on the cell-surface, as well as the induction of various chemokines and cytokines. Interestingly, a set of LPS-induced chemokines was identified that were suppressed by VSL#3. These included CXCL9, CXCL10, CCL2, CCL7, and CCL8. In silico approaches identified STAT-1 as a dominant regulator of these chemokines, and this was confirmed by demonstrating that LPS-induced phosphorylation of this transcription factor was inhibited by VSL#3. This indicates that VSL#3 may contribute to the control of inflammation by selective suppression of STAT-1 induced chemokines. PMID:25546330

Mariman, Rob; Tielen, Frans; Koning, Frits; Nagelkerken, Lex

2014-01-01

113

Therapeutic effect of intravenous infusion of perfluorocarbon emulsion on LPS-induced acute lung injury in rats.  

PubMed

Acute lung injury (ALI) and its more severe form, acute respiratory distress syndrome (ARDS) are the leading causes of death in critical care. Despite extensive efforts in research and clinical medicine, mortality remains high in these diseases. Perfluorocarbon (PFC), a chemical compound known as liquid ventilation medium, is capable of dissolving large amounts of physiologically important gases (mainly oxygen and carbon dioxide). In this study we aimed to investigate the effect of intravenous infusion of PFC emulsion on lipopolysaccharide (LPS) induced ALI in rats and elucidate its mechanism of action. Forty two Wistar rats were randomly divided into three groups: 6 rats were treated with saline solution by intratracheal instillation (control group), 18 rats were treated with LPS by intratracheal instillation (LPS group) and the other 18 rats received PFC through femoral vein prior to LPS instillation (LPS+PFC group). The rats in the control group were sacrificed 6 hours later after saline instillation. At 2, 4 and 6 hours of exposure to LPS, 6 rats in the LPS group and 6 rats in LPS+PFC group were sacrificed at each time point. By analyzing pulmonary pathology, partial pressure of oxygen in the blood (PaO2) and lung wet-dry weight ratio (W/D) of each rat, we found that intravenous infusion of PFC significantly alleviated acute lung injury induced by LPS. Moreover, we showed that the expression of pulmonary myeloperoxidase (MPO), intercellular adhesion molecule-1 (ICAM-1) of endothelial cells and CD11b of polymorphonuclear neutrophils (PMN) induced by LPS were significantly decreased by PFC treatment in vivo. Our results indicate that intravenous infusion of PFC inhibits the infiltration of PMNs into lung tissue, which has been shown as the core pathogenesis of ALI/ARDS. Thus, our study provides a theoretical foundation for using intravenous infusion of PFC to prevent and treat ALI/ARDS in clinical practice. PMID:24489970

Hou, Shike; Ding, Hui; Lv, Qi; Yin, Xiaofeng; Song, Jianqi; Landén, Ning Xu; Fan, Haojun

2014-01-01

114

Pretreatment of lipopolysaccharide (LPS) ameliorates D-GalN/LPS induced acute liver failure through TLR4 signaling pathway  

PubMed Central

Endotoxin tolerance (ET) is an important phenomenon, which affects inflammation and phagocytosis. Pretreatment with low dose of lipopolysaccharide (LPS) can protect liver injury from various hepatotoxicants such as acetaminophen and pseudomonas aeruginosa exotoxin A. The current study aimed to investigate the protecting mechanisms of endotoxin tolerance in acute liver failure induced by D-galactosamine (D-GalN)/LPS and possible role of toll-like receptors 4 (TLR4) signaling pathway in this phenomenon. Acute liver failure was induced by Injection of D-GalN/LPS. To mimic endotoxin tolerance, male Sprague-Dawley rats were treated with low dose of LPS (0.1 mg/kg once a day intraperitoneally for consecutive five days) before subsequent injection of D-GalN/LPS. Rat survival was determined by survival rate. Liver injury was confirmed by serum biochemical and liver histopathological examination. Inflammatory cytokines were determined by ELISA and nuclear factor-kappa B (NF-?B) (P65), toll-like receptors 4 (TLR4) and Interleukin-1 receptor-associated kinase-1 (IRAK-1) were measured by reverse transcriptase polymerase chain reaction and western blot respectively. Pretreatment of LPS significantly improved rat survival. Moreover, rats pretreated with LPS exhibited lower serum enzyme (ALT, AST and TBiL) level, lower production of inflammatory cytokines and more minor liver histopathological damage than rats without pretreatment of LPS. LPS pretreatment suppressed production of TLR4 and IRAK-1. LPS pretreatment also inhibited activation of hepatic NF-?B. These results indicated that endotoxin tolerance contributed to liver protection against D-GalN/LPS induced acute liver failure through down-regulation of TLR4 and NF-?B pathway. PMID:25400741

Zhang, Sainan; Yang, Naibin; Ni, Shunlan; Li, Wenyuan; Xu, Lanman; Dong, Peihong; Lu, Mingqin

2014-01-01

115

The influence of temperament on lipopolysaccharide (LPS) induced secretion of epinephrine and cortisol in bulls.  

Technology Transfer Automated Retrieval System (TEKTRAN)

The host's complex reaction to a pathogenic stressor involves interaction of the neural, endocrine, and immune systems. For example, exposure to bacteria stimulates secretion of the stress-related hormones, cortisol (CS) and epinephrine (Epi; 1). Innate and induced secretion of CS and Epi are influe...

116

Toona sinensis Inhibits LPS-Induced Inflammation and Migration in Vascular Smooth Muscle Cells via Suppression of Reactive Oxygen Species and NF-?B Signaling Pathway  

PubMed Central

Toona sinensis is one of the most popular vegetarian cuisines in Taiwan and it has been shown to possess antioxidant, antiangiogenic, and anticancer properties. In this study, we investigated the antiatherosclerotic potential of aqueous leaf extracts from Toona sinensis (TS; 25–100??g/mL) and its major bioactive compound, gallic acid (GA; 5??g/mL), in LPS-treated rat aortic smooth muscle (A7r5) cells. We found that pretreatment with noncytotoxic concentrations of TS and GA significantly inhibited inflammatory NO and PGE2 production by downregulating their precursors, iNOS and COX-2, respectively, in LPS-treated A7r5 cells. Furthermore, TS and GA inhibited LPS-induced intracellular ROS and their corresponding mediator, p47phox. Notably, TS and GA pretreatment significantly inhibited LPS-induced migration in transwell assays. Gelatin zymography and western blotting demonstrated that treatment with TS and GA suppressed the activity or expression of MMP-9, MMP-2, and t-PA. Additionally, TS and GA significantly inhibited LPS-induced VEGF, PDGF, and VCAM-1 expression. Further investigation revealed that the inhibition of iNOS/COX-2, MMPs, growth factors, and adhesion molecules was associated with the suppression of NF-?B activation and MAPK (ERK1/2, JNK1/2, and p38) phosphorylation. Thus, Toona sinensis may be useful for the prevention of atherosclerosis. PMID:24723997

Yang, Hsin-Ling; Huang, Pei-Jane; Liu, Yi-Ru; Kumar, K. J. Senthil; Hsu, Li-Sung; Lu, Te-Ling; Chia, Yi-Chen; Takajo, Tokuko; Kazunori, Anzai; Hseu, You-Cheng

2014-01-01

117

BQ-123 prevents LPS-induced preterm birth in mice via the induction of uterine and placental IL-10.  

PubMed

Preterm birth (PTB), defined as any delivery occurring prior to the completion of 37 weeks' gestation, currently accounts for 11-12% of all births in the United States. Maternal genito-urinary infections account for up to 40% of all PTBS and induce a pro-inflammatory state in the host. The potent vasoconstrictor Endothelin-1 (ET-1) is known to be upregulated in the setting of infection, and elicits its effect by binding to the ETA receptor. We have previously shown that antagonism of the ETA receptor with BQ-123 is capable of preventing LPS-induced PTB in mice. We hypothesize that the administration of BQ-123 post LPS exposure will dismantle a positive feedback loop observed with pro-inflammatory cytokines upstream of ET-1. On GD 15.5, pregnant C57BL/6 mice were injected with PBS, LPS, BQ-123, or LPS+BQ-123. Changes at both the level of transcription and translation were observed in uterus and placenta in the ET-1 axis and in pro- and anti-inflammatory cytokines over the course of 12h. We discovered that BQ-123, when administered 10h post LPS, is capable of increasing production of uterine and placental Interleukin-10, causing a shift away from the pro-inflammatory state. We also observed that antagonism of the ETA receptor decreased IL-1? and TNF? in the placenta while also decreasing transcription of ET-1 in the uterus. Our results reinforce the role of ET-1 at the maternal fetal interface and highlight the potential benefit of ETA receptor blockade via the suppression of ET-1, and induction of a Th2 cytokine dominant state. PMID:25230003

Olgun, Nicole S; Hanna, Nazeeh; Reznik, Sandra E

2015-02-01

118

Regulatory role of C5a in LPS-induced IL-6 production by neutrophils during sepsis.  

PubMed

Experimental sepsis in rodents occurring after cecal ligation/puncture (CLP) is associated with excessive complement activation and a systemic inflammatory response. The proinflammatory mediator IL-6 has recently been shown to be an important inducer of the C5a receptor (C5aR) during sepsis. We now provide evidence that serum IL-6 production during sepsis in rats was reduced in neutrophil-depleted animals and that absence of C5aR in mice as well as antibody-blockade of C5a in rats significantly reduced serum levels of IL-6 during sepsis. Lipopolysaccharide (LPS)-induced production in vitro of IL-6 by neutrophils was significantly enhanced in the co-presence of C5a, likely due to transcriptional up-regulation of IL-6. Production of IL-6 in neutrophils by LPS was NF-kappaB dependent (but not on the presence of p50) and dependent on phosphorylation of p38-mitogen activated protein kinase (MAPK) as well as p44/p42 MAPK (ERK1/2) but not on phosphorylation of c-Jun N-terminal kinases (JNK1/2). C5a stimulation of neutrophils elicited a rapid phosphorylation of ERK1/2 and p38 MAPK. Accordingly, we suggest that induction of IL-6 after CLP is neutrophil and C5a/C5aR dependent, likely due to the ability of C5a to cause activation of ERK1/2 and p38 MAPK signaling pathways. PMID:14688199

Riedemann, Niels C; Guo, Ren-Feng; Hollmann, Travis J; Gao, Hongwei; Neff, Thomas A; Reuben, Jayne S; Speyer, Cecilia L; Sarma, J Vidya; Wetsel, Rick A; Zetoune, Firas S; Ward, Peter A

2004-02-01

119

Gamma-irradiated resveratrol negatively regulates LPS-induced MAPK and NF-?B signaling through TLR4 in macrophages.  

PubMed

Resveratrol was irradiated at various doses of 15, 30, 50, and 70kGy for the development of physiological functionalities through modification of the structural properties. Gamma irradiation induced a decrease in the resveratrol peak, and the appearance of several new peaks by gamma irradiation was gradually increased up to 70kGy. Gamma-irradiated resveratrol did not exert cytotoxicity to macrophages in dose ranges from 15 to 70kGy; therefore, 70kGy gamma-irradiated resveratrol was used as the maximum dose throughout subsequent experiments. Treatment of LPS-stimulated macrophages with 70kGy gamma-irradiated resveratrol resulted in a dose-dependent decrease in iNOS-mediated NO, PGE2, and pro-inflammatory cytokine level, such as TNF-?, IL-6 and IL-1?. 70kGy gamma-irradiated resveratrol significantly inhibited cyclooxygenase-2 levels, as well as the expression of cell surface molecules, such as CD80 and CD86, in LPS-induced macrophages. Furthermore, the inhibitory action of these pro-inflammatory mediators occurred through an inhibition of MAPKs (ERK1/2, p38 and JNK) and NF-?B signaling pathways based on a toll-like receptor 4 in macrophages, which may be closely mediated with the radiolysis products of resveratrol transformed by gamma-irradiation. From these findings, it seems likely that gamma irradiation can be an effective tool for a reduction of the toxicity and play a potent role in the treatment of inflammatory disease. PMID:25701505

Byun, Eui-Baek; Sung, Nak-Yun; Park, Jae-Nam; Yang, Mi-So; Park, Sang-Hyun; Byun, Eui-Hong

2015-04-01

120

Effect of anti-dementia drugs on LPS induced neuroinflammation in mice  

Microsoft Academic Search

Inflammation has been recently implicated in pathogenesis of dementia disorders. Effect of anti-dementia (Acetylcholinesterase inhibitor) drugs tacrine, rivastigmine and donepezil were studied on neuroinflammation induced by intraperitoneal administration of lipopolysaccharide (LPS) in mice. Interleukin-2 (IL-2) and isoforms of acetylcholinesterase (AChE) were estimated in different brain areas as marker for neuroinflammation and cholinergic activity respectively. LPS significantly increased the level of

Ethika Tyagi; Rahul Agrawal; Chandishwar Nath; Rakesh Shukla

2007-01-01

121

Characterization of LPS-induced TNF? factor (LITAF) from orange-spotted grouper, Epinephelus coioides.  

PubMed

Lipopolysaccharide-induced TNF? factor (LITAF) is an important transcription factor that mediates cell apoptosis and inflammatory response. In the present study, we cloned and characterized a LITAF gene from orange-spotted grouper (Epinephelus coioides) (Ec-LITAF). Ec-LITAF encoded a predicted 142 amino acid protein which shared 74% identity to sablefish (Anoplopoma fimbria) LITAF homolog. Multiple amino acid alignment showed that Ec-LITAF contained a typical LITAF domain with two CXXC motifs. Phylogenetic analysis indicated that Ec-LITAF was closely related to that of sablefish. Ec-LITAF mRNA was widely expressed in different tissues and its expression level in spleen was up-regulated after Singapore grouper iridovirus (SGIV) infection. Subcellular localization analysis revealed that the distribution of Ec-LITAF showed diffuse and aggregated patterns in cytoplasm. Interestingly, the distribution of Ec-LITAF overlayed with a viral LITAF homolog (vLITAF) encoded by SGIV. Overexpression of Ec-LITAF in vitro up-regulated the expression of tumor necrosis factors (TNF1 and TNF2) and TNF receptors (TNFR1 and TNFR2), and the expression of itself initiated apoptosis in fish cells. In addition, overexpression of Ec-LITAF not only accelerated SGIV infection induced CPE and cell death, but also increased viral gene transcription. Taken together, our data suggested that Ec-LITAF might play crucial roles during SGIV replication. PMID:24091064

Cai, Jia; Huang, Youhua; Wei, Shina; Ouyang, Zhengliang; Huang, Xiaohong; Qin, Qiwei

2013-12-01

122

Preferential macrophage recruitment and polarization in LPS-induced animal model for COPD: noninvasive tracking using MRI.  

PubMed

Noninvasive imaging of macrophages activity has raised increasing interest for diagnosis of chronic obstructive respiratory diseases (COPD), which make them attractive vehicles to deliver contrast agents for diagnostic or drugs for therapeutic purposes. This study was designed to monitor and evaluate the migration of differently polarized M1 and M2 iron labeled macrophage subsets to the lung of a LPS-induced COPD animal model and to assess their polarization state once they have reached the inflammatory sites in the lung after intravenous injection. Ex vivo polarized bone marrow derived M1 or M2 macrophages were first efficiently and safely labeled with amine-modified PEGylated dextran-coated SPIO nanoparticles and without altering their polarization profile. Their biodistribution in abdominal organs and their homing to the site of inflammation in the lung was tracked for the first time using a free-breathing non-invasive MR imaging protocol on a 4.7T magnet after their intravenous administration. This imaging protocol was optimized to allow both detection of iron labeled macrophages and visualization of inflammation in the lung. M1 and M2 macrophages were successfully detected in the lung starting from 2 hours post injection with no variation in their migration profile. Quantification of cytokines release, analysis of surface membrane expression using flow cytometry and immunohistochemistry investigations confirmed the successful recruitment of injected iron labeled macrophages in the lung of COPD mice and revealed that even with a continuum switch in the polarization profile of M1 and M2 macrophages during the time course of inflammation a balanced number of macrophage subsets predominate. PMID:24598763

Al Faraj, Achraf; Sultana Shaik, Asma; Pureza, Mary Angeline; Alnafea, Mohammad; Halwani, Rabih

2014-01-01

123

Preferential Macrophage Recruitment and Polarization in LPS-Induced Animal Model for COPD: Noninvasive Tracking Using MRI  

PubMed Central

Noninvasive imaging of macrophages activity has raised increasing interest for diagnosis of chronic obstructive respiratory diseases (COPD), which make them attractive vehicles to deliver contrast agents for diagnostic or drugs for therapeutic purposes. This study was designed to monitor and evaluate the migration of differently polarized M1 and M2 iron labeled macrophage subsets to the lung of a LPS-induced COPD animal model and to assess their polarization state once they have reached the inflammatory sites in the lung after intravenous injection. Ex vivo polarized bone marrow derived M1 or M2 macrophages were first efficiently and safely labeled with amine-modified PEGylated dextran-coated SPIO nanoparticles and without altering their polarization profile. Their biodistribution in abdominal organs and their homing to the site of inflammation in the lung was tracked for the first time using a free-breathing non-invasive MR imaging protocol on a 4.7T magnet after their intravenous administration. This imaging protocol was optimized to allow both detection of iron labeled macrophages and visualization of inflammation in the lung. M1 and M2 macrophages were successfully detected in the lung starting from 2 hours post injection with no variation in their migration profile. Quantification of cytokines release, analysis of surface membrane expression using flow cytometry and immunohistochemistry investigations confirmed the successful recruitment of injected iron labeled macrophages in the lung of COPD mice and revealed that even with a continuum switch in the polarization profile of M1 and M2 macrophages during the time course of inflammation a balanced number of macrophage subsets predominate. PMID:24598763

Al Faraj, Achraf; Sultana Shaik, Asma; Pureza, Mary Angeline; Alnafea, Mohammad; Halwani, Rabih

2014-01-01

124

Genomic Instability in Liver Cells Caused by an LPS-Induced Bystander-Like Effect  

PubMed Central

Bacterial infection has been linked to carcinogenesis, however, there is lack of knowledge of molecular mechanisms that associate infection with the development of cancer. We analyzed possible effects of the consumption of heat-killed E. coli O157:H7 cells or its cellular components, DNA, RNA, protein or lipopolysaccharides (LPS) on gene expression in naïve liver cells. Four week old mice were provided water supplemented with whole heat-killed bacteria or bacterial components for a two week period. One group of animals was sacrificed immediately, whereas another group was allowed to consume uncontaminated tap water for an additional two weeks, and liver samples were collected, post mortem. Liver cells responded to exposure of whole heat-killed bacteria and LPS with alteration in ?H2AX levels and levels of proteins involved in proliferation, DNA methylation (MeCP2, DNMT1, DNMT3A and 3B) or DNA repair (APE1 and KU70) as well as with changes in the expression of genes involved in stress response, cell cycle control and bile acid biosynthesis. Other bacterial components analysed in this study did not lead to any significant changes in the tested molecular parameters. This study suggests that lipopolysaccharides are a major component of Gram-negative bacteria that induce molecular changes within naïve cells of the host. PMID:23874414

Kovalchuk, Igor; Walz, Paul; Thomas, James; Kovalchuk, Olga

2013-01-01

125

Curcumin modulates the immune response associated with LPS-induced periodontal disease in rats  

PubMed Central

Curcumin is a plant-derived dietary spice ascribed various biological activities. Curcumin therapeutic applications have been studied in a variety of conditions, but not on periodontal disease. Periodontal disease is a chronic inflammatory condition initiated by an immune response to microorganisms of the dental biofilm. Experimental periodontal disease was induced in rats by injecting LPS in the gingival tissues on the palatal aspect of upper first molars (30 ug LPS, 3 times/week for 2 weeks). Curcumin was administered to rats daily via oral gavage at 30 and 100 mg/Kg. RT-qPCR and ELISA were used to determine the expression of IL-6, TNF-? and PGE2 synthase on the gingival tissues. The inflammatory status was evaluated by stereometric and descriptive analysis on H&E-stained sections, whereas modulation of p38 MAPK and NK-?B signaling was assessed by Western blot. Curcumin effectively inhibited cytokine gene expression at mRNA and protein levels, but NF-kB was inhibited only with the lower dose of curcumin, whereas p38 MAPK activation was not affected. Curcumin produced a significant reduction on the inflammatory infiltrate and increased collagen content and fibroblastic cell numbers. Curcumin potently inhibits innate immune responses associated with periodontal disease, suggesting a therapeutic potential in this chronic inflammatory condition. PMID:21242275

Guimarães, Morgana R.; de Aquino, Sabrina Garcia; Coimbra, Leila S.; Spolidorio, Luis C.; Kirkwood, Keith L.; Rossa, Carlos

2011-01-01

126

Cloning and analysis of gene regulation of a novel LPS-inducible cDNA  

SciTech Connect

The expression of many genes is altered upon the activation of macrophages by bacterial LPS. These genes play a crucial role in the orchestration of various responses to protect the host against infection. A novel 2.3 kilobase (kb) cDNA, designated IRG1, was obtained from a cDNA library prepared with RNA isolated from RAW 264.7 following lipopolysaccharide stimulation. Sequence analysis of the clone revealed no identity to any known genes but showed the presence of many potential phosphorylation sites suggesting that IRG1 protein product may be regulated at this level. Furthermore, IRG1 contains the motif for glycosaminoglycan attachment site, implying that IRG1 may be a proteoglycan. By interspecific backcross analysis, IRG1 was mapped to mouse chromosome 14 linked to Tyrp2 and Rap2a. The IRG1 message appears 1.5 h following LPS exposure and its induction was not dependent on new protein synthesis. In fact, cycloheximide induced the expression of IRG1, suggesting that a protein repressor prevents the expression of IRG1 when uninduced. The role of the protein kinase A pathway in regulating the induction of IRG1 by LPS is questionable, because although forskolin inhibited its induction, neither dibutyrl-cAMP nor 8-(4-chlorophenylthio)-cAMP had much effect on its expression. In contrast, activation of protein kinase C potentiated the LPS response. Chelation of extracellular calcium inhibited IRG1 4 h after LPS induction, while increasing intracellular calcium had little effect on the levels of the IRG1 transcript. Inhibiting tyrosine phosphorylation abrogated the induction of IRG1 by LPS. Hence, the induction of IRG1 by LPS is mediated by tyrosine kinase and protein kinase C pathway. 80 refs., 5 figs.

Lee, C.G.L.; O`Brien, W.E. [Baylor College of Medicine, Houston, TX (United States); Jenkins, N.A.; Gilbert, D.J.; Copeland, N.G. [Frederick Cancer Research and Development Center, MD (United States)

1995-03-01

127

Epidural analgesia with morphine or buprenorphine in ponies with lipopolysaccharide (LPS)-induced carpal synovitis  

PubMed Central

This study evaluated the analgesia effects of the epidural administration of 0.1 mg/kg bodyweight (BW) of morphine or 5 ?g/kg BW of buprenorphine in ponies with radiocarpal joint synovitis. Six ponies were submitted to 3 epidural treatments: the control group (C) received 0.15 mL/kg BW of a 0.9% sodium chloride (NaCl) solution; group M was administered 0.1 mg/kg BW of morphine; and group B was administered 5 ?g/kg BW of buprenorphine, both diluted in 0.9% NaCl to a total volume of 0.15 mL/kg BW administered epidurally at 10 s/mL. The synovitis model was induced by injecting 0.5 ng of lipopolysaccharide (LPS) in the left or right radiocarpal joint. An epidural catheter was later introduced in the lumbosacral space and advanced up to the thoracolumbar level. The treatment started 6 h after synovitis induction. Lameness, maximum angle of carpal flexion, heart rate, systolic arterial pressure, respiratory rate, temperature, and intestinal motility were evaluated before LPS injection (baseline), 6 h after LPS injection (time 0), and 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, and 24 h after treatments. Although the model of synovitis produced clear clinical signs of inflammation, the lameness scores in group C were different from the baseline for only up to 12 h. Both morphine and buprenorphine showed a reduction in the degree of lameness starting at 0.5 and 6 h, respectively. Reduced intestinal motility was observed at 0.5 h in group M and at 0.5 to 1 h in group B. Epidural morphine was a more effective analgesic that lasted for more than 12 h and without side effects. It was concluded that morphine would be a valuable analgesic option to alleviate joint pain in the thoracic limbs in ponies. PMID:21731186

Freitas, Gabrielle C.; Carregaro, Adriano B.; Gehrcke, Martielo I.; De La Côrte, Flávio D.; Lara, Valéria M.; Pozzobon, Ricardo; Brass, Karin E.

2011-01-01

128

Down-regulation of MAPK/NF-?B signaling underlies anti-inflammatory response induced by transduced PEP-1-Prx2 proteins in LPS-induced Raw 264.7 and TPA-induced mouse ear edema model.  

PubMed

Excessive reactive oxygen species (ROS) production plays a crucial role in causing various diseases, including inflammatory disorders. The activation of mitogen-activated protein kinase (MAPK) and nuclear factor-kappaB (NF-?B) signaling is implicated in stimulating inflammatory response and cytokines. Peroxiredoxin 2 (Prx2) is a 2-cysteine (Cys) peroxiredoxin capable of removing endogenous hydrogen peroxide (H2O2). PEP-1 peptide, a protein transduction domain, consists of three domains which are used to transduce exogenous therapeutic proteins into cells. The correlation between effectively transduced PEP-1-Prx2 and ROS-mediated inflammatory response is not clear. In the present study, we investigated the protective effects of cell permeable PEP-1-Prx2 on oxidative stress-induced inflammatory activity in Raw 264.7 cells and in a mouse ear edema model after exposure to lipopolysaccharides (LPS) or 12-O-tetradecanoylphorbol-13-acetate (TPA). Transduced PEP-1-Prx2 suppressed intracellular ROS accumulation and inhibited the activity of MAPKs and NF-?B signaling that led to the suppression of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS) and cytokines in LPS-induced Raw 264.7 cells and TPA-induced mouse ear edema model. Given these results, we propose that PEP-1-Prx2 has therapeutic potential in the prevention of inflammatory disorders. PMID:25241246

Jeong, Hoon Jae; Park, Meeyoung; Kim, Dae Won; Ryu, Eun Ji; In Yong, Ji; Cha, Hyun Ju; Kim, Sang Jin; Yeo, Hyeon Ji; Jeong, Ji-Heon; Kim, Duk-Soo; Kim, Hyoung Chun; Shin, Eun Joo; Park, Eun Young; Park, Jong Hoon; Kwon, Hyeok Yil; Park, Jinseu; Eum, Won Sik; Choi, Soo Young

2014-12-01

129

IL1 ?- and LPS-induced serotonin secretion is increased in EC cells derived from Crohn's disease  

PubMed Central

Gut mucosal enterochromaffin (EC) cells are regarded as key regulators of intestinal motility and fluid secretion via secretion of serotonin (5HT), are increased in numbers in mucosal inflammation and located in close proximity to immune cells. We examined whether interleukin (IL)1? and Escherichia coli lipopolysaccharide (LPS) induced EC cell 5HT release through Toll-like/IL-1 (TIL) receptor activation, nuclear factor kappa B (NF?B) and mitogen-activated protein kinase (MAPK) phosphorylation and evaluated whether somatostatin could inhibit this phenomenon. Pure (>98%) human intestinal EC cells were isolated by fluorescent activated cell sorting from preparations of normal (n = 5) and Crohn's colitis (n = 6) mucosa. 5HT release was measured (ELISA), and NF?B and ERK phosphorylation quantitated (ELISA) in response to IL1? and LPS. 5HT secretion was increased by both E. coli LPS (EC50 = 5 ng mL?1) and IL1? (EC50 = 0.05 pmol L?1) >2-fold (P < 0.05) in Crohn's EC cells compared with normal EC cells. Secretion was reversible by the TLR4 antagonist, E. coli K12 LPS (IC50 = 12 ng mL?1) and the IL1? receptor antagonist (ILRA; IC50 = 3.4 ng mL?1). IL1? caused significant (P < 0.05) NF?B and MAPK phosphorylation (40–55%). The somatostatin analogue, lanreotide inhibited IL1b-stimulated secretion in Crohn's (IC50 = 0.61 nmol L?1) and normal EC cells (IC50 = 1.8 nmol L?1). Inter-leukins (IL1?) and bacterial products (E. coli LPS) stimulated 5HT secretion from Crohn's EC cells via TIL receptor activation (TLR4 and IL1?). Immune-mediated alterations in EC cell secretion of 5HT may represent a component of the pathogenesis of abnormal bowel function in Crohn's disease. Inhibition of EC cell-mediated 5HT secretion may be an alternative therapeutic strategy in the amelioration of inflammatory bowel disease symptomatology. PMID:19019013

KIDD, M.; GUSTAFSSON, B. I.; DROZDOV, I.; MODLIN, I. M.

2014-01-01

130

Gypenoside XLIX, a naturally occurring gynosaponin, PPAR-alpha dependently inhibits LPS-induced tissue factor expression and activity in human THP-1 monocytic cells  

SciTech Connect

Tissue factor (TF) is involved not only in the progression of atherosclerosis and other cardiovascular diseases, but is also associated with tumor growth, metastasis, and angiogenesis and hence may be an attractive target for directed cancer therapeutics. Gynostemma pentaphyllum (GP) is widely used in the treatment of various cardiovascular diseases including atherosclerosis, as well as cancers. Gypenoside (Gyp) XLIX, a dammarane-type glycoside, is one of the prominent components in GP. We have recently reported Gyp XLIX to be a potent peroxisome proliferator-activated receptor (PPAR)-alpha activator. Here we demonstrate that Gyp XLIX (0-300 {mu}M) concentration dependently inhibited TF promoter activity after induction by the inflammatory stimulus lipopolysaccharide (LPS) in human monocytic THP-1 cells transfected with promoter reporter constructs pTF-LUC. Furthermore, Gyp XLIX inhibited LPS-induced TF mRNA and protein overexpression in THP-1 monocyte cells. Its inhibition of LPS-induced TF hyperactivity was further confirmed by chromogenic enzyme activity assay. The activities of Gyp XLIX reported in this study were similar to those of Wy-14643, a potent synthetic PPAR-alpha activator. Furthermore, the Gyp XLIX-induced inhibitory effect on TF luciferase activity was completely abolished in the presence of the PPAR-alpha selective antagonist MK-886. The present findings suggest that Gyp XLIX inhibits LPS-induced TF overexpression and enhancement of its activity in human THP-1 monocytic cells via PPAR-alpha-dependent pathways. The data provide new insights into the basis of the use of the traditional Chinese herbal medicine G. pentaphyllum for the treatment of cardiovascular and inflammatory diseases, as well as cancers.

Huang, Tom Hsun-Wei [Faculty of Pharmacy, University of Sydney, NSW 2006 (Australia); Van Hoan Tran [Faculty of Pharmacy, University of Sydney, NSW 2006 (Australia); Roufogalis, Basil D. [Faculty of Pharmacy, University of Sydney, NSW 2006 (Australia); Li Yuhao [Faculty of Pharmacy, University of Sydney, NSW 2006 (Australia)]. E-mail: yuhao@pharm.usyd.edu.au

2007-01-01

131

GM-CSF increases LPS-induced production of proinflammatory mediators via upregulation of TLR4 and CD14 in murine microglia  

PubMed Central

Background Microglia are resident macrophage-like cells in the central nervous system (CNS) and cause innate immune responses via the LPS receptors, Toll-like receptor (TLR) 4 and CD14, in a variety of neuroinflammatory disorders including bacterial infection, Alzheimer’s disease, and amyotrophic lateral sclerosis. Granulocyte macrophage-colony stimulating factor (GM-CSF) activates microglia and induces inflammatory responses via binding to GM-CSF receptor complex composed of two different subunit GM-CSF receptor ? (GM-CSFR?) and common ? chain (?c). GM-CSF has been shown to be associated with neuroinflammatory responses in multiple sclerosis and Alzheimer’s disease. However, the mechanisms how GM-CSF promotes neuroinflammation still remain unclear. Methods Microglia were stimulated with 20 ng/ml GM-CSF and the levels of TLR4 and CD14 expression were evaluated by RT-PCR and flowcytometry. LPS binding was analyzed by flowcytometry. GM-CSF receptor complex was analyzed by immunocytechemistry. The levels of IL-1?, IL-6 and TNF-? in culture supernatant of GM-CSF-stimulated microglia and NF-?B nuclear translocation were determined by ELISA. Production of nitric oxide (NO) was measured by the Griess method. The levels of p-ERK1/2, ERK1/2, p-p38 and p38 were assessed by Western blotting. Statistically significant differences between experimental groups were determined by one-way ANOVA followed by Tukey test for multiple comparisons. Results GM-CSF receptor complex was expressed in microglia. GM-CSF enhanced TLR4 and CD14 expressions in microglia and subsequent LPS-binding to the cell surface. In addition, GM-CSF priming increased LPS-induced NF-?B nuclear translocation and production of IL-1?, IL-6, TNF-? and NO by microglia. GM-CSF upregulated the levels of p-ERK1/2 and p-p38, suggesting that induction of TLR4 and CD14 expression by GM-CSF was mediated through ERK1/2 and p38, respectively. Conclusions These results suggest that GM-CSF upregulates TLR4 and CD14 expression in microglia through ERK1/2 and p38, respectively, and thus promotes the LPS receptor-mediated inflammation in the CNS. PMID:23234315

2012-01-01

132

Curcumin inhibits LPS-induced inflammation in rat vascular smooth muscle cells in vitro via ROS-relative TLR4-MAPK/NF-?B pathways  

PubMed Central

Aim: To investigate whether curcumin (Cur) suppressed lipopolysaccharide (LPS)-induced inflammation in vascular smooth muscle cells (VSMCs) of rats, and to determine its molecular mechanisms. Methods: Primary rat VSMCs were treated with LPS (1 ?g/L) and Cur (5, 10, or 30 ?mol/L) for 24 h. The levels of MCP-1, TNF-?, and iNOS were measured using ELISA and real-time RT-PCR. NO level was analyzed with the Griess reaction. Western-blotting was used to detect the activation of TLR4, MAPKs, I?B?, NF-?B p65, and the p47phox subunit of NADPH oxidase in the cells. Results: Treatment of VSMCs with LPS dramatically increased expression of inflammatory cytokines MCP-1 and TNF-?, expression of TLR4 and iNOS, and NO production. LPS also significantly increased phosphorylation of I?B?, nuclear translocation of NF-?B (p65) and phosphorylation of MAPKs in VSMCs. Furthermore, LPS significantly increased production of intracellular ROS, and decreased expression of p47phox subunit of NADPH oxidase. Pretreatment with Cur concentration-dependently attenuated all the aberrant changes in LPS-treated VSMCs. The LPS-induced overexpression of MCP-1 and TNF-?, and NO production were attenuated by pretreatment with the ERK inhibitor PD98059, the p38 MAPK inhibitor SB203580, the NF-?B inhibitor PDTC or anti-TLR4 antibody, but not with the JNK inhibitor SP600125. Conclusion: Cur suppresses LPS-induced overexpression of inflammatory mediators in VSMCs in vitro via inhibiting the TLR4-MAPK/NF-?B pathways, partly due to block of NADPH-mediated intracellular ROS production. PMID:23645013

Meng, Zhe; Yan, Chao; Deng, Qian; Gao, Deng-feng; Niu, Xiao-lin

2013-01-01

133

Vitamin C Mitigates Oxidative Stress and Tumor Necrosis Factor-Alpha in Severe Community-Acquired Pneumonia and LPS-Induced Macrophages  

PubMed Central

Oxidative stress is an important part of host innate immune response to foreign pathogens. However, the impact of vitamin C on oxidative stress and inflammation remains unclear in community-acquired pneumonia (CAP). We aimed to determine the effect of vitamin C on oxidative stress and inflammation. CAP patients were enrolled. Reactive oxygen species (ROS), DNA damage, superoxide dismutases (SOD) activity, tumor necrosis factor-alpha (TNF-?), and IL-6 were analyzed in CAP patients and LPS-stimulated macrophages cells. MH-S cells were transfected with RFP-LC3 plasmids. Autophagy was measured in LPS-stimulated macrophages cells. Severe CAP patients showed significantly increased ROS, DNA damage, TNF-?, and IL-6. SOD was significantly decreased in severe CAP. Vitamin C significantly decreased ROS, DNA damage, TNF-?, and IL-6. Vitamin C inhibited LPS-induced ROS, DNA damage, TNF-?, IL-6, and p38 in macrophages cells. Vitamin C inhibited autophagy in LPS-induced macrophages cells. These findings indicated that severe CAP exhibited significantly increased oxidative stress, DNA damage, and proinflammatory mediator. Vitamin C mitigated oxidative stress and proinflammatory mediator suggesting a possible mechanism for vitamin C in severe CAP. PMID:25253919

Chen, Yuanyuan; Luo, Guangyan; Yuan, Jiao; Wang, Yuanyuan; Yang, Xiaoqiong; Wang, Xiaoyun; Li, Guoping; Liu, Zhiguang; Zhong, Nanshan

2014-01-01

134

Pre-Treatment of Recombinant Mouse MFG-E8 Downregulates LPS-Induced TNF-? Production in Macrophages via STAT3-Mediated SOCS3 Activation  

PubMed Central

Milk fat globule-epidermal growth factor factor 8 (MFG-E8) regulates innate immune function by modulating cellular signaling, which is less understood. Herein, we aimed to investigate the direct anti-inflammatory role of MFG-E8 in macrophages by pre-treatment with recombinant murine MFG-E8 (rmMFG-E8) followed by stimulation with LPS in RAW264.7 cells and in peritoneal macrophages, isolated from wild-type (WT) or MFG-E8?/? mice. RAW264.7 cells and mouse peritoneal macrophages treated with rmMFG-E8 significantly downregulated LPS-induced TNF-? mRNA by 25% and 24%, and protein levels by 29% and 23%, respectively (P<0.05). Conversely, peritoneal macrophages isolated from MFG-E8?/? mice produced 28% higher levels of TNF-?, as compared to WT mice when treated with LPS. In in vivo, endotoxemia induced by intraperitoneal injection of LPS (5 mg/kg BW), at 4 h after induction, serum level of TNF-? was significantly higher in MFG-E8?/? mice (837 pg/mL) than that of WT (570 pg/mL, P<0.05). To elucidate the direct anti-inflammatory effect of MFG-E8, we examined STAT3 and its target gene, SOCS3. Treatment with rmMGF-E8 significantly induced pSTAT3 and SOCS3 in macrophages. Similar results were observed in in vivo treatment of rmMFG-E8 in peritoneal cells and splenic tissues. Pre-treatment with rmMFG-E8 significantly reduced LPS-induced NF-?B p65 contents. These data clearly indicated that rmMFG-E8 upregulated SOCS3 which in turn interacted with NF-?B p65, facilitating negative regulation of TLR4 signaling for LPS-induced TNF-? production. Our findings strongly suggest that MFG-E8 is a direct anti-inflammatory molecule, and that it could be developed as a therapy in attenuating inflammation and tissue injury. PMID:22114683

Aziz, Monowar; Jacob, Asha; Matsuda, Akihisa; Wu, Rongqian; Zhou, Mian; Dong, Weifeng; Yang, Weng-Lang; Wang, Ping

2011-01-01

135

Quince (Cydonia oblonga Miller) peel polyphenols modulate LPS-induced inflammation in human THP-1-derived macrophages through NF-{kappa}B, p38MAPK and Akt inhibition  

SciTech Connect

Highlights: Black-Right-Pointing-Pointer Quince peel polyphenols inhibit LPS-induced secretion of TNF-{alpha} and IL-8. Black-Right-Pointing-Pointer Quince peel polyphenols augment LPS-induced secretion of IL-10 and IL-6. Black-Right-Pointing-Pointer Quince peel polyphenols-mediated inhibition of LPS-induced secretion of TNF-{alpha} is partially mediated by IL-6. Black-Right-Pointing-Pointer The anti-inflammatory effects of quince polyphenols pass through NF-{kappa}B, p38MAPK and Akt inhibition. -- Abstract: Chronic inflammation is a hallmark of several pathologies, such as rheumatoid arthritis, gastritis, inflammatory bowel disease, atherosclerosis and cancer. A wide range of anti-inflammatory chemicals have been used to treat such diseases while presenting high toxicity and numerous side effects. Here, we report the anti-inflammatory effect of a non-toxic, cost-effective natural agent, polyphenolic extract from the Tunisian quince Cydonia oblonga Miller. Lipopolysaccharide (LPS) treatment of human THP-1-derived macrophages induced the secretion of high levels of the pro-inflammatory cytokine TNF-{alpha} and the chemokine IL-8, which was inhibited by quince peel polyphenolic extract in a dose-dependent manner. Concomitantly, quince polyphenols enhanced the level of the anti-inflammatory cytokine IL-10 secreted by LPS-treated macrophages. We further demonstrated that the unexpected increase in IL-6 secretion that occurred when quince polyphenols were associated with LPS treatment was partially responsible for the polyphenols-mediated inhibition of TNF-{alpha} secretion. Biochemical analysis showed that quince polyphenols extract inhibited the LPS-mediated activation of three major cellular pro-inflammatory effectors, nuclear factor-kappa B (NF-{kappa}B), p38MAPK and Akt. Overall, our data indicate that quince peel polyphenolic extract induces a potent anti-inflammatory effect that may prove useful for the treatment of inflammatory diseases and that a quince-rich regimen may help to prevent and improve the treatment of such diseases.

Essafi-Benkhadir, Khadija [Laboratoire d'epidemiologie Moleculaire et Pathologie Experimentale Appliquee Aux Maladies Infectieuses, Institut Pasteur de Tunis (Tunisia)] [Laboratoire d'epidemiologie Moleculaire et Pathologie Experimentale Appliquee Aux Maladies Infectieuses, Institut Pasteur de Tunis (Tunisia); Refai, Amira [Laboratoire de Recherche sur la Transmission, le Controle et l'immunobiologie des Infections, Institut Pasteur de Tunis (Tunisia)] [Laboratoire de Recherche sur la Transmission, le Controle et l'immunobiologie des Infections, Institut Pasteur de Tunis (Tunisia); Riahi, Ichrak [Laboratoire d'epidemiologie Moleculaire et Pathologie Experimentale Appliquee Aux Maladies Infectieuses, Institut Pasteur de Tunis (Tunisia)] [Laboratoire d'epidemiologie Moleculaire et Pathologie Experimentale Appliquee Aux Maladies Infectieuses, Institut Pasteur de Tunis (Tunisia); Fattouch, Sami [Laboratory LIP-MB National Institute of Applied Sciences and Technology, Tunis (Tunisia)] [Laboratory LIP-MB National Institute of Applied Sciences and Technology, Tunis (Tunisia); Karoui, Habib [Laboratoire d'epidemiologie Moleculaire et Pathologie Experimentale Appliquee Aux Maladies Infectieuses, Institut Pasteur de Tunis (Tunisia)] [Laboratoire d'epidemiologie Moleculaire et Pathologie Experimentale Appliquee Aux Maladies Infectieuses, Institut Pasteur de Tunis (Tunisia); Essafi, Makram, E-mail: makram.essafi@pasteur.rns.tn [Laboratoire de Recherche sur la Transmission, le Controle et l'immunobiologie des Infections, Institut Pasteur de Tunis (Tunisia)] [Laboratoire de Recherche sur la Transmission, le Controle et l'immunobiologie des Infections, Institut Pasteur de Tunis (Tunisia)

2012-02-03

136

The inhibitory effects of Geranium thunbergii on interferon-?- and LPS-induced inflammatory responses are mediated by Nrf2 activation  

PubMed Central

Geranium thunbergii Sieb. et Zucc. (GT; which belongs to the Geraniaceae family) has been used as a traditional medicine in East Asia for the treatment of inflammatory diseases, including arthritis and diarrhea. However, the underlying mechanisms of the anti-inflammatory effects of GT remain poorly understood. In the present study, we examined the mechanisms responsible for the anti-inflammatory activity of GT in macrophages. The results revealed that GT significantly inhibited the lipopolysaccharide (LPS)- and interferon-? (IFN-?)-induced expression of pro-inflammatory genes, such as inducible nitric oxide synthase, tumor necrosis factor-? and interleukin-1?, as shown by RT-PCR. However, the inhibitory effects of GT on LPS- and IFN-?-induced inflammation were associated with an enhanced nuclear factor erythroid 2-related factor 2 (Nrf2) activity, but not with the suppression of nuclear factor (NF)-?B activity, as shown by western blot analysis. In addition, in bone marrow-derived macrophages (BMDM) isolated from Nrf2 knockout mice, GT did not exert any inhibitory effect on the LPS- and IFN-?-induced inflammation. Taken together, our findings indicate that the anti-inflammatory effects of GT may be associated with the activation of Nrf2, an anti-inflammatory transcription factor. PMID:25761198

CHOI, HEE-JIN; CHOI, HEE-JUNG; PARK, MI-JU; LEE, JI-YEON; JEONG, SEUNG-IL; LEE, SEONGOO; KIM, KYUN HA; JOO, MYUNGSOO; JEONG, HAN-SOL; KIM, JAI-EUN; HA, KI-TAE

2015-01-01

137

The inhibitory effects of Geranium thunbergii on interferon-?- and LPS-induced inflammatory responses are mediated by Nrf2 activation.  

PubMed

Geranium thunbergii Sieb. et Zucc. (GT; which belongs to the Geraniaceae family) has been used as a traditional medicine in East Asia for the treatment of inflammatory diseases, including arthritis and diarrhea. However, the underlying mechanisms of the anti-inflammatory effects of GT remain poorly understood. In the present study, we examined the mechanisms responsible for the anti-inflammatory activity of GT in macrophages. The results revealed that GT significantly inhibited the lipopolysaccharide (LPS)- and interferon-? (IFN-?)-induced expression of pro-inflammatory genes, such as inducible nitric oxide synthase, tumor necrosis factor-? and interleukin-1?, as shown by RT-PCR. However, the inhibitory effects of GT on LPS- and IFN-?-induced inflammation were associated with an enhanced nuclear factor erythroid 2-related factor 2 (Nrf2) activity, but not with the suppression of nuclear factor (NF)-?B activity, as shown by western blot analysis. In addition, in bone marrow-derived macrophages (BMDM) isolated from Nrf2 knockout mice, GT did not exert any inhibitory effect on the LPS- and IFN-?-induced inflammation. Taken together, our findings indicate that the anti-inflammatory effects of GT may be associated with the activation of Nrf2, an anti-inflammatory transcription factor. PMID:25761198

Choi, Hee-Jin; Choi, Hee-Jung; Park, Mi-Ju; Lee, Ji-Yeon; Jeong, Seung-Il; Lee, Seongoo; Kim, Kyun Ha; Joo, Myungsoo; Jeong, Han-Sol; Kim, Jai-Eun; Ha, Ki-Tae

2015-05-01

138

LPS-induced TNF? factor (LITAF) in the snail Cipangopaludina chinensis: gene cloning and its apoptotic effect on NCI-H446 cells.  

PubMed

LPS-induced TNF? factor (LITAF) is a transcription factor mediating TNF-? expression under LPS stimulation, and playing important roles in immune responses. In the present study, partial cDNA sequence of a LITAF (designated CcLITAF) gene was cloned and identified from snail Cipangopaludina chinensis. It contains an open reading frame of 348 nucleotides encoding a predicted protein of 115 amino acids, with a conserved LITAF domain at C-terminal, and shares a similarity ranging from 34% to 96% with other LITAF from oyster to mammals. CcLITAF mRNA ubiquitously expressed in all analyzed tissues. Interestingly, cLITAF could induce apoptosis in human tumor cell line, NCI-H446 cells, and caspase 3 play key roles in CcLITAF-mediated apoptosis. Present studies provide new insight into the biological function of CcLITAF. PMID:22138218

Yang, Shoubao; Li, Peifen; Mi, Zhongxiang

2012-02-01

139

PF-04886847 (an inhibitor of plasma kallikrein) attenuates inflammatory mediators and activation of blood coagulation in rat model of lipopolysaccharide (LPS)-induced sepsis.  

PubMed

The plasma kallikrein-mediated proteolysis regulates both thrombosis and inflammation. Previous study has shown that PF-04886847 is a potent and competitive inhibitor of kallikrein, suggesting that it might be useful for the treatment of kallikrein-kinin mediated inflammatory and thrombotic disorders. In the rat model of lipopolysaccharide (LPS) -induced sepsis used in this study, pretreatment of rats with PF-04886847 (1 mg/kg) prior to LPS (10 mg/kg) prevented endotoxin-induced increase in granulocyte count in the systemic circulation. PF-04886847 significantly reduced the elevated plasma 6-keto PGF1? levels in LPS treated rats, suggesting that PF-04886847 could be useful in preventing hypotensive shock during sepsis. PF-04886847 did not inhibit LPS-induced increase in plasma TNF-? level. Pretreatment of rats with PF-04886847 prior to LPS did not attenuate endotoxin-induced decrease in platelet count and plasma fibrinogen levels as well as increase in plasma D-dimer levels. PF-04886847 did not protect the animals against LPS-mediated acute hepatic and renal injury and disseminated intravascular coagulation (DIC). Since prekallikrein (the zymogen form of plasma kallikrein) deficient patients have prolonged activated partial thromboplastin time (aPTT) without having any bleeding disorder, the anti-thrombotic property and mechanism of action of PF-04886847 was assessed. In a rabbit balloon injury model designed to mimic clinical conditions of acute thrombotic events, PF-04886847 reduced thrombus mass dose-dependently. PF-04886847 (1 mg/kg) prolonged both aPTT and prothrombin time (PT) in a dose-dependent manner. Although the findings of this study indicate that PF-04886847 possesses limited anti-thrombotic and anti-inflammatory effects, PF-04886847 may have therapeutic potential in other kallikrein-kinin mediated diseases. PMID:22352684

Kolte, D; Bryant, J W; Gibson, G W; Wang, J; Shariat-Madar, Z

2012-06-01

140

Constitutive and LPS-Induced Expression of MCP-1 and IL-8 by Human Uveal Melanocytes In Vitro and Relevant Signal Pathways  

PubMed Central

Purpose. Melanocytes are one of the major cellular components in the uvea. Interleukin-8/CXCL8 and monocyte chemoattractant protein-1 (MCP-1/CCL2) are the two most important proinflammatory chemokines. We studied the constitutive and lipopolysaccharide (LPS)-induced expression of IL-8 and MCP-1 in cultured human uveal melanocytes (UM) and explored the relevant signal pathways. Methods. Conditioned media and cells were collected from UM cultured in medium with and without stimulation of LPS. Interleukin-8 and MCP-1 proteins and mRNAs were measured using an ELISA kit and RT-PCR, respectively. Nuclear factor (NF)-?B in nuclear extracts and phosphorylated p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinases1/2 (ERK1/2), and c-Jun N-terminal kinase1/2 (JNK1/2) in cells cultured with and without LPS were measured by ELISA kits. Inhibitors of p38 (SB203580), ERK1/2 (UO1026), JNK1/2 (SP600125), and NF-?B (BAY11-7082) were added to the cultures to evaluate their effects. Results. Low levels of IL-8 and MCP-1 proteins were detected in the conditioned media in UM cultured without serum. Lipopolysaccharide (0.01–1 ?g/mL) increased IL-8 and MCP-1 mRNAs and proteins levels in a dose- and time-dependent manner, accompanied by a significant increase of phosphorylated JNK1/2 in cell lysates and NF-?B in nuclear extracts. Nuclear factor–?B and JNK1/2 inhibitors significantly blocked LPS-induced expression of IL-8 and MCP-1. Conclusions. This is the first report on the expression and secretion of chemokines by UM. The data suggest that UM may play a role in the pathogenesis of ocular inflammatory diseases. PMID:25125602

Hu, Dan-Ning; Bi, Mingchao; Zhang, David Y.; Ye, Fei; McCormick, Steven A.; Chan, Chi-Chao

2014-01-01

141

Qing Hua Chang Yin inhibits the LPS-induced activation of the IL-6/STAT3 signaling pathway in human intestinal Caco-2 cells.  

PubMed

Increasing evidence indicates that the pathogenesis of ulcerative colitis (UC) is highly regulated by the interleukin-6 (IL-6)/signal transducer and activator of transcription 3 (STAT3) pathway and its negative feedback regulator, suppressor of cytokine signaling 3 (SOCS3). Therefore, modulating the signaling feedback loop of IL-6/STAT3/SOCS3 may prove to be a novel therapeutic approach for the treatment of UC. Qing Hua Chang Yin (QHCY) is a traditional Chinese formulation that has long been used in clinic for the treatment of UC. We have previously reported that QHCY ameliorates acute intestinal inflammation in vivo and in vitro through the suppression of the nuclear factor??B (NF-?B) pathway. In the present study, in order to further elucidate the mechanisms responsible for the anti?inflammatory activities of QHCY, we stimulated human intestinal Caco-2 cells with lipopolysaccharide (LPS) to create an in vitro model of an inflamed human intestinal epithelium, and evaluated the effects of QHCY on the IL-6/STAT3/SOCS3 signaling network in inflamed Caco-2 cells. The levels of IL-6 were measured by ELISA and the levels of STAT3 and SOCS3 were measured by western blot analysis. We found that QHCY significantly inhibited the LPS-induced secretion of pro-inflammatory IL-6 in the Caco-2 cells in a dose-dependent manner. Moreover, QHCY profoundly suppressed the LPS-induced phosphorylation of Janus-activated kinase 1 (JAK1), JAK2 and STAT3. Furthermore, treatment with QHCY markedly augmented the expression of SOCS3. Taken together, the findings of the present study suggest that the modulation of the IL-6/STAT3/SOCS3 signaling network may be one of the mechanisms through which QHCY exerts its anti?inflammatory effects. PMID:25633437

Ke, Xiao; Hu, Guanghong; Fang, Wenyi; Chen, Jintuan; Zhang, Xin; Yang, Chunbo; Peng, Jun; Chen, Youqin; Sferra, Thomas J

2015-04-01

142

Propofol pretreatment attenuates LPS-induced granulocyte-macrophage colony-stimulating factor production in cultured hepatocytes by suppressing MAPK/ERK activity and NF-{kappa}B translocation  

SciTech Connect

Propofol (PPF), a widely used intravenous anesthetic for induction and maintenance of anesthesia during surgeries, was found to possess suppressive effect on host immunity. This study aimed at investigating whether PPF plays a modulatory role in the lipopolysaccharide (LPS)-induced inflammatory cytokine expression in a cell line of rat hepatocytes. Morphological observation and viability assay showed that PPF exhibits no cytotoxicity at concentrations up to 300 {mu}M after 48 h incubation. Pretreatment with 100 {mu}M PPF for 24 h prior to LPS stimulation was performed to investigate the modulatory effect on LPS-induced inflammatory gene production. The results of semi-quantitative RT-PCR demonstrated that PPF pretreatment significantly suppressed the LPS-induced toll-like receptor (TLR)-4, CD14, tumor necrosis factor (TNF)-{alpha}, and granulocyte-macrophage colony-stimulating factor (GM-CSF) gene expression. Western blotting analysis showed that PPF pretreatment potentiated the LPS-induced TLR-4 downregulation. Flow cytometrical analysis revealed that PPF pretreatment showed no modulatory effect on the LPS-upregulated CD14 expression on hepatocytes. In addition, PPF pretreatment attenuated the phosphorylation of mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) and I{kappa}B{alpha}, as well as the nuclear translocation of NF-{kappa}B primed by LPS. Moreover, addition of PD98059, a MAPK kinase inhibitor, significantly suppressed the LPS-induced NF-{kappa}B nuclear translocation and GM-CSF production, suggesting that the PPF-attenuated GM-CSF production in hepatocytes may be attributed to its suppressive effect on MAPK/ERK signaling pathway. In conclusion, PPF as an anesthetic may clinically benefit those patients who are vulnerable to sepsis by alleviating sepsis-related inflammatory response in livers.

Jawan, Bruno [Department of Anesthesiology and Liver Transplantation Program, Chang Gung Memorial Hospital, Kaohsiung Medical Center, Chang Gung University College of Medicine, 123 Ta-Pei Road, Niao-Sung Hsiang, Kaohsiung Hsien 833, Taiwan (China); Kao, Y.-H. [Department of Anesthesiology and Liver Transplantation Program, Chang Gung Memorial Hospital, Kaohsiung Medical Center, Chang Gung University College of Medicine, 123 Ta-Pei Road, Niao-Sung Hsiang, Kaohsiung Hsien 833, Taiwan (China); Department of Biological Sciences, National Sun Yat-Sen University, 70 Lien-Hai Road, Kaohsiung 804, Taiwan (China); Goto, Shigeru [Department of Surgery and Liver Transplantation Program, Chang Gung Memorial Hospital, Kaohsiung Medical Center, Chang Gung University College of Medicine, 123 Ta-Pei Road, Niao-Sung Hsiang, Kaohsiung Hsien 833, Taiwan (China); Department of Surgery, Iwao Hospital, 3059-1 Kawakami, Yufuin, Oita 879-5102 (Japan); Pan, M.-C. [Department of Anesthesiology and Liver Transplantation Program, Chang Gung Memorial Hospital, Kaohsiung Medical Center, Chang Gung University College of Medicine, 123 Ta-Pei Road, Niao-Sung Hsiang, Kaohsiung Hsien 833, Taiwan (China); Lin, Y.-C.; Hsu, L.-W.; Nakano, Toshiaki; Lai, C.-Y.; Sun, C.-K.; Cheng, Y.-F. [Department of Surgery and Liver Transplantation Program, Chang Gung Memorial Hospital, Kaohsiung Medical Center, Chang Gung University College of Medicine, 123 Ta-Pei Road, Niao-Sung Hsiang, Kaohsiung Hsien 833, Taiwan (China); Tai, M.-H. [Department of Biological Sciences, National Sun Yat-Sen University, 70 Lien-Hai Road, Kaohsiung 804, Taiwan (China); Department of Medical Research and Education, Kaohsiung Veterans General Hospital, 386 Ta-Chung 1st Road, Kaohsiung 813, Taiwan (China)] (and others)

2008-06-15

143

SOCS3 Expression Correlates with Severity of Inflammation, Expression of Proinflammatory Cytokines, and Activation of STAT3 and p38 MAPK in LPS-Induced Inflammation In Vivo  

PubMed Central

SOCS3 is an inducible endogenous negative regulator of JAK/STAT pathway, which is relevant in inflammatory conditions. We used a model of LPS-induced periodontal disease in rats to correlate SOCS3 expression with the inflammatory status. In vitro we used a murine macrophage cell line to assess the physical interaction between SOCS3 and STAT3 by coimmunoprecipitation. 30?ug of LPS from Escherichia coli were injected in the gingival tissues on the palatal aspect of first molars of the animals 3x/week for up to 4 weeks. Control animals were injected with the vehicle (PBS). The rats were sacrificed at 7, 15, and 30 days. Inflammation and gene expression were assessed by stereometric analysis, immunohistochemistry, RT-qPCR, and western blot. LPS injections increased inflammation, paralleled by an upregulation of SOCS3, of the proinflammatory cytokines IL-1?, IL-6, and TNF-? and increased phosphorylation of STAT3 and p38 MAPK. SOCS3 expression accompanied the severity of inflammation and the expression of proinflammatory cytokines, as well as the activation status of STAT3 and p38 MAPK. LPS stimulation in a macrophage cell line in vitro induced transient STAT3 activation, which was inversely correlated with a dynamic physical interaction with SOCS3, suggesting that this may be a mechanism for SOCS3 regulatory function. PMID:24078776

Chaves de Souza, João Antônio; Nogueira, Andressa Vilas Boas; Chaves de Souza, Pedro Paulo; Kim, Yeon Jung; Silva Lobo, Caroline; Pimentel Lopes de Oliveira, Guilherme José; Cirelli, Joni Augusto; Garlet, Gustavo Pompermaier; Rossa, Carlos

2013-01-01

144

Anethole, a Medicinal Plant Compound, Decreases the Production of Pro-Inflammatory TNF-? and IL-1? in a Rat Model of LPS-Induced Periodontitis  

PubMed Central

Periodontitis (PD) is known to be one of most prevalent worldwide chronic inflammatory diseases. There are several treatments including antibiotics for PD; however, since drug resistance is an increasing problem, new drugs particularly derived from plants with fewer side effects are required. The effects of trans-anethole on IL-1 ? and TNF-? level in a rat model of PD were investigated and compared to ketoprofen. Eschericia coli lipopolysaccharide (LPS, 30 µg) was injected bilaterally into the palatal gingiva (3 µL/site) between the upper first and second molars every two days for 10 days in anesthetized rats. Administration of either trans-anethole (10 or 50 mg/Kg, i.p.) or ketoprofen (10 mg/Kg, i.p.) was started 20 minute before LPS injection and continued for 10 days. Then, IL-1? and TNF-? levels were measured in blood samples by ELISA at day 0 (control) and at day 10. Anethole at both concentrations significantly suppressed IL-1? and TNF-? production when compared to LPS-treated rats. The suppressive effects of anethole on LPS-induced pro-inflammatory cytokines were almost similar as seen with ketoprofen. In conclusion, the present results suggest that anethole may have a potent inhibitory effect on PD through suppression of pro-inflammatory molecules; therefore it could be a novel therapeutic strategy for PD. PMID:25587321

Moradi, Janet; Abbasipour, Fatemeh; Zaringhalam, Jalal; Maleki, Bita; Ziaee, Narges; Khodadoustan, Amin; Janahmadi, Mahyar

2014-01-01

145

LPS-induced NO inhibition and antioxidant activities of ethanol extracts and their solvent partitioned fractions from four brown seaweeds  

NASA Astrophysics Data System (ADS)

The nitric oxide inhibitory (NOI) and antioxidant (ABTS and DPPH radical scavenging effects with reducing power) activities of the ethanol (EtOH) extracts and solvent partitioned fractions from Scytosiphon lomentaria, Chorda filum, Agarum cribrosum, and Desmarestia viridis were investigated, and the correlation between biological activity and total phenolic (TP) and phlorotannin (TPT) content was determined by PCA analysis. The yield of EtOH extracts from four brown seaweeds ranged from 2.6 to 6.6% with the highest yield from D. viridis, and the predominant compounds in their solvent partitioned fractions had medium and/or less polarity. The TP and TPT content of the EtOH extracts were in the ranges of 25.0-44.1 mg GAE/g sample and 0.2-4.6 mg PG/g sample, respectively, which were mostly included in the organic solvent partitioned fractions. Strong NOI activity was observed in the EtOH extracts and their solvent partitioned fractions from D. viridis and C. filum. In addition, the EtOH extract and its solvent partitioned fractions of D. viridis exhibited little cytotoxicity to Raw 264.7 cells. The most potent ABTS and DPPH radical scavenging capacity was shown in the EtOH extracts and their solvent partitioned fractions from S. lomentaria and C. filum, and both also exhibited strong reducing ability. In the PCA analysis the content of TPT had a good correlation with DPPH ( r = 0.62), ABTS ( r = 0.69) and reducing power ( r = 0.65), however, an unfair correlation was observed between the contents of TP and TPT and NOI, suggesting that the phlorotannins might be responsible for the DPPH and ABTS radical scavenging activities.

Cho, Myoung Lae; Lee, Dong-Jin; Lee, Hyi-Seung; Lee, Yeon-Ju; You, Sang Guan

2013-12-01

146

Molecular characterization and expression analysis of a putative LPS-induced TNF-alpha factor (LITAF) from pearl oyster Pinctada fucata.  

PubMed

The lipopolysaccharide-induced TNF-alpha factor (LITAF) is a novel transcription factor, which plays an important role in regulating the expression of TNF-alpha and various inflammatory cytokines in response to LPS stimulation and forms a dependent signaling pathway separated from NF-kappaB. Herein, we described the identification and characterization of pearl oyster Pinctada fucata LPS-induced TNF-alpha factor gene (designated as poLITAF). The poLITAF cDNA was 932 bp long and consisted of a 5'-untranslated region (UTR) of 45 bp, a 3'-UTR of 497 bp with two cytokine RNA instability motifs (ATTTA), and an open reading frame (ORF) of 390 bp encoding a polypeptide of 129 amino acids with an estimated molecular mass of 13.5 kDa and a theoretical isoelectric point of 8.36. A LITAF domain at C-terminal was identified in the poLITAF using SMART analysis, which contained two conserved CXXC motifs. Homology analysis of the deduced amino acid sequence of the poLITAF with other known LITAF sequences by MatGAT software revealed that the poLITAF shared 44.2-67.4% similarity and 35.4-50.0% identity to the other known LITAF sequences. The expression level of poLITAF mRNA was the highest in digestive gland and gill, moderate in adductor muscle, gonad, intestine and mantle, the lowest in haemocytes. The poLITAF mRNA expression was significantly up-regulated at 24 h in gill and at 12 h in digestive gland after LPS stimulation respectively. These results suggested that the poLITAF was a constitutive and inducible acute-phase protein that perhaps involved in the innate immune response of pearl oyster. PMID:19426809

Zhang, Dianchang; Jiang, Jingjing; Jiang, Shigui; Ma, Jianjun; Su, Tianfeng; Qiu, Lihua; Zhu, Caiyan; Xu, Xinping

2009-09-01

147

PGE 2 exerts its effect on the LPS-induced release of TNF-?, ET1, ILl?, IL6 and IL10 via the EP2 and EP4 receptor in rat liver macrophages  

Microsoft Academic Search

Lipopolysaccharide (LPS) induces a release of tumor necrosis factor (TNF)-?, endothelin (ET)-1, interleukin (IL)-l?, IL-6 and IL-10 in rat liver macrophages (Kupffer cells). Prostaglandin (PG)E2 inhibits the release of the fibrogenic mediators TNF-?, ET-1 and IL-l?, and enhances the release of the anti-fibrogenic mediators IL-6 and IL-10. This effect of PGE2 is mimicked by specific agonists for the PGE2 receptors

Lars Treffkorn; Roland Scheibe; Takayuki Maruyama; Peter Dieter

2004-01-01

148

PARP-1 Inhibitor, DPQ, Attenuates LPS-Induced Acute Lung Injury through Inhibiting NF-?B-Mediated Inflammatory Response  

PubMed Central

Acute lung injury (ALI) is characterized by overwhelming lung inflammation and anti-inflammation treatment is proposed to be a therapeutic strategy for ALI. Poly (ADP-ribose) polymerase-1 has been demonstrated to be involved in tissue inflammation and one of its inhibitors, 3, 4-Dihydro-5[4-(1-piperindinyl)butoxy]-1(2H)-isoquinoline (DPQ), exerts anti-inflammatory effect. However, it is still unclear whether the DPQ possesses the protective effect on ALI and what mechanisms are involved. In this study, we tested the effect of DPQ on the lung inflammation induced by lipopolysaccharide (LPS) challenge in mice. We found that 6 h-LPS challenge induced significant lung inflammation and vascular leakage in mice. Treatment with DPQ at the dose of 10 ?g/kg markedly reduced the neutrophil infiltration, myeloperoxidase activity and up-regulation of pro-inflammatory mediators and cytokines. LPS-elevated vascular permeability was decreased by DPQ treatment, accompanied by the inhibition of apoptotic cell death in mice lungs. In addition, we isolated mice peritoneal macrophages and showed pretreatment with DPQ at 10 ?M inhibited the production of cytokines in the macrophages following LPS stimulation. DPQ treatment also inhibited the phosphorylation and degradation of I?B-?, subsequently blocked the activation of nuclear factor (NF)-?B induced by LPS in vivo and in vitro. Taken together, our results show that DPQ treatment inhibits NF-?B signaling in macrophages and protects mice against ALI induced by LPS, suggesting inhibition of Poly (ADP-ribose) polymerase-1 may be a potential and effective approach to resolve inflammation for the treatment of ALI. PMID:24278171

Wang, Gang; Huang, Xiaojia; Li, Yongjin; Guo, Kangkang; Ning, Pengbo; Zhang, Yanming

2013-01-01

149

A molecular pharmacology study into the anti-inflammatory actions of Euphorbia hirta L. on the LPS-induced RAW 264.7 cells through selective iNOS protein inhibition.  

PubMed

Euphorbia hirta L. has been widely used in India and Chinese society. The molecular pharmacology basis of its anti-inflammatory effect is revealed in this work. The ethanol extract of Euphorbia hirta L. (Eh) and its active component were studied in lipopolysaccharide (LPS)-activated macrophage cells (RAW 264.7) as an established inflammation model. After activation, nitric oxide (NO) production and expression of iNOS protein and iNOS mRNA were measured by using a colorimetric assay (Griess reagent), western blotting, and reverse transcription polymerase chain reaction (RT-PCR), respectively. The alteration in the content of PGE(2), TNFalpha, and IL-6 was concurrently monitored by ELISA. In results, we found that in the concentration range without showing cytotoxicity, Eh produced a remarkable anti-inflammatory effect via its active component of beta-amyrin and showed a dose-related inhibition of LPS-induced NO production. This phenomenon is in accordance with a substantial inhibition of iNOS protein. However, the expression of iNOS gene was unaffected by Eh treatments. Compared with indomethacin, Eh has much more potency and a specific action of NO inhibition but Eh works less specifically on PGE(2), IL-6, and TNF-alpha inhibition. The extract of Euphorbia hirta L. and its component beta-amyrin are able to block most of the iNOS protein functions and NO induction, and could therefore be new selective NO inhibitors with great potential in treating arthritis inflammation. PMID:20390370

Shih, Mei-Fen; Cheng, Yih-Dih; Shen, Chia-Rui; Cherng, Jong-Yuh

2010-07-01

150

Furanodien-6-one from Commiphora erythraea inhibits the NF-?B signalling and attenuates LPS-induced neuroinflammation.  

PubMed

We investigated the in vitro anti-inflammatory activity of 1(10),4-furanodien-6-one, one the most active compounds of the hexane extract of Commiphora erythraea (Ehrenb.) Engl., by exposing microglial BV-2 cells to lipopolysaccharide. We showed that furanodien-6-one pre-treatment restored cell viability and ROS to control levels while halving NO generation. Production of pro-inflammatory IL-6, IL-23, IL-17, TGF-?, and INF-?, significantly induced by LPS, was also markedly reduced by furanodien-6-one treatment. We further showed that furanodien-6-one protects primary neuronal cultures against the inflammatory/toxic insults of LPS-treated BV-2 conditioned media, indicating that furanodien-6-one exerts anti-inflammatory/cytoprotective effects in neuronal cells. We then investigated whether furanodien-6-one exerts anti-inflammatory properties in an in vivo model of microglial activation. In adult mice ip-injected with LPS we found that furanodien-6-one had strong cerebral anti-inflammatory properties by inhibiting liver and brain TNF? as well as IL-1? expression. Results were not unexpected since FTIR-metabolomic analyses showed that furanodien-6-one-treated mice had a reduced dissimilarity to control animals and that the response to LPS treatment was markedly modified by furanodien-6-one. In conclusion our data provide strong evidence of the anti-inflammatory properties of furanodien-6-one that could be exploited to counteract degenerative pathologies based on neuroinflammation. PMID:23357788

Bellezza, Ilaria; Mierla, Annalisa; Grottelli, Silvia; Marcotullio, Maria Carla; Messina, Federica; Roscini, Luca; Cardinali, Gianluigi; Curini, Massimo; Minelli, Alba

2013-07-01

151

Identification of a LPS-induced TNF-? factor (LITAF) from mollusk Solen grandis and its expression pattern towards PAMPs stimulation.  

PubMed

Lipopolysaccharide-induced TNF-? factor (LITAF) is one of the most important transcription factors mediating TNF-? transcription. In the present study, a LITAF gene (designated as SgLITAF) was identified from razor clams Solen grandis. The full-length cDNA of SgLITAF was of 1476 bp, encoding a polypeptide of 130 amino acids showed high similarity to other known LITAFs. SgLITAF encoded a LITAF domain and the Zn(2+)-binding motifs in the domain were well conserved. The mRNA transcripts of SgLITAF were detected in all tested tissues of healthy razor clams, including mantle, gill, gonad, hemocytes, muscle and hepatopancreas, and with the highest expression level in hepatopancreas. The expression level of SgLITAF in hemocytes was significantly up-regulated (P < 0.01) after razor clams were stimulated by LPS or ?-1, 3-glucan, but no obvious fluctuation of SgLITAF mRNA expression was observed after PGN stimulation. All the results indicated that there might be a LITAF-regulated TNF-? signaling pathway existing in S. grandis, which involved in the immune response not only against gram-negative bacteria but also towards fungi. PMID:23891855

Yang, Dinglong; Wei, Xiumei; Yang, Jianmin; Yang, Jialong; Xu, Jie; Fang, Jinghui; Wang, Sheng; Liu, Xiangquan

2013-10-01

152

Dexamethasone attenuates LPS-induced changes in expression of urea transporter and aquaporin proteins, ameliorating brain endotoxemia in mice  

PubMed Central

Aim: AQP4 in the brain is involved in the occurrence and development of a variety of encephalopathy. AQPs family changes in kidney were accompanied by altered UTs family. The aim of this study was to observe AQP4 and UT-A3 expression in CNS and to explore their role in the pathogenesis of endotoxemia encephalopathy following peripheral LPS injection in mice. Methods: Endotoxemia was induced in C57Bl/6 mice by intraperitoneal injection of LPS. The expression of UT-A3 and AQP4 in brain were detected by Western blot and immunohistochemistry, the level of cytokines were detected by ELISA, and the content of LDH, AST/ALT, BUN and CREA were detected by colorimetric method. Results: As compared with the control group, in model group, the brain weight/ body weight ratio increased by 13%. Meanwhile, a 2.5 fold increase in LDH and a 1.2 fold increase in AST/ALT were found in peripheral serum (P < 0.05), and also, BUN and CREA increased 2.5 fold (P < 0.01). In addition to severe CNS injury in response to lipopolysaccharide, the contents of cytokines and the expression of AQP4 protein in hippocampal is increased (P < 0.05), while the expression of UT-A3 protein in the hippocampus and cortical astrocytes decreased (P < 0.05). And, in part, Dexa pretreatment attenuated those effects. Conclusions: In endotoxemia encephalopathy, AQPs and UTs which regulate the functions of cell membrane are both altered. We suggested that the molecular mechanisms of regulation in endotoxemia may provide a new strategy for clinical treatment of the disease and drug binding sites. PMID:25674208

Du, Yanwei; Meng, Yan; Lv, Xuejiao; Guo, Lirong; Wang, Xiaoqin; Su, Zhenzhong; Li, Lu; Li, Na; Zhao, Shuhua; Zhao, Lijing; Zhao, Xuejian

2014-01-01

153

Inhibition of lipopolysaccharide-induced inducible nitric oxide synthase and cyclooxygenase-2 expression by xanthanolides isolated from Xanthium strumarium.  

PubMed

Three sesquiterpenoids, xanthatin (1), xanthinosin (2), and 4-oxo-bedfordia acid (3) were isolated from Xanthium strumarium as inhibitors of nitric oxide synthesis in activated microglia (IC(50) values: 0.47, 11.2, 136.5 microM, respectively). Compounds 1 and 2 suppressed the expression of iNOS and COX-2 and the activity of NF-kappaB through the inhibition of LPS-induced I-kappaB-alpha degradation in microglia. PMID:18276135

Yoon, Jeong Hoon; Lim, Hyo Jin; Lee, Hwa Jin; Kim, Hee-Doo; Jeon, Raok; Ryu, Jae-Ha

2008-03-15

154

Inhibition of lipopolysaccharide-induced inducible nitric oxide synthase and cyclooxygenase-2 expression by xanthanolides isolated from Xanthium strumarium  

Microsoft Academic Search

Three sesquiterpenoids, xanthatin (1), xanthinosin (2), and 4-oxo-bedfordia acid (3) were isolated from Xanthium strumarium as inhibitors of nitric oxide synthesis in activated microglia (IC50 values: 0.47, 11.2, 136.5?M, respectively). Compounds 1 and 2 suppressed the expression of iNOS and COX-2 and the activity of NF-?B through the inhibition of LPS-induced I-?B-? degradation in microglia.

Jeong Hoon Yoon; Hyo Jin Lim; Hwa Jin Lee; Hee-Doo Kim; Raok Jeon; Jae-Ha Ryu

2008-01-01

155

The sphingosine kinase 1 and S1P1 axis specifically counteracts LPS-induced IL-12p70 production in immune cells of the spleen.  

PubMed

Sphingosine-1-phosphate (S1P) has been implicated in angiogenesis, inflammation, cancerogenesis, neurological excitability and immune regulation and is synthesized by two different sphingosine kinases (SphK). It was suggested that mice lacking the gene for SphK1 exhibit no obvious phenotype, because SphK2 compensates for its absence. However, recent investigations revealed that under challenge SphK1 contributed to pro-inflammatory processes favoring Th2 and Th17 rather than Th1-type reactions. To investigate the immune modulatory role of SphK1 as opposed to SphK2 specifically for the Th1 propagating IL-12p70 we compared WT and SphK1(-/-) splenocytes and Flt3-ligand differentiated BMCs of WT and SphK1(-/-), representing dendritic cells as major producers of IL-12p70, incubated with LPS. We determined the impact on IL-12p70 in comparison to other inflammatory cytokines, and on DC and macrophage surface marker expression, SphK mRNA, protein expression and enzymatic activity in splenocytes. Our data demonstrated that SphK1 deficiency enhanced LPS-induced IL-12p70 production although SphK2 was present. To further characterize SphK1-dependent IL-12p70 regulation we exogenously applied S1P, SEW2871 and the new potent S1P1 agonist CYM5442. Both S1P and S1P1-specific analogs fully compensated the increase of IL-12p70 production in SphK1-deficient splenocytes. The use of pertussis toxin, to block G(i)-coupled signaling downstream of S1P1, again increased IL-12p70 and neglected the compensation achieved by addition of S1P and S1P1 agonists pointing on the importance of this specific S1P-receptor. Given that, in parallel to a prominent IL-12p35 increase following LPS stimulation, LPS also enhanced SphK expression and total SphK activity, we concluded that SphK1-derived S1P acting via S1P1 is a major mechanism of this negative IL-12p70 feedback loop, which did not affect other cytokines. Moreover, our data showed that SphK2 activity failed to compensate for SphK1 deficiency. These findings clearly point to a divergent and cytokine-specific impact of immune cell SphK1 and SphK2 in chronic inflammation and cancer. PMID:21435724

Schröder, Matthias; Richter, Cornelia; Juan, Martina Herrero San; Maltusch, Katrin; Giegold, Oliver; Quintini, Gianluca; Pfeilschifter, Josef M; Huwiler, Andrea; Radeke, Heinfried H

2011-05-01

156

Pseudoguaianolides isolated from Inula britannica var. chinenis as inhibitory constituents against inducible nitric oxide synthase.  

PubMed

Three pseudoguaianolide type sesquiterpenes, bigelovin (1), 2,3-dihydroaromaticin (2), and ergolide (3) were isolated as inhibitory constituents against inducible nitric oxide synthase (iNOS) from the flowers of Inula britannica var. chinensis. Bigelovin (1) exhibited a highly potent inhibitory activity on lipopolysaccharide (LPS)-induced iNOS in murine macrophage RAW 264.7 cells with an IC50 value of 0.46 mM, which is about 8 times more potent than the known selective inhibitor of iNOS, L-N6-(1-iminoethyl)lysine (IC50 3.49 microM). 2,3-Dihydroaromaticin (2) and ergolide (3) also exhibited potent inhibitory activities on LPS-induced iNOS with IC50 values of 1.05 and 0.69 microM, respectively. PMID:12009027

Lee, Hyun-Tai; Yang, Seung-Won; Kim, Kyeong Ho; Seo, Eun-Kyoung; Mar, Woongchon

2002-04-01

157

Synthesis and effects of new caffeic acid derivatives on nitric oxide production in lipopolysaccharide-induced RAW 264.7 macrophages  

PubMed Central

In this study, 20 new derivatives of caffeic acid esters were synthesized and their inhibitory activities against the lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW264.7 macrophages were determined. Compounds 3l, 3r, 3s and 3t were found to decrease nitrite levels in a dose-dependent manner in LPS-induced cells and showed potent inhibitory activities against the NO production in RAW264.7 macrophages with IC50 values of 7.4, 5.9, 3.3 and 2.2 ?M, respectively. They could be selected as compromising compounds for the later pharmacological study. PMID:24955176

Zhang, Jie; Xu, Liu-Xin; Xu, Xu-Sheng; Li, Bo-Wei; Wang, Rui; Fu, Jian-Jun

2014-01-01

158

Gallotannin isolated from Euphorbia species, 1,2,6-tri-O-galloyl-beta-D-allose, decreases nitric oxide production through inhibition of nuclear factor-kappa>B and downstream inducible nitric oxide synthase expression in macrophages.  

PubMed

Gallotannins are plant secondary metabolites and are widely included to polyphenolic compounds. Gallotannins are water-soluble polyphenols with wide-ranging biological activities. Nitric oxide (NO) is well known as a mediator of inflammation. Macrophages express inducible nitric oxide synthase (iNOS) and produce NO after lipopoly saccharide (LPS) stimulation. In the present study, we examined the inhibitory effects of seven gallotannins isolated from Euphorbia species (Euphorbiaceae) on the LPS-induced NO production and underlying mechanisms of action. Among the seven gallotannins, 1,2,3,4,6-penta-O-galloyl-beta-D-glucose (gallotannin 15) and 1,2,6-tri-O-galloyl-beta-D-allose (gallotannin 23) significantly reduced LPS-induced NO production in macrophages. Gallotannin 15 and 23 (0.1-10 microg/ml) dose-dependently decreased gene expression and production of iNOS. In addition, gallotannin 15 and 23 (0.1-10 microg/ml) dose-dependently inhibited LPS-induced activation of nuclear factor (NF)-kappaB as indicated by inhibition of degradation of I-kappaBalpha, nuclear translocation of NF-kappaB, and NF-kappaB-dependent gene reporter assay. Our results suggest that gallotannins possess an inhibitory effect on the LPS-induced inflammatory reaction. PMID:19483314

Kim, Mi-Sun; Park, Seung-Bin; Suk, Kyoungho; Kim, In Kyeom; Kim, Sang-Yong; Kim, Jeong-Ah; Lee, Seung Ho; Kim, Sang-Hyun

2009-06-01

159

Ethanol extract of Synurus deltoides (Aiton) Nakai suppresses in vitro LPS-induced cytokine production in RAW 264.7 macrophages and in vivo acute inflammatory symptoms.  

PubMed

Synurus deltoides (Aiton) Nakai, belonging to the Compositae family, is an edible plant widely distributed in Northeast Asia. In this study, we examined the mechanisms underlying the immunomodulative effects of the ethanol extract of S. deltoides (SDE). The SDE extract strongly down-regulated the mRNA expression of the inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and tumour necrosis factor (TNF)-?, thereby inhibiting the production of nitric oxide (NO), prostaglandin E2 (PGE2), and TNF-? in the lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Furthermore, SDE also suppressed the nuclear translocation of the activation protein (AP)-1 and the nuclear factor-?B (NF-?B), and simultaneously decreased the phosphorylation of extracellular signal-regulated protein kinases (ERK), p38, and Akt. In agreement with the in vitro observations, the orally administered SDE ameliorated the acute inflammatory symptoms in the arachidonic acid-induced ear edema and the EtOH/HCl-induced gastritis in mice. Therefore, S. deltoides have a potential anti-inflammatory capacity in vitro and in vivo, suggesting the potential therapeutic use in the inflammation-associated disorders. PMID:24611100

Jiang, Yunyao; Wang, Myeong-Hyeon

2014-02-01

160

Short-term heating reduces the anti-inflammatory effects of fresh raw garlic extracts on the LPS-induced production of NO and pro-inflammatory cytokines by downregulating allicin activity in RAW 264.7 macrophages.  

PubMed

Garlic has a variety of biologic activities, including anti-inflammatory properties. Although garlic has several biologic activities, some people dislike eating fresh raw garlic because of its strong taste and smell. Therefore, garlic formulations involving heating procedures have been developed. In this study, we investigated whether short-term heating affects the anti-inflammatory properties of garlic. Fresh and heated raw garlic extracts (FRGE and HRGE) were prepared with incubation at 25 °C and 95 °C, respectively, for 2 h. Treatment with FRGE and HRGE significantly reduced the LPS-induced increase in the pro-inflammatory cytokine concentration (TNF-?, IL-1?, and IL-6) and NO through HO-1 upregulation in RAW 264.7 macrophages. The anti-inflammatory effect was greater in FRGE than in HRGE. The allicin concentration was higher in FRGE than in HRGE. Allicin treatment showed reduced production of pro-inflammatory cytokines and NO and increased HO-1 activity. The results show that the decrease in LPS-induced NO and pro-inflammatory cytokines in RAW 264.7 macrophages through HO-1 induction was greater for FRGE compared with HRGE. Additionally, the results indicate that allicin is responsible for the anti-inflammatory effect of FRGE. Our results suggest a potential therapeutic use of allicin in the treatment of chronic inflammatory disease. PMID:23583806

Shin, Jung-Hye; Ryu, Ji Hyeon; Kang, Min Jung; Hwang, Cho Rong; Han, Jaehee; Kang, Dawon

2013-08-01

161

Inhibition of LPS-induced TNF-? and NO production in mouse macrophage and inflammatory response in rat animal models by a novel Ayurvedic formulation, BV-9238.  

PubMed

Rheumatoid arthritis is a chronic crippling disease, where protein-based tumor necrosis factor-alpha (TNF-?) inhibitors show significant relief, but with potentially fatal side effects. A need for a safe, oral, cost-effective small molecule or phyto-pharmaceutical is warranted. BV-9238 is an Ayurvedic poly-herbal formulation containing specialized standardized extracts of Withania somnifera, Boswellia serrata, Zingiber officinale and Curcuma longa. The anti-inflammatory and anti-arthritic effects of BV-9238 were evaluated for inhibition of TNF-? and nitric oxide (NO) production, in lipopolysaccharide-stimulated, RAW 264.7, mouse macrophage cell line. BV-9238 reduced TNF-? and NO production, without any cytotoxic effects. Subsequently, the formulation was tested in adjuvant-induced arthritis (AIA) and carrageenan-induced paw edema (CPE) rat animal models. AIA was induced in rats by injecting Freund's complete adjuvant intra-dermally in the paw, and BV-9238 and controls were administered orally for 21?days. Arthritic scores in AIA study and inflamed paw volume in CPE study were significantly reduced upon treatment with BV-9238. These results suggest that the anti-inflammatory and anti-arthritic effects of BV-9238 are due to its inhibition of TNF-?, and NO, and this formulation shows promise as an alternate therapy for inflammatory disorders where TNF-? and NO play important roles. PMID:24706581

Dey, Debendranath; Chaskar, Sunetra; Athavale, Nitin; Chitre, Deepa

2014-10-01

162

Extracts of Ficus exasperata leaf inhibit topical and systemic inflammation in rodents and suppress LPS-induced expression of mediators of inflammation in macrophages.  

PubMed

The leaves of Ficus exasperata are mashed and prepared as poultices that are placed on swellings, wounds, and arthritic joints to relieve swelling and pains by the Igede tribal community of Nigeria. The leaf and stalk are also squeezed and used to mitigate itching or inflammation. These claimed benefits inspired this study in which topical and systemic (acute, chronic) anti-inflammatory activities of a methanol/methylene chloride leaf extract of F. exasperata (MFE) were assessed in rodents. Effects of an aqueous leaf extract (AFE) on lipopolysaccharide-induced expression of interleukin-1? (IL-1?), tumor necrosis factor (TNF)-?, and inducible nitric oxide (iNO) were also investigated in murine bone marrow-derived macrophage (BMDM) cultures. Treatment of rats with MFE (200 and 400?mg/kg) led to significant inhibition of acute and chronic inflammation induced by, respectively, agar and formaldehyde in the paws. Topically, pre-application of mice with MFE (5 µg/ear) also significantly inhibited (by up to 21%) ear edema induced by xylene. In vitro, pre-treatment of BMDM with 5-100 µg AFE/ml significantly inhibited IL-1?, TNF?, and iNO production in a dose-related manner. BMDM viability was not significantly affected AFE at concentrations up to 200 µg/ml. Initial studies showed that flavonoids, alkaloids, and terpenoids were the predominant phytoconstituents in each extract. In conclusion, the results of the various investigations indicated that F. exasperata leaf extracts possess anti-inflammatory properties that could underlie the benefits associated with the folklore use of the plant. The results also show that the extracts may be acting through a suppression of mediators of inflammation, such as IL-1?, TNF?, and iNO. PMID:23098056

Nworu, Chukwuemeka S; Nwuke, Henry C; Akah, Peter A; Okoye, Festus B C; Esimone, Charles O

2013-01-01

163

Inhibition of LPS-induced TLR4 signaling products in murine macrophages by phenylmethimazole: an assay methodology for screening potential phenylmethimazole analogs.  

PubMed

Preclinical Research Phenylmethimazole (C10) is an inhibitor of Toll-like receptor (TLR3 and TLR4) expression and signaling. In this study, we carried out a detailed investigation of the effect of C10 on TLR4 and its molecular signaling products in RAW 264.7 macrophages using quantitative real-time polymerase chain reaction (PCR), ELISA and cell toxicity assays, a set of in vitro assays that may be used to screen future C10 analogs. C10 exhibited an inhibitory effect on TLR4 MyD88-dependent and MyD88-independent pathways. Within the TLR4 pathway, C10 inhibited the expression of cytokines, cytokine receptors, kinases, adapter molecules and transcription factors, suggesting a pathway-wide inhibitory effect. We also found that C10 dose-dependently inhibited the expression of TLR4 signaling products, specifically IL-6, inducible nitric oxide (NO) synthase and IFN?. Additionally, pre-treatment of RAW 264.7 cells with C10 resulted in protection from lipopolysaccharide (LPS) insults, suggesting C10 may be bound to the target thus exhibiting activity during/following LPS stimulation. Also, dimethyl sulfoxide, the solvent for C10 exhibited inhibitory effect on TLR4 signaling products independent from the effects of C10. Combined, this study enhances understanding of the actions of action on TLR4 signaling pathway providing a path for the development of new C10 analogs for inhibiting TLR expression and signaling. PMID:25408546

Deosarkar, Sudhir P; Bhatt, Pooja; Gillespie, Jessica; Goetz, Douglas J; McCall, Kelly D

2014-12-01

164

Inhibitory effects of natural sesquiterpenoids isolated from the rhizomes of Curcuma zedoaria on prostaglandin E2 and nitric oxide production.  

PubMed

Beta-turmerone and ar-turmerone, sesquiterpenoids isolated from the rhizome of Curcuma zedoaria, inhibited lipopolysaccharide (LPS)-induced prostaglandin E 2 production in cultured mouse macrophage cell RAW 264.7 in a dose-dependent manner (IC 50 = 7.3 microM for beta-turmerone; IC 50 = 24.0 microM for ar-turmerone). In addition, these compounds exhibited inhibitory effects on LPS-induced nitric oxide production in the cell system. PMID:12094302

Hong, Chae Hee; Noh, Min Soo; Lee, Woo Young; Lee, Sang Kook

2002-06-01

165

Pepsin-pancreatin protein hydrolysates from extruded amaranth inhibit markers of atherosclerosis in LPS-induced THP-1 macrophages-like human cells by reducing expression of proteins in LOX-1 signaling pathway  

PubMed Central

Background Atherosclerosis is considered a progressive disease that affects arteries that bring blood to the heart, to the brain and to the lower end. It derives from endothelial dysfunction and inflammation, which play an important role in the thrombotic complications of atherosclerosis. Cardiovascular disease is the leading cause of death around the world and one factor that can contribute to its progression and prevention is diet. Our previous study found that amaranth hydrolysates inhibited LPS-induced inflammation in human and mouse macrophages by preventing activation of NF-?B signaling. Furthermore, extrusion improved the anti-inflammatory effect of amaranth protein hydrolysates in both cell lines, probably attributed to the production of bioactive peptides during processing. Therefore, the objective of this study was to compare the anti-atherosclerotic potential of pepsin-pancreatin hydrolysates from unprocessed and extruded amaranth in THP-1 lipopolysaccharide-induced human macrophages and suggest the mechanism of action. Results Unprocessed amaranth hydrolysate (UAH) and extruded amaranth hydrolysate (EAH) showed a significant reduction in the expression of interleukin-4 (IL-4) (69% and 100%, respectively), interleukin-6 (IL-6) (64% and 52%, respectively), interleukin-22 (IL-22) (55% and 70%, respectively). Likewise, UAH and EAH showed a reduction in the expression of monocyte-chemo attractant protein-1 (MCP-1) (35% and 42%, respectively), transferrin receptor-1 (TfR-1) (48% and 61%, respectively), granulocyte-macrophage colony-stimulating factor (GM-CSF) (59% and 63%, respectively), and tumor necrosis factor-? (TNF-?) (60% and 63%, respectively). Also, EAH reduced the expression of lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) (27%), intracellular adhesion molecule-1 (ICAM-1) (28%) and matrix metalloproteinase-9 (MMP-9) (19%), important molecular markers in the atherosclerosis pathway. EAH, led to a reduction of 58, 52 and 79% for LOX-1, ICAM-1 and MMP-9, respectively, by confocal microscopy. Conclusions Extruded amaranth hydrolysate showed potential anti-atherosclerotic effect in LPS-induced THP-1 human macrophage-like cells by reducing the expression of proteins associated with LOX-1 signaling pathway. PMID:24891839

2014-01-01

166

SGLT-1-mediated glucose uptake protects intestinal epithelial cells against LPS-induced apoptosis and barrier defects: a novel cellular rescue mechanism?  

Microsoft Academic Search

Excessive apoptosis induced by enteric microbes leads to epithelial barrier defects. This mech- anism has been implicated in the pathogenesis of inflammatory bowel diseases (IBD) and bacterial enter- itis. The sodium-dependent glucose cotransporter (SGLT-1) is responsible for active glucose uptake in enterocytes. The aim was to investigate the effects of SGLT-1 glucose uptake on enterocyte apoptosis and barrier defects induced

Linda C. H. Yu; Andrew N. Flynn; Jerrold R. Turner; Andre G. Buret

2005-01-01

167

Astrocytic TLR4 expression and LPS-induced nuclear translocation of STAT3 in the sensory circumventricular organs of adult mouse brain.  

PubMed

The sensory circumventricular organs (CVOs) comprise the organum vasculosum of the lamina terminalis (OVLT), subfornical organ (SFO), and area postrema (AP) and lack the blood-brain barrier. The expression of Toll-like receptor 4 (TLR4) was seen at astrocytes throughout the sensory CVOs and at microglia in the AP and solitary nucleus around the central canal. The peripheral and central administration of lipopolysaccharide induced a similar pattern of nuclear translocation of STAT3. A microglia inhibitor minocycline largely suppressed lipopolysaccharide-induced astrocytic nuclear translocation of STAT3 in the OVLT and AP, but its effect was less in the SFO. PMID:25595264

Nakano, Yousuke; Furube, Eriko; Morita, Shoko; Wanaka, Akio; Nakashima, Toshihiro; Miyata, Seiji

2015-01-15

168

Plasminogen activator inhibitor type-1 deficiency exaggerates LPS-induced acute lung injury through enhancing Toll-like receptor 4 signaling pathway.  

PubMed

Mice lacking plasminogen activator inhibitor-1 (PAI-1) did not affect lung injury induced by gram-positive bacteria pneumococcal pneumonia but worsened lung injury induced by gram-negative bacteria Klebsiella. The exact mechanisms have not been completely elucidated. In this study, we examined the signaling pathway of Toll-like receptor 4 (TLR4) with/without PAI-1 in acute lung injury (ALI) induced by lipopolysaccharides (LPS) in mice. PAI-1 knockout mice (n=60) and wild-type mice (n=60) were exposed to LPS intratracheal instillation. Different groups of mice were then sacrificed at 0 and 8 h after LPS instillation. PAI-1-/- mice showed increased excess lung water and elevated cytokines production and release. In addition, expression of TLR4 was up-regulated and the phosphorylation activation of extracellular regulating kinase (ERK) and c-Jun N-terminal kinase (JNK) were also increased in PAI-1 knockout mice compared to wild-type mice. Inversely, interleukin (IL)-1 receptor-associated kinase-M (IRAK-M) and suppressor of cytokine signaling 1 (SOCS1) were both significantly reduced in PAI-1-/-mice after LPS challenge. PAI-1 deletion increased lung injury induced by LPS through up-regulation of TLR4, ERK and C-JNK and down-regulation of TLR4 negative regulators. PMID:21577093

Hua, Feng; Ren, Weiying; Zhu, Lei

2011-09-01

169

LPS-induced Expression of MMPs in Human Monocytes is Suppressed by IFN-? via Superinduction of ATF-3 and Suppression of AP-1 Proteins  

PubMed Central

Matrix metalloproteinases (MMPs) are induced during inflammatory responses and are important for immune regulation, angiogenesis, wound healing and tissue remodeling. Expression of MMPs needs to be tightly controlled to avoid excessive tissue damage. In this study we investigated the regulation of MMP expression by inflammatory factors in primary human monocytes and macrophages. IFN?, which augments inflammatory cytokine production in response to macrophage-activating factors such as Toll-like receptor (TLR) ligands, instead broadly suppressed TLR-induced MMP expression. Inhibition of MMP expression was dependent on STAT1 and required de novo protein synthesis. IFN? strongly enhanced TLR-induced expression of the transcriptional repressor ATF-3 in a STAT1-dependent manner, which correlated with recruitment of ATF-3 to the endogenous MMP-1 promoter as detected by chromatin immunoprecipitation assays. RNA interference experiments further supported a role for ATF-3 in suppression of MMP-1 expression. In addition, IFN? suppressed DNA binding by AP-1 transcription factors that are known to promote MMP expression and a combination of supershift, RNA interference and overexpression experiments implicated AP-1 family member Fra-1 in the regulation of MMP-1 expression. These results define an IFN?-mediated homeostatic loop that limits the potential for tissue damage associated with inflammation, and identify transcriptional factors that regulate MMP expression in myeloid cells in inflammatory settings. PMID:18802113

Ho, Hao H.; Antoniv, Taras; Ji, Jong-Dae; Ivashkiv, Lionel B.

2008-01-01

170

Effects of phenylethanoid glycosides from Digitalis purpurea L. on the expression of inducible nitric oxide synthase.  

PubMed

We have isolated four different phenylethanoid glycosides (purpureaside A, desrhamnosyl acteoside, calceolarioside B and plantainoside D) from the leaves of Digitalis purpurea (foxglove). The effects of these glycosides on activator protein-1 (AP-1)-mediated inducible nitric oxide synthase (iNOS) gene expression in the Raw264.7 macrophage cell line have been studied. Of these four glycosides, purpureaside A potently inhibited iNOS induction by lipopolysaccharide (LPS). Increase in iNOS mRNA by LPS was completely suppressed by purpureaside A. Purpureaside A did not significantly affect LPS-inducible nuclear factor-kappaB (NF-kappaB) activation or the nuclear translocation of p65. Moreover, a reporter gene assay using AP-1 specific luciferase reporter revealed that the enhanced activity of AP-1 by LPS was completely abolished in cells treated with purpureaside A. These results demonstrated that purpureaside A inhibited LPS-inducible iNOS expression in macrophages through the suppression of AP-1, but not of NF-kappaB. PMID:15969951

Oh, Jae Wook; Lee, Jeong Yong; Han, Song Hee; Moon, Young Hee; Kim, Yoon Gyoon; Woo, Eun-Rhan; Kang, Keon Wook

2005-07-01

171

New phenylpropanoid and other compounds from Illicium lanceolatum with inhibitory activities against LPS-induced NO production in RAW 264.7 macrophages.  

PubMed

Illicium lanceolatum is a traditional Chinese medicine (TCM) for treating inflammatory diseases. Anti-inflammatory activities of I. lanceolatum stems and leaves were tested using ear edema models induced by dimethyl benzene in mice. Bioassay-guided fractionation of the ethanol extract of I. lanceolatum leaves and stems revealed that the ethyl acetate fraction exhibited inhibitory potency to dimethyl benzene-induced edema in the mouse ear. Phytochemical investigation on the active fraction led to the isolation of a new phenylpropanoid (1), together with fifteen known compounds. This is the first report of the isolation of 2-16 from I. lanceolatum. Of these compounds, compounds 1, 2 and 3 showed inhibitory activity on LPS-stimulated NO production in RAW 264.7 macrophages with IC50 values of 27.58, 26.59 and 34.35 ?g/mL, respectively. I. lanceolatum stems and leaves can be exploited to alleviate inflammatory diseases, which makes the rare medicinal plant resources sustainable. PMID:24613803

Gui, Xuan; Wang, Guowei; Zhang, Naidan; Huang, Baokang

2014-06-01

172

MR imaging and targeting of a specific alveolar macrophage subpopulation in LPS-induced COPD animal model using antibody-conjugated magnetic nanoparticles  

PubMed Central

Purpose Targeting and noninvasive imaging of a specific alveolar macrophage subpopulation in the lung has revealed the importance for early and better diagnosis and therapy of chronic obstructive pulmonary disease (COPD). In this study, the in vivo effect of pulmonary administration of iron oxide nanoparticles on the polarization profile of macrophages was assessed, and a noninvasive free-breathing magnetic resonance imaging (MRI) protocol coupled with the use of biocompatible antibody-conjugated superparamagnetic iron oxide (SPIO) nanoparticles was developed to enable specific targeting and imaging of a particular macrophage subpopulation in lipopolysaccharide-induced COPD mice model. Materials and methods Enzyme-linked immunosorbent assay, Real-time polymerase chain reaction, and flow cytometry analysis were performed to assess the biocompatibility of PEGylated dextran-coated SPIO nanoparticles. Specific biomarkers for M1 and M2 macrophages subsets were selected for conjugation with magnetic nanoparticles. MRI protocol using ultra-short echo time sequence was optimized to enable simultaneous detection of inflammation progress in the lung and detection of macrophages subsets. Flow cytometry and immunohistochemistry analysis were finally performed to confirm MRI readouts and to characterize the polarization profile of targeted macrophages. Results The tested SPIO nanoparticles, under the current experimental conditions, were found to be biocompatible for lung administration in preclinical settings. Cluster of differentiation (CD)86- and CD206-conjugated magnetic nanoparticles enabled successful noninvasive detection of M1 and M2 macrophage subpopulations, respectively, and were found to co-localize with inflammatory regions induced by lipopolysaccharide challenge. No variation in the polarization profile of targeted macrophages was observed, even though a continuum switch in their polarization might occur. However, further confirmatory studies are required to conclusively establish this observation. Conclusion Coupling of magnetic iron oxide nanoparticles with a specific antibody targeted to a particular macrophage subpopulation could offer a promising strategy for an early and better diagnosis of pulmonary inflammatory diseases using noninvasive MRI. PMID:24711699

Al Faraj, Achraf; Shaik, Asma Sultana; Afzal, Sibtain; Al Sayed, Baraa; Halwani, Rabih

2014-01-01

173

Suppression of LPS-Induced TNF-? Production in Macrophages by cAMP Is Mediated by PKA-AKAP95-p105  

PubMed Central

The activation of macrophages through Toll-like receptor (TLR) pathways leads to the production of a broad array of cytokines and mediators that coordinate the immune response. The inflammatory potential of this response can be reduced by compounds, such as prostaglandin E2, that induce the production of cyclic adenosine monophosphate (cAMP). Through experiments with cAMP analogs and multigene RNA interference (RNAi), we showed that key anti-inflammatory effects of cAMP were mediated specifically by cAMP-dependent protein kinase (PKA). Selective inhibitors of PKA anchoring, time-lapse microscopy, and RNAi screening suggested that differential mechanisms of PKA action existed. We showed a specific role for A kinase-anchoring protein 95 in suppressing the expression of the gene encoding tumor necrosis factor-?, which involved phosphorylation of p105 (also known as Nfkb1) by PKA at a site adjacent to the region targeted by inhibitor of nuclear factor ?B kinases. These data suggest that crosstalk between the TLR4 and cAMP pathways in macrophages can be coordinated through PKA-dependent scaffolds that localize specific pools of the kinase to distinct substrates. PMID:19531803

Wall, Estelle A.; Zavzavadjian, Joelle R.; Chang, Mi Sook; Randhawa, Baljinder; Zhu, Xiaocui; Hsueh, Robert C.; Liu, Jamie; Driver, Adrienne; Bao, Xiaoyan Robert; Sternweis, Paul C.; Simon, Melvin I.; Fraser, Iain D. C.

2009-01-01

174

Melatonin modulates microsomal PGE synthase 1 and NF-E2-related factor-2-regulated antioxidant enzyme expression in LPS-induced murine peritoneal macrophages  

PubMed Central

BACKGROUND AND PURPOSE Increasing evidence demonstrates that melatonin regulates inflammatory and immune processes acting as both an activator and inhibitor of these responses. Nevertheless, the molecular mechanisms of its anti-inflammatory action remain unclear. Here we have characterized the cellular mechanisms underlying the redox modulation of LPS-stimulated inflammatory responses in murine peritoneal macrophages by melatonin to provide insight into its anti-inflammatory effects. EXPERIMENTAL APPROACH Murine peritoneal macrophages were isolated and treated with melatonin in the presence or absence of LPS (5 ?g·mL?1) for 18 h. Cell viability was determined using sulforhodamine B assay and NO production was measured using the Griess reaction. Pro-inflammatory enzymes and transcription factors were detected by Western blotting. KEY RESULTS Without affecting cell viability, melatonin (12.5, 25, 50 and 100 ?M) reduced the level of nitrites, inducible NOS (iNOS), COX-2 and microsomal PGE synthase-1 (mPGES1) protein, and p38 MAPK phosphorylation, and prevented NF-?B translocation. Furthermore, melatonin treatment significantly increased NF-E2-related factor 2 (Nrf2) and haem oxygenase 1 (HO1) protein levels in murine macrophages exposed to LPS. CONCLUSIONS AND IMPLICATIONS Melatonin reduced pro-inflammatory mediators and enhanced the expression of HO1 via NF-?B, p38 MAPK and Nrf2 cascade signalling pathways in murine macrophages. Thus, melatonin might be a promising target for diseases associated with overactivation of macrophages. PMID:24116971

Aparicio-Soto, M; Alarcón-de-la-Lastra, C; Cárdeno, A; Sánchez-Fidalgo, S; Sanchez-Hidalgo, M

2014-01-01

175

Fluoxetine up-regulates expression of cellular FLICE-inhibitory protein and inhibits LPS-induced apoptosis in hippocampus-derived neural stem cell  

SciTech Connect

Fluoxetine is a widely used antidepressant compound which inhibits the reuptake of serotonin in the central nervous system. Recent studies have shown that fluoxetine can promote neurogenesis and improve the survival rate of neurons. However, whether fluoxetine modulates the proliferation or neuroprotection effects of neural stem cells (NSCs) needs to be elucidated. In this study, we demonstrated that 20 {mu}M fluoxetine can increase the cell proliferation of NSCs derived from the hippocampus of adult rats by MTT test. The up-regulated expression of Bcl-2, Bcl-xL and the cellular FLICE-inhibitory protein (c-FLIP) in fluoxetine-treated NSCs was detected by real-time RT-PCR. Our results further showed that fluoxetine protects the lipopolysaccharide-induced apoptosis in NSCs, in part, by activating the expression of c-FLIP. Moreover, c-FLIP induction by fluoxetine requires the activation of the c-FLIP promoter region spanning nucleotides -414 to -133, including CREB and SP1 sites. This effect appeared to involve the phosphatidylinositol-3-kinase-dependent pathway. Furthermore, fluoxetine treatment significantly inhibited the induction of proinflammatory factor IL-1{beta}, IL-6, and TNF-{alpha} in the culture medium of LPS-treated NSCs (p < 0.01). The results of high performance liquid chromatography coupled to electrochemical detection further confirmed that fluoxentine increased the functional production of serotonin in NSCs. Together, these data demonstrate the specific activation of c-FLIP by fluoxetine and indicate the novel role of fluoxetine for neuroprotection in the treatment of depression.

Chiou, S.-H. [Department of Medical Research and Education, Taipei Veterans General Hospital and National Yang-Ming University, Taipei, Taiwan (China) and Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan (China)]. E-mail: shchiou@vghtpe.gov.tw; Chen, S.-J. [Department of Ophthalmology, Taipei Veterans General Hospital and National Yang-Ming University, Taipei, Taiwan (China)]. E-mail: sjchen@vghtpe.gov.tw; Peng, C-H. [Institute of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan (China); Department of Ophthalmology, Shin Kong Wu Ho-Su Memorial Hospital, Taipei, Taiwan (China); Chang, Y.-L. [Department of Pharmacy, Taipei Veterans General Hospital and National Yang-Ming University, Taipei, Taiwan (China); Ku, H.-H. [Institute of Anatomy and Cell Biology, National Yang-Ming University, Taipei, Taiwan (China); Hsu, W.-M. [Department of Ophthalmology, Taipei Veterans General Hospital and National Yang-Ming University, Taipei, Taiwan (China); Ho, Larry L.-T. [Department of Medical Research and Education, Taipei Veterans General Hospital and National Yang-Ming University, Taipei, Taiwan (China); Lee, C.-H. [Department of Medical Research and Education, Taipei Veterans General Hospital and National Yang-Ming University, Taipei, Taiwan (China)

2006-05-05

176

Comparative analysis of the human hepatic and adipose tissue transcriptomes during LPS-induced inflammation leads to the identification of differential biological pathways and candidate biomarkers  

PubMed Central

Background Insulin resistance (IR) is accompanied by chronic low grade systemic inflammation, obesity, and deregulation of total body energy homeostasis. We induced inflammation in adipose and liver tissues in vitro in order to mimic inflammation in vivo with the aim to identify tissue-specific processes implicated in IR and to find biomarkers indicative for tissue-specific IR. Methods Human adipose and liver tissues were cultured in the absence or presence of LPS and DNA Microarray Technology was applied for their transcriptome analysis. Gene Ontology (GO), gene functional analysis, and prediction of genes encoding for secretome were performed using publicly available bioinformatics tools (DAVID, STRING, SecretomeP). The transcriptome data were validated by proteomics analysis of the inflamed adipose tissue secretome. Results LPS treatment significantly affected 667 and 483 genes in adipose and liver tissues respectively. The GO analysis revealed that during inflammation adipose tissue, compared to liver tissue, had more significantly upregulated genes, GO terms, and functional clusters related to inflammation and angiogenesis. The secretome prediction led to identification of 399 and 236 genes in adipose and liver tissue respectively. The secretomes of both tissues shared 66 genes and the remaining genes were the differential candidate biomarkers indicative for inflamed adipose or liver tissue. The transcriptome data of the inflamed adipose tissue secretome showed excellent correlation with the proteomics data. Conclusions The higher number of altered proinflammatory genes, GO processes, and genes encoding for secretome during inflammation in adipose tissue compared to liver tissue, suggests that adipose tissue is the major organ contributing to the development of systemic inflammation observed in IR. The identified tissue-specific functional clusters and biomarkers might be used in a strategy for the development of tissue-targeted treatment of insulin resistance in patients. PMID:21978410

2011-01-01

177

Characterization of two regulators of the TNF-? signaling pathway in Apostichopus japonicus: LPS-induced TNF-? factor and baculoviral inhibitor of apoptosis repeat-containing 2.  

PubMed

The TNF-? signaling cascade is involved in the regulation of a variety of biological processes, including cell proliferation, differentiation, apoptosis and the immune response in vertebrates. Here, two regulatory genes, lipopolysaccharide-induced tumor necrosis factor ? factor (LITAF) and baculoviral inhibitor of apoptosis repeat-containing 2 (BIRC2), were identified in coelomocytes from the sea cucumber Apostichopus japonicus by RNA-seq and RACE (denoted as AjLITAF and AjBIRC2, respectively). The full-length cDNA of AjLITAF was 1417?bp, with a 5' untranslated region (UTR) of 189?bp, a 3' UTR of 637?bp with one cytokine RNA instability motif (ATTTA) and an open reading frame (ORF) of 591?bp encoding a polypeptide of 196 amino acid residues and a predicted molecular weight of 22.1?kDa. The partial AjBIRC2 cDNA was 2324?bp with a 5' UTR of 145?bp, a 3' UTR of 469?bp and a complete ORF of 1710?bp encoding a polypeptide of 569 amino acid residues. Analysis of the deduced amino acid sequences revealed that both genes shared a remarkably high degree of structural conservation with their mammalian orthologs, including a highly conserved LITAF domain in AjLITAF and three types of BIR domains in AjBIRC2. Spatial expression analysis revealed that AjLITAF and AjBIRC2 were expressed at a slightly lower level in the intestine and tentacle tissues compared with the other four tissues examined. After challenging the sea cucumbers with Vibrio splendidus, the expression levels of AjLITAF and AjBIRC2 in coelomocytes were increased by 2.65-fold at 6?h and 1.76-fold at 24?h compared with the control group. In primary cultured coelomocytes, a significant increase in the expression of AjLITAF and AjBIRC2 was detected after 6?h of exposure to 1?µg mL(-1) LPS. Together, these results suggest that AjLITAF and AjBIRC2 might be involved in the sea cucumber immune response during the course of a pathogenic infection or exposure to pathogen-associated molecular pattern (PAMP) molecules. PMID:25307203

Zhang, Xiumei; Zhang, Pengjuan; Li, Chenghua; Li, Ye; Jin, Chunhua; Zhang, Weiwei

2015-01-01

178

SRC-FAMILY TYROSINE KINASE INHIBITOR (PP1) SUPPRESSES LPS-INDUCED EXPRESSION OF INDUCIBLE NITRIC OXIDE SYNTHASE MEDIATED THROUGH THE INHIBITION OF INTERFERON B EXPRESSION IN MACROPHAGES  

Technology Transfer Automated Retrieval System (TEKTRAN)

Bacterial lipopolysaccharide (LPS) activates Toll-like recptor (TLR4) leading to expression of inflammatory gene products. Non-receptor type Src-family tyrosine kinases are activated by LPS and associated with CD14, a co-receptor for LPS in monocytes and macrophages. Therefore, we determined the ro...

179

Bifunctional effects of fucoidan on the expression of inducible nitric oxide synthase  

SciTech Connect

Algal fucoidan is a marine sulfated polysaccharide with a wide variety of biological activities including anti-thrombotic and anti-inflammatory effects. This study evaluated the effect of fucoidan on the expression of inducible nitric oxide synthase (iNOS) in a macrophage cell line, RAW264.7. Low concentration range of fucoidan (10 {mu}g/ml) increased the basal expression level of iNOS in quiescent macrophages. However, we found for the first time that fucoidan inhibited the release of nitric oxide (NO) in RAW264.7 cells stimulated with lipopolysaccharide (LPS). Western blot analysis revealed that fucoidan suppressed the LPS-induced expression of the inducible nitric oxide synthase (iNOS) gene. Moreover, the activation of both nuclear factor-{kappa}B (NF-{kappa}B) and activator protein 1 (AP-1) are key steps in the transcriptional activation of the iNOS gene. Here, it was revealed that fucoidan selectively suppressed AP-1 activation, and that the activation of AP-1 appears to be essential for the induction of iNOS in activated macrophages. This inhibitory effect on AP-1 activation by fucoidan might be associated with its NO blocking and anti-inflammatory effects.

Yang, Jin Won [College of Pharmacy, Chosun University, Gwangju (Korea, Republic of); Yoon, Se Young [College of Pharmacy, Chosun University, Gwangju (Korea, Republic of); LG Household and Healthcare Ltd., Research Park, Daejeon (Korea, Republic of); Oh, Soo Jin [College of Pharmacy and Research Center for Transgenic Cloned Pigs, Chungnam National University, Daejeon (Korea, Republic of); Kim, Sang Kyum [College of Pharmacy and Research Center for Transgenic Cloned Pigs, Chungnam National University, Daejeon (Korea, Republic of); Kang, Keon Wook [College of Pharmacy, Chosun University, Gwangju (Korea, Republic of)]. E-mail: kwkang@chosun.ac.kr

2006-07-21

180

Enhancement of lipopolysaccharide-induced nitric oxide and interleukin-6 production by PEGylated gold nanoparticles in RAW264.7 cells  

NASA Astrophysics Data System (ADS)

While the immunogenicity and cytotoxicity of gold nanoparticles (AuNPs) are noted by many researchers, the mechanisms by which AuNPs exert these effects are poorly understood. In this study, we investigated the effects of polyethylene glycolylated AuNPs (PEG@AuNPs) on lipopolysaccharide (LPS)-induced nitric oxide (NO) and interleukin-6 (IL-6) production and the associated molecular mechanism in RAW264.7 cells. The results showed that PEG@AuNPs were internalized more quickly by LPS-activated RAW264.7 cells than unstimulated cells, and they reached saturation within 24 hours. PEG@AuNPs enhanced LPS-induced production of NO and IL-6 and inducible nitric oxide synthase (iNOS) expression in RAW264.7 cells, partially by activating p38 mitogen-activated protein kinases (p38 MAPK) and nuclear factor-kappaB pathways. In addition, the p38 MAPK inhibitor SB203580 attenuated PEG@AuNP-enhanced LPS-induced NO production and iNOS expression. Overproduction of NO and IL-6 is known to be closely correlated with the pathology of many diseases and inflammations. Thus, it is speculated that the highly biocompatible gold nanoparticles can induce immunotoxicity due to their potency to stimulate macrophages to release aberrant or excessive pro-inflammatory mediators.While the immunogenicity and cytotoxicity of gold nanoparticles (AuNPs) are noted by many researchers, the mechanisms by which AuNPs exert these effects are poorly understood. In this study, we investigated the effects of polyethylene glycolylated AuNPs (PEG@AuNPs) on lipopolysaccharide (LPS)-induced nitric oxide (NO) and interleukin-6 (IL-6) production and the associated molecular mechanism in RAW264.7 cells. The results showed that PEG@AuNPs were internalized more quickly by LPS-activated RAW264.7 cells than unstimulated cells, and they reached saturation within 24 hours. PEG@AuNPs enhanced LPS-induced production of NO and IL-6 and inducible nitric oxide synthase (iNOS) expression in RAW264.7 cells, partially by activating p38 mitogen-activated protein kinases (p38 MAPK) and nuclear factor-kappaB pathways. In addition, the p38 MAPK inhibitor SB203580 attenuated PEG@AuNP-enhanced LPS-induced NO production and iNOS expression. Overproduction of NO and IL-6 is known to be closely correlated with the pathology of many diseases and inflammations. Thus, it is speculated that the highly biocompatible gold nanoparticles can induce immunotoxicity due to their potency to stimulate macrophages to release aberrant or excessive pro-inflammatory mediators. Electronic supplementary information (ESI) available. See DOI: 10.1039/c2nr31355c

Liu, Zhimin; Li, Wenqing; Wang, Feng; Sun, Chunyang; Wang, Lu; Wang, Jun; Sun, Fei

2012-10-01

181

Cryptomeria japonica Essential Oil Inhibits the Growth of Drug-Resistant Skin Pathogens and LPS-Induced Nitric Oxide and Pro-Inflammatory Cytokine Production  

Microsoft Academic Search

In this study, the chemical composition of Cryptomeria japonica essential oil (CJE) was analyzed and its biological activities were tested. CJE was obtained by steam distillation from leaves collected from Jeju Island and analyzed by gas chromatography (GC)-flame ionization detection (FID) and GC-MS. Kaurene (17.20 %), elemol (10.88 %), (-eudesmol (9.41 %), and sabinene (8.86 %) were the major compo-

WEON-JONG YOON; SANG-SUK KIM; TAE-HEON OH; NAM HO LEE; CHANG-GU HYUN

182

Rosmarinic Acid in Prunella vulgaris Ethanol Extract Inhibits LPS-induced Prostaglandin E2 and Nitric Oxide in RAW264.7 Mouse Macrophages  

Technology Transfer Automated Retrieval System (TEKTRAN)

Prunella vulgaris has been used therapeutically for inflammation related conditions for centuries, but systematic studies of its anti-inflammatory activity are lacking and no specific active components have been identified. In this study, water and ethanol extracts of four P. vulgaris accessions we...

183

Fimasartan, anti-hypertension drug, suppressed inducible nitric oxide synthase expressions via nuclear factor-kappa B and activator protein-1 inactivation.  

PubMed

Since inhibition of angiotensin II type 1 (AT1) receptor reduces chronic inflammation associated with hypertension, we evaluated the anti-inflammatory potential and the underlying mechanism of fimasartan, a Korean Food and Drug Administration approved anti-hypertension drug, in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. Fimasartan suppressed the expressions of inducible nitric oxide synthase (iNOS) by down-regulating its transcription, and subsequently inhibited the productions of nitric oxide (NO). In addition, fimasartan attenuated LPS-induced transcriptional and DNA-binding activities of nuclear factor-kappa B (NF-?B) and activator protein-1 (AP-1). These reductions were accompanied by parallel reductions in the nuclear translocation of NF-?B and AP-1. Taken together, our data suggest that fimasartan down-regulates the expression of the iNOS in macrophages via NF-?B and AP-1 inactivation. PMID:23449332

Ryu, Suran; Shin, Ji-Sun; Cho, Young-Wuk; Kim, Hyoung Kook; Paik, Soo Heui; Lee, Joo Han; Chi, Yong Ha; Kim, Ji Han; Kim, Je Hak; Lee, Kyung-Tae

2013-01-01

184

Sesamol down-regulates the lipopolysaccharide-induced inflammatory response by inhibiting nuclear factor-kappa B activation.  

PubMed

We examined the effects of sesamol on the lipopolysaccharide (LPS)-induced inflammatory response. Sesamol inhibited serum tumor necrosis factor (TNF)-?, interleukin (IL)-1? and nitrite production in LPS-treated mice, and inhibited LPS-induced inducible nitric oxide synthase expression in mouse leukocytes. Sesamol also down-regulated TNF-?, IL-1?, and nitrite production as well as inducible nitric oxide synthase expression in LPS-treated RAW 264.7 cells. Further, sesamol inhibited LPS-induced nuclear factor (NF)-?B translocation and inhibitor (I)?B-? phosphorylation in RAW 264.7 cells. By inhibiting TNF-?, IL-1?, and nitrite levels, and interfering with the NF?B pathway, sesamol down-regulated the LPS-initiated inflammatory response. PMID:19939906

Chu, Pei-Yi; Hsu, Dur-Zong; Hsu, Pin-Yen; Liu, Ming-Yie

2010-10-01

185

Anti-angiogenic and inhibitory activity on inducible nitric oxide production of the mushroom Ganoderma lucidum  

Microsoft Academic Search

Fresh fruit bodies of Ganoderma lucidum were extracted with 70% ethanol at room temperature. The extract (GL) showed significant anti-angiogenic activity, which was detected using a chick embryo chorioallantoic membrane assay. GL significantly inhibited LPS-induced NO production in RAW 264.7 macrophages. These results support the anti-tumor effect of Ganoderma lucidum.

Yun Seon Song; Sun-Hyoung Kim; Jae-Hoon Sa; Changbae Jin; Chang-Jin Lim; Eun-Hee Park

2004-01-01

186

A molecular pharmacology study into the anti-inflammatory actions of Euphorbia hirta L. on the LPS-induced RAW 264.7 cells through selective iNOS protein inhibition  

Microsoft Academic Search

Euphorbia hirta L. has been widely used in India and Chinese society. The molecular pharmacology basis of its anti-inflammatory effect is\\u000a revealed in this work. The ethanol extract of Euphorbia hirta L. (Eh) and its active component were studied in lipopolysaccharide (LPS)-activated macrophage cells (RAW 264.7) as an established\\u000a inflammation model. After activation, nitric oxide (NO) production and expression of

Mei-Fen Shih; Yih-Dih Cheng; Chia-Rui Shen; Jong-Yuh Cherng

2010-01-01

187

Bauer ketones 23 and 24 from Echinacea paradoxa var. paradoxa inhibit lipopolysaccharide-induced nitric oxide, prostaglandin E2 and cytokines in RAW264.7 mouse macrophages.  

PubMed

Among the nine Echinacea species, E. purpurea, E. angustifolia and E. pallida, have been widely used to treat the common cold, flu and other infections. In this study, ethanol extracts of these three Echinacea species and E. paradoxa, including its typical variety, E. paradoxa var. paradoxa, were screened in lipopolysaccharide (LPS)-stimulated macrophage cells to assess potential anti-inflammatory activity. E. paradoxa var. paradoxa, rich in polyenes/polyacetylenes, was an especially efficient inhibitor of LPS-induced production of nitric oxide (NO), prostaglandin E2 (PGE2), interleukin-1 beta (IL-1?) and interleukin-6 (IL-6) by 46%, 32%, 53% and 26%, respectively, when tested at 20 ?g/ml in comparison to DMSO control. By bioactivity-guided fractionation, pentadeca-8Z-ene-11, 13-diyn-2-one (Bauer ketone 23) and pentadeca-8Z, 13Z-dien-11-yn-2-one (Bauer ketone 24) from E. paradoxa var. paradoxa were found primarily responsible for inhibitory effects on NO and PGE2 production. Moreover, Bauer ketone 24 was the major contributor to inhibition of inflammatory cytokine production in LPS-induced mouse macrophage cells. These results provide a rationale for exploring the medicinal effects of the Bauer ketone-rich taxon, E. paradoxa var. paradoxa, and confirm the anti-inflammatory properties of Bauer ketones 23 and 24. PMID:22133644

Zhang, Xiaozhu; Rizshsky, Ludmila; Hauck, Catherine; Qu, Luping; Widrlechner, Mark P; Nikolau, Basil J; Murphy, Patricia A; Birt, Diane F

2012-02-01

188

Bauer Ketones 23 and 24 from Echinacea paradoxa var. paradoxa Inhibit Lipopolysaccharide-induced Nitric Oxide, Prostaglandin E2 and Cytokines in RAW 264.7 Mouse Macrophages  

PubMed Central

Among the nine Echinacea species, E. purpurea, E. angustifolia and E. pallida, have been widely used to treat the common cold, flu and other infections. In our study, ethanol extracts of these three Echinacea species and E. paradoxa, including its typical variety, E. paradoxa var. paradoxa, were screened in lipopolysaccharide (LPS)-stimulated macrophage cells to assess potential anti-inflammatory activity. Echinacea paradoxa var. paradoxa, rich in polyenes/polyacetylenes, was an especially efficient inhibitor of LPS-induced production of nitric oxide (NO), prostaglandin E2 (PGE2), interleukin-1 beta (IL-1?) and interleukin-6 (IL-6) by 46%, 32%, 53% and 26%, respectively, when tested at 20 ?g/ml in comparison to DMSO control. By bioactivity-guided fractionation, pentadeca-8Z-ene-11, 13-diyn-2-one (Bauer ketones 23, compound 1) and pentadeca-8Z, 13Z-dien-11-yn-2-one (Bauer ketone 24, compound 2) from E. paradoxa var. paradoxa were found primarily responsible for inhibitory effects on NO and PGE2 production. Moreover, Bauer ketone 24 (compound 2) was the major contributor to inhibition of inflammatory cytokine production in LPS-induced mouse macrophage cells. These results provide a rationale for exploring the medicinal effects of the Bauer ketone-rich taxon, E. paradoxa var. paradoxa, and confirm the anti-inflammatory properties of Bauer ketones 23 and 24. PMID:22133644

Zhang, Xiaozhu; Rizshsky, Ludmila; Hauck, Catherine; Qu, Luping; Widrlechner, Mark P.; Nikolau, Basil J.; Murphy, Patricia A.; Birt, Diane F.

2011-01-01

189

Regulatory role of nitric oxide in lipopolysaccharides-triggered plant innate immunity.  

PubMed

Recent studies have suggested that lipopolysaccharides (LPS) induce nitric oxide (NO) production and defense gene expression in plants. Our current work investigated the signaling mechanism of NO and the role of NONEXPRESSOR OF PATHOGENESIS-RELATED GENES1 (NPR1) in LPS-induced innate immunity of Arabidopsis (Arabidopsis thaliana). We have provided evidence that LPS-elicited NO generation as well as increased antioxidant enzyme activities capable of maintaining the redox state could be important to protect plants against oxidative damage from pathogen attack. In addition, LPS-activated defense responses, including callose deposition and defense-related gene expression, are regulated through an NPR1-dependent signaling pathway. Our results contribute to elucidation of the signaling mechanism of NO and highlight an important role of NPR1 in modulating LPS-triggered innate immunity in plants. However, further research is necessary to clarify the cross-talk between mitochondria and NO on activating LPS-induced defense responses, and the regulatory mechanism of NO in LPS-induced innate immunity needs further improvement. PMID:23221762

Sun, Aizhen; Li, Zhe

2013-01-01

190

Aminoguanidine selectively inhibits inducible nitric oxide synthase.  

PubMed Central

1. Endotoxin induces nitric oxide synthase in vascular tissue, including rat main pulmonary artery. Currently available agents that cause inhibition of nitric oxide synthase are relatively non-selective between the constitutive and inducible forms of the enzyme. 2. Aminoguanidine caused a dose-dependent increase in phenylephrine-induced tension in intact and endothelium-denuded pulmonary artery rings from endotoxin-treated rats, but had no effect on sham-treated controls. 3. Contraction caused by aminoguanidine in endothelium-denuded vessels from endotoxin-treated rats was unaffected by indomethacin (10 microM), and by cimetidine and mepyramine (both 10 microM), excluding an effect of aminoguanidine mediated by arachidonic acid metabolites or histamine. 4. Contraction caused by aminoguanidine in endothelium-denuded vessels from endotoxin-treated rats was abolished by L-arginine (2 mM) and L-NG-monomethyl arginine (300 microM), but unaffected by D-arginine and D-NG-monomethyl arginine, suggesting that its action is mediated by the L-arginine/nitric oxide pathway.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7507781

Griffiths, M. J.; Messent, M.; MacAllister, R. J.; Evans, T. W.

1993-01-01

191

Endotoxin-induced nitric oxide production rescues airway growth and maturation in atrophic fetal rat lung explants  

SciTech Connect

Inflammation induces premature maturation of the fetal lung but the signals causing this effect remain unclear. We determined if nitric oxide (NO) synthesis, evoked by Escherichia coli lipopolysaccharide (LPS, 2 {mu}g ml{sup -1}), participated in this process. Fetal rat lung airway surface complexity rose 2.5-fold over 96 h in response to LPS and was associated with increased iNOS protein expression and activity. iNOS inhibition by N6-(1-iminoethyl)-L-lysine-2HCl (L-NIL) abolished this and induced airway atrophy similar to untreated explants. Surfactant protein-C (SP-C) expression was also induced by LPS and abolished by L-NIL. As TGF{beta} suppresses iNOS activity, we determined if feedback regulation modulated NO-dependent maturation. LPS induced TGF{beta}1 release and SMAD4 nuclear translocation 96 h after treatment. Treatment of explants with a blocking antibody against TGF{beta}1 sustained NO production and airway morphogenesis whereas recombinant TGF{beta}1 antagonized these effects. Feedback regulation of NO synthesis by TGF{beta} may, thus, modulate airway branching and maturation of the fetal lung.

Rae, C. [Division of Maternal and Child Health Sciences, Ninewells Hospital and Medical School, University of Dundee, Dundee, DD1 9SY Scotland (United Kingdom); Cherry, J.I. [Division of Maternal and Child Health Sciences, Ninewells Hospital and Medical School, University of Dundee, Dundee, DD1 9SY Scotland (United Kingdom); Land, F.M. [Division of Maternal and Child Health Sciences, Ninewells Hospital and Medical School, University of Dundee, Dundee, DD1 9SY Scotland (United Kingdom); Land, S.C. [Division of Maternal and Child Health Sciences, Ninewells Hospital and Medical School, University of Dundee, Dundee, DD1 9SY Scotland (United Kingdom)]. E-mail: s.c.land@dundee.ac.uk

2006-10-13

192

Lipopolysaccharide and cytokines induce nitric oxide synthase and produce nitric oxide in cultured normal human melanocytes  

Microsoft Academic Search

\\u000a Abstract The role of nitric oxide in normal and pathological conditions of human skin is still poorly understood. In this study we\\u000a have demonstrated by immunobloting the expression of an inducible nitric oxide synthase isoform (iNOS) in cultured normal\\u000a human melanocytes treated with bacterial lipopolysaccharide, tumor necrosis factor-· and interferon-Á. Nitric oxide was also\\u000a detected in the culture medium and

Irene Machado Rocha; Lidia Andreu Guillo

2001-01-01

193

Atypical “seizure-like” activity in cortical reverberating networks in vitro can be caused by LPS-induced inflammation: a multi-electrode array study from a hundred neurons  

PubMed Central

We show here that a mild sterile inflammation induced by the endotoxin lipopolysaccharide (LPS), in a neuron/astrocyte/microglial cortical network, modulates neuronal excitability and can initiate long-duration burst events resembling epileptiform seizures, a recognized feature of various central nervous neurodegenerative, neurological and acute systemic diseases associated with neuroinflammation. To study this action, we simultaneously analyzed the reverberating bursting activity of a hundred neurons by using in vitro multi-electrode array methods. ?5 h after LPS application, we observed a net increase in the average number of spikes elicited in engaged cells and within each burst, but no changes neither in spike waveforms nor in burst rate. This effect was characterized by a slow, twofold exponential increase of the burst duration and the appearance of rarely occurring long burst events that were never seen during control recordings. These changes and the time-course of microglia-released proinflammatory cytokine, tumor necrosis factor-alpha (TNF-?), were blocked by pre-treatment with 50 nM minocycline, an established anti-inflammatory agent which was inactive when applied alone. Assay experiments also revealed that application of 60 pM exogenous TNF-? after 12–15 h, produced non-washable changes of neuronal excitability, completely different from those induced by LPS, suggesting that TNF-? release alone was not responsible for our observed findings. Our results indicate that the link between neuroinflammation and hyperexcitability can be unveiled by studying the long-term activity of in vitro neuronal/astrocyte/microglial networks. PMID:25404893

Gullo, Francesca; Amadeo, Alida; Donvito, Giulia; Lecchi, Marzia; Costa, Barbara; Constanti, Andrew; Wanke, Enzo

2014-01-01

194

Effect of methanolic extract of Asparagus racemosus Willd. on lipopolysaccharide induced-oxidative stress in rats.  

PubMed

Lipopolysaccharide (LPS) induced oxidative stress and impairment of normal physiological function generally categorized by increased anxiety and reduced mobility. Therefore, the present study was to find out the effect Methanolic extract of Asparagus racemosus (MEAR ) in lipopolysaccharide (LPS)-induced oxidative stress in rats . LPS-induced oxidative stress in rats was measured by locomotor activity by photoactometer test, anxiety with elevated plus maze test and also studied the oxidative stress markers, nitric oxide and cytokines. The obtained data shows that LPS markedly exhausted (p<0.001) brain- reduced glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT) significantly increased (p<0.001) the level of malondialdehyde (MDA), nitric oxide and the activity of cytokines in the brain. MEAR supplementation resulted in normalization of brain GSH and CAT and SOD and decreases in the levels of MDA with reduction of nitric oxide and cytokines in the brain. The action of the extract at dose of 200 mg/kg was almost similar to the standard drug, quercetin (100mg/kg, p.o.). These present study conclude that MEAR administration significantly (P<0.05) reduced LPS- induced oxidative-stress and intensely suggest that Asparagus racemosus Willd. is a functionally newer type of cerebroprotective agent. PMID:25730806

Ahmad, Mohammad Parwez; Hussain, Arshad; Siddiqui, Hefazat Hussain; Wahab, Shadma; Adak, Manoranjan

2015-03-01

195

Borrelia burgdorferi and Escherichia coli lipopolysaccharides induce nitric oxide and interleukin-6 production in cultured rat brain cells.  

PubMed

Borrelia burgdorferi, the spirochetal agent of Lyme disease, infects the central nervous system (CNS), but the factors that mediate inflammation and neurologic dysfunction are not known. Sonicated B. burgdorferi stimulated in a concentration-dependent manner the production of nitric oxide (NO) in glial-enriched primary cultures of neonatal rat brain cells via induction of NO synthase activity. Lipopolysaccharide (LPS) of Escherichia coli also stimulated nitrite accumulation in a concentration-dependent manner. Stimulation of NO production by B. burgdorferi sonicate and E. coli LPS was associated with increased levels of mRNA coding for the cytokine-inducible form of NO synthase. B. burgdorferi sonicate also stimulated release of interleukin-6, with a concentration-response relationship similar to that for its stimulation of nitrite production, as did E. coli LPS. A competitive antagonist of E. coli LPS, Rhodopseudomonas sphaeroides lipid A, inhibited LPS-induced stimulation of NO synthase activity but markedly potentiated that of B. burgdorferi, indicating that the initial triggering mechanism of B. burgdorferi is distinct from that of E. coli LPS. Induction of NO synthase by bacterial agents within the brain may represent a common pathway of CNS inflammation and neurotoxicity. PMID:7513330

Tatro, J B; Romero, L I; Beasley, D; Steere, A C; Reichlin, S

1994-05-01

196

Role of Nitric Oxide in Cocaine-Induced Acute Hypertension  

Microsoft Academic Search

Cocaine causes acute hypertension by blocking catecholamine reuptake. There is evidence that it also impairs the peripheral endothelial nitric oxide system, which is normally vasodilatory. We further explored the role of nitric oxide in cocaine-induced vasoconstriction in anesthetized rats, and in vitro by using isolated carotid artery segments. Cocaine administered intravenously in rats increased mean arterial pressure by 30 to

Wa Mo; Ashok K. Singh; Jose A. L. Arruda; George Dunea

1998-01-01

197

Inhibitory effects of salidroside on nitric oxide and prostaglandin E? production in lipopolysaccharide-stimulated RAW 264.7 macrophages.  

PubMed

The aim of this study was to evaluate the effect of salidroside on lipopolysaccharide (LPS)-induced nitric oxide (NO) and prostaglandin E? (PGE?) production in RAW 264.7 macrophages and related anti-inflammatory mechanism. PGE? production was measured by enzyme-linked immunosorbent assay (ELISA); NO production was tested by Griess reagent. Inducible nitric oxidesynthase (iNOS) and COX-2 were determined by RT-PCR and Western blot analysis; I?B and P-I?B protein express were detected by Western blot analysis; cytosolic free Ca²? ([Ca²?](i)) was measured by a fluorescent microscope. The data showed salidroside inhibited LPS-induced NO and PGE? production and reduced iNOS and COX-2 protein expression in RAW 264.7 macrophages. Consistent with these observations, salidroside inhibited LPS-induced cytosolic free Ca²? concentration ([Ca²?](i)) elevation. In addition, we further investigated signal transduction mechanisms and found that the activation of NF-?B was suppressed by salidroside in a dose-dependent manner. These results suggest that salidroside suppresses NO and PGE? production by inhibiting iNOS and COX-2 protein expression, level of [Ca²?](i), and activation of NF-?B signal transduction pathway. PMID:24180550

Song, Bocui; Huang, Guoren; Xiong, Ying; Liu, Jingbo; Xu, Linli; Wang, Zhenning; Li, Gen; Lu, Jing; Guan, Shuang

2013-11-01

198

Di- and sesqui-terpenoids isolated from the pods of Sindora sumatrana and their potential to inhibit lipopolysaccharide-induced nitric oxide production.  

PubMed

Activity-guided fractionation of the n-hexane and CHCl3-soluble fractions of Sindora sumatrana using a bioassay based on the inhibition of lipopolysaccharide (LPS)-induced nitric oxide (NO) production by inducible nitric oxide synthase (iNOS) in murine macrophage RAW 264.7 cells led to the isolation of the known compound, (+)-7beta-acetoxy-15,16-epoxy-3,13(16),14-clerodatriene-18-oic acid (2) as an active constituent. In addition, a new trans-clerodane diterpenoid, (+)-2-oxokolavenic acid (1), together with six known compounds, (+)-3,13-clerodadiene-16,15-olide-18-oic acid (3), (+)-7beta-acetoxy-3,13-clerodadiene-16,15-olide-18-oic acid (4), (+)-7beta-acetoxy-16-hydroxy-3,13-clerodadiene-16,15-olide-18-oic acid (5), beta-caryophyllene oxide (6), clovane-2beta,9beta-diol (7), and caryolane-1,9beta-diol (8) were isolated and found to be inactive. The structure of compound 1 was determined using physical and spectroscopic methods such as 1D and 2D-NMR experiments. The known compounds 2-8 were identified by the spectroscopic data and by comparison with the published values. Of eight isolates (1-8), only compound 2 exhibited an iNOS inhibitory activity with IC50 value of 51.6 microM. PMID:15089033

Jang, Dae Sik; Min, Hye-Young; Jeong, Yeon-Hee; Lee, Sang Kook; Seo, Eun-Kyoung

2004-03-01

199

Polar lipids from the marine macroalga Palmaria palmata inhibit lipopolysaccharide-induced nitric oxide production in RAW264.7 macrophage cells.  

PubMed

The EtOAc soluble fraction of a MeOH/CHCl3 extract of Palmaria palmata showed strong nitric oxide (NO) inhibitory activity against lipopolysaccharide (LPS)-induced NO production in murine RAW264.7 cells. NO inhibition-guided isolation led to identification of three new polar lipids including a sulfoquinovosyl diacylglycerol (SQDG) (2S)-1-O-eicosapentaenoyl-2-O-myristoyl-3-O-(6-sulfo-?-D-quinovopyranosyl)-glycerol (1) and two phosphatidylglycerols, 1-O-eicosapentaenoyl-2-O-trans-3-hexadecenoyl-3-phospho-(1'-glycerol)-glycerol (3) and 1-O-eicosapentaenoyl-2-O-palmitoyl-3-phospho-(1'-glycerol)-glycerol (4) from the EtOAc fraction. Seven known lipids were also isolated including a SQDG (2), a phospholipid (5) and five galactolipids (6-10). Structures of the isolated lipids were elucidated by spectral analyses. The isolated SQDGs, phosphatidylglycerols and phospholipid possessed strong and dose-dependent NO inhibitory activity compared to N(G)-methyl-L-arginine acetate salt (L-NMMA), a well-known NO inhibitor used as a positive control. Further study suggested that these polar lipids suppressed NO production through down-regulation of inducible nitric oxide synthase (iNOS). PMID:24569177

Banskota, Arjun H; Stefanova, Roumiana; Sperker, Sandra; Lall, Santosh P; Craigie, James S; Hafting, Jeff T; Critchley, Alan T

2014-05-01

200

Methanol extract of Ficus leaf inhibits the production of nitric oxide and proinflammatory cytokines in LPS-stimulated microglia via the MAPK pathway.  

PubMed

Excessive production of inflammatory mediators, nitric oxide (NO) and proinflammatory cytokines from activated microglia has been implicated in neurodegeneration in human brain diseases. Recently, it seems possible that treatment with antiinflammatory agents, including Oriental medicinal plants, might delay the progression of neurodegeneration through the inhibition of microglial activation. The present study evaluated the effect of a methanol extract of Ficus religiosa leaf (MFL) on lipopolysaccharide (LPS)-induced production of NO and proinflammatory cytokines, such as tumor necrosis factor-alpha (TNF-alpha), interleukin-beta (IL-1beta) and IL-6 in BV-2 cells, a mouse microglial line. MFL inhibited LPS-induced production of NO and proinflammatory cytokines in a dose-dependent manner. MFL also attenuated the expression of mRNA and proteins of inducible nitric oxide synthase (iNOS) and proinflammatory cytokines, suggesting the blockage of transcription levels, respectively. The molecular mechanism of MFL-mediated attenuation underlies the down-regulation of the extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MAPK) signaling pathway, and suppresses the nuclear factor kappaB (NF-kappaB) activation. The results suggest that MFL exhibits antiinflammatory properties in LPS-induced activation of BV2 microglial cells, and that might have a therapeutic potential for various neurodegenerative diseases. PMID:18546149

Jung, Hyo Won; Son, Hye Young; Minh, Chau Van; Kim, Young Ho; Park, Yong-Ki

2008-08-01

201

Ganoderma lucidum inhibits inducible nitric oxide synthase expression in macrophages  

Microsoft Academic Search

Nitric oxide (NO) is a principal mediator in many physiological and pathological processes. Overproduction of NO via the inducible nitric oxide synthase (iNOS) has cytotoxic effect through the formation of peroxynitrite with superoxide anion. The iNOS is mainly expressed in macrophages and is able to produce large amount of NO. The expression of iNOS is mainly regulated at the transcriptional

Connie W. H. Woo; Ricky Y. K. Man; Yaw L. Siow; Patrick C. Choy; Eric W. Y. Wan; Chak S. Lau; Karmin O

2005-01-01

202

Inhibitory effect of resveratrol dimerized derivatives on nitric oxide production in lipopolysaccharide-induced RAW 264.7 cells.  

PubMed

Four types of resveratrol dimerized analogues were synthesized and evaluated in vitro on LPS-induced NO production in RAW 264.7 cells. The results showed that several compounds, especially those containing 1,2-diphenyl-2,3-dihydro-1H-indene core (type I), exhibited good inhibitory activities. Among 25 analogues, 12b showed a significant inhibitory activity (49% NO production at 10 ?M, IC50=3.38 ?M). Further study revealed that compound 12b could suppress LPS-induced iNOS expression, NO production, and IL-1? release in a concentration-dependently manner. The mechanism of action (MOA) involved for its anti-inflammatory responses was through signaling pathways of p38 MAPK and JNK1/2, but not ERK1/2. PMID:23791078

Zhong, Chen; Liu, Xin-Hua; Chang, Jun; Yu, Jian-Ming; Sun, Xun

2013-08-01

203

Nitric Oxide Sustains IL-1? Expression in Human Dendritic Cells Enhancing Their Capacity to Induce IL-17–Producing T-Cells  

PubMed Central

The role played by lung dendritic cells (DCs) which are influenced by external antigens and by their redox state in controlling inflammation is unclear. We studied the role played by nitric oxide (NO) in DC maturation and function. Human DCs were stimulated with a long-acting NO donor, DPTA NONOate, prior to exposure to lipopolysaccharide (LPS). Dose-and time-dependent experiments were performed with DCs with the aim of measuring the release and gene expression of inflammatory cytokines capable of modifying T-cell differentiation, towardsTh1, Th2 and Th17 cells. NO changed the pattern of cytokine release by LPS-matured DCs, dependent on the concentration of NO, as well as on the timing of its addition to the cells during maturation. Addition of NO before LPS-induced maturation strongly inhibited the release of IL-12, while increasing the expression and release of IL-23, IL-1? and IL-6, which are all involved in Th17 polarization. Indeed, DCs treated with NO efficiently induced the release of IL-17 by T-cells through IL-1?. Our work highlights the important role that NO may play in sustaining inflammation during an infection through the preferential differentiation of the Th17 lineage. PMID:25853810

Obregon, Carolina; Graf, Lukas; Chung, Kian Fan; Cesson, Valerie; Nicod, Laurent P.

2015-01-01

204

Avicularin Inhibits Lipopolysaccharide-Induced Inflammatory Response by Suppressing ERK Phosphorylation in RAW 264.7 Macrophages  

PubMed Central

suppresAvicularin, quercetin-3-?-L-arabinofuranoside, has been reported to possess diverse pharmacological properties such as anti-inflammatory and anti-infectious effects. However, the underlying mechanism by which avicularin exerts its anti-inflammatory activity has not been clearly demonstrated. This study aimed to elucidate the anti-inflammatory mechanism of avicularin in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cells. Avicularin significantly inhibited LPS-induced excessive production of pro-inflammatory mediators such as nitric oxide (NO) and PGE2 and the protein levels of iNOS and COX-2, which are responsible for the production of NO and PGE2, respectively. Avicularin also suppressed LPS-induced overproduction of pro-inflammatory cytokine IL-1?. Furthermore, avicularin significantly suppressed LPS-induced degradation of I?B, which retains NF-?B in the cytoplasm, consequently inhibiting the transcription of pro-inflammatory genes by NF-?B in the nucleus. To understand the underlying signaling mechanism of anti-inflammatory activity of avicularin, involvement of multiple kinases was examined. Avicularin significantly attenuated LPS-induced activation of ERK signaling pathway in a concentration-dependent manner. Taken together, the present study clearly demonstrates that avicularin exhibits anti-inflammatory activity through the suppression of ERK signaling pathway in LPS-stimulated RAW 264.7 macrophage cells. PMID:24009846

Vo, Van Anh; Lee, Jae-Won; Chang, Ji-Eun; Kim, Ji-Young; Kim, Nam-Ho; Lee, Hee Jae; Kim, Sung-Soo; Chun, Wanjoo; Kwon, Yong-Soo

2012-01-01

205

Plumbagin, a vitamin K3 analogue, abrogates lipopolysaccharide-induced oxidative stress, inflammation and endotoxic shock via NF-?B suppression.  

PubMed

Plumbagin has been reported to modulate cellular redox status and suppress NF-?B. In the present study, we investigated the effect of plumbagin on lipopolysaccharide (LPS)-induced endotoxic shock, oxidative stress and inflammatory parameters in vitro and in vivo. Plumbagin inhibited LPS-induced nitric oxide, TNF-?, IL-6 and prostaglandin-E2 production in a concentration-dependent manner in RAW 264.7 cells without inducing any cell death. Plumbagin modulated cellular redox status in RAW cells. Plumbagin treatment significantly reduced MAPkinase and NF-?B activation in macrophages. Plumbagin prevented mice from endotoxic shock-associated mortality and decreased serum levels of pro-inflammatory markers. Plumbagin administration ameliorated LPS-induced oxidative stress in peritoneal macrophages and splenocytes. Plumbagin also attenuated endotoxic shock-associated changes in liver and lung histopathology and decreased the activation of ERK and NF-?B in liver. These findings demonstrate the efficacy of plumbagin in preventing LPS-induced endotoxemia and also provide mechanistic insights into the anti-inflammatory effects of plumbagin. PMID:24234154

Checker, Rahul; Patwardhan, Raghavendra S; Sharma, Deepak; Menon, Jisha; Thoh, Maikho; Sandur, Santosh K; Sainis, Krishna B; Poduval, T B

2014-04-01

206

UV Induced Oxidation of Nitric Oxide  

NASA Technical Reports Server (NTRS)

Nitric oxide in a gaseous stream is converted to nitrogen dioxide using oxidizing species generated at least in part using in situ UV radiation sources. The sources of the oxidizing species include oxygen and/or hydrogen peroxide. The oxygen may be a component of the gaseous stream or added to the gaseous stream, preferably near a UV radiation source, and is converted to ozone by the UV irradiation. The hydrogen peroxide is decomposed through a combination of vaporization and UV irradiation. The hydrogen peroxide is preferably stored at stable concentration levels, i.e., approximately 50% by volume and increased in concentration in a continuous process preceding vaporization within the flow channel of the gaseous stream and in the presence of the UV radiation sources.

Parrish, Clyde, F. (Inventor); Luecke, Dale E. (Inventor)

2007-01-01

207

Design and Synthesis of 2-Amino-4-methylpyridine Analogues as Inhibitors for Inducible Nitric Oxide Synthase and in vivo Evaluation of [18F]6-(2-Fluoropropyl)-4-methyl-pyridin-2-amine as a Potential PET Tracer for Inducible Nitric Oxide Synthase  

PubMed Central

A series of position-6 substituted 2-amino-4-methylpyridine analogues was synthesized and compounds 9, 18, and 20 were identified as the inhibitors with the greatest potential to serve as PET tracers for imaging inducible nitric oxide synthase (iNOS). [18F]9 was synthesized and evaluated in a mouse model of lipopolysaccharide (LPS)-induced iNOS activation. In vivo biodistribution studies of [18F]9 indicate higher tracer uptake in the lungs of the LPS-treated mice when compared to control mice. Tracer uptake at 60 min post-injection was reduced in a blocking study using a known inhibitor of iNOS. The expression of iNOS was confirmed by Western blot analysis of lung samples from the LPS-treated mice. MicroPET studies also demonstrated accumulation of radiotracer in the lungs of the LPS-treated mice. Taken collectively, these data suggest that [18F]9 shows favorable properties as a PET tracer to image iNOS activation with PET. PMID:19323559

Zhou, Dong; Lee, Hsiaoju; Rothfuss, Justin M.; Chen, Delphine L.; Ponde, Datta E.; Welch, Michael J.; Mach, Robert H.

2009-01-01

208

Diethylcarbamazine (DEC) Does Not Induce Nitric Oxide (NO) Synthesis  

Microsoft Academic Search

Rajan, T. V., Shultz, L. D., Babu, S., Doukas, J., Greiner, D., and Porte, P. 1998. Diethylcarbamazine (DEC) does not induce nitric oxide (NO) synthesis.Experimental Parasitology88, 217–222. Diethylcarbomazine (DEC) was discovered in 1947 as a potent therapeutic agent in lymphatic filariasis and has been a mainstay of antifilarial therapy over the past five decades (R. I. Hewitt,et al., 1947,Journal of

T. V. Rajan; Leonard D. Shultz; Subash Babu; John Doukas; Dale Greiner; Pat Porte

1998-01-01

209

Nitric oxide-induced suspended animation promotes survival during hypoxia  

PubMed Central

Oxygen plays a key role in energy metabolism. However, there are organisms that survive severe shortfalls in oxygen. Drosophila embryos rapidly arrest development upon severe hypoxia and recover upon restoration of oxygen, even days later. Stabilization of the normally unstable engrailed RNA and protein preserved the localized striped pattern of this embryonic patterning gene during 3 days in hypoxia. Severe hypoxia blocked expression of a heat-shock-inducible lacZ transgene. Cyanide, a metabolic poison, did not immediately block gene expression or turnover, arguing against a passive response to energy limitation. In contrast, nitric oxide, a putative hypoxia signal, induced a reversible arrest of development, gene expression and turnover. Reciprocally, a nitric oxide scavenger allowed continued gene expression and turnover during hypoxia, but it reduced hypoxia tolerance. We suggest that hypoxia-induced stasis preserves the status quo of embryonic processes and promotes survival. Our data implicate nitric oxide as a mediator of this response and provide a system in which to investigate its action. PMID:12554658

Teodoro, Rita O.; O’Farrell, Patrick H.

2003-01-01

210

Eupatolide inhibits lipopolysaccharide-induced COX-2 and iNOS expression in RAW264.7 cells by inducing proteasomal degradation of TRAF6.  

PubMed

Inula britannica is a traditional medicinal plant used to treat bronchitis, digestive disorders, and inflammation in Eastern Asia. Here, we identified eupatolide, a sesquiterpene lactone from I. britannica, as an inhibitor of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) expression. Eupatolide inhibited the production of nitric oxide (NO) and prostaglandin E(2) (PGE(2)) as well as iNOS and COX-2 protein expression in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Eupatolide dose-dependently decreased the mRNA levels and the promoter activities of COX-2 and iNOS in LPS-stimulated RAW264.7 cells. Moreover, eupatolide significantly suppressed the LPS-induced expression of nuclear factor-kappa B (NF-kappaB) and activator protein-1 (AP-1) reporter genes. Pretreatment of eupatolide inhibited LPS-induced phosphorylation and degradation of I kappaB alpha, and phosphorylation of RelA/p65 on Ser-536 as well as the activation of mitogen-activated protein kinases (MAPKs) and Akt in LPS-stimulated RAW264.7 cells. Eupatolide induced proteasomal degradation of tumor necrosis factor receptor-associated factor-6 (TRAF6), and subsequently inhibited LPS-induced TRAF6 polyubiquitination. These results suggest that eupatolide blocks LPS-induced COX-2 and iNOS expression at the transcriptional level through inhibiting the signaling pathways such as NF-kappaB and MAPKs via proteasomal degradation of TRAF6. Taken together, eupatolide may be a novel anti-inflammatory agent that induces proteasomal degradation of TRAF6, and a valuable compound for modulating inflammatory conditions. PMID:20353767

Lee, Jongkyu; Tae, Nara; Lee, Jung Joon; Kim, Taeho; Lee, Jeong-Hyung

2010-06-25

211

Inhibition of inducible nitric oxide synthase ameliorates rat lung allograft rejection  

Microsoft Academic Search

Recently, the inducible isoform of nitric oxide synthase has been shown to be an important immunomodulation molecule in allograft rejection. We have observed the production of nitric oxide during rejection and the effect of nitric oxide synthase inhibition on allograft rejection in a rat lung transplant model. Rat left lung allotransplants were performed in two strain combinations: brown Norway–to–F344 (major

Takeshi Shiraishi; Steven R. DeMeester; Neil K. Worrall; Jon H. Ritter; Thomas P. Misko; T. Bruce Ferguson; Joel D. Cooper; G. Alexander Patterson

1995-01-01

212

Cannabidiol reduces lipopolysaccharide-induced vascular changes and inflammation in the mouse brain: an intravital microscopy study  

PubMed Central

Background The phytocannabinoid cannabidiol (CBD) exhibits antioxidant and antiinflammatory properties. The present study was designed to explore its effects in a mouse model of sepsis-related encephalitis by intravenous administration of lipopolysaccharide (LPS). Methods Vascular responses of pial vessels were analyzed by intravital microscopy and inflammatory parameters measured by qRT-PCR. Results CBD prevented LPS-induced arteriolar and venular vasodilation as well as leukocyte margination. In addition, CBD abolished LPS-induced increases in tumor necrosis factor-alpha and cyclooxygenase-2 expression as measured by quantitative real time PCR. The expression of the inducible-nitric oxide synthase was also reduced by CBD. Finally, preservation of Blood Brain Barrier integrity was also associated to the treatment with CBD. Conclusions These data highlight the antiinflammatory and vascular-stabilizing effects of CBD in endotoxic shock and suggest a possible beneficial effect of this natural cannabinoid. PMID:21244691

2011-01-01

213

Lipopolysaccharide induces nitric oxide synthase expression and platelet-activating factor increases nitric oxide production in human fetal membranes in culture  

Microsoft Academic Search

BACKGROUND: Platelet-activating factor and nitric oxide may be involved in the initiation of human labour as inflammatory mediators. The aim of this study was to test whether platelet-activating factor and lipopolysaccharide were able to induce nitric oxide synthase expression and stimulate the production of nitric oxide in human fetal membrane explants in culture. METHODS: Fetal membranes were collected from Caesarean

Gunter Seyffarth; Paul N Nelson; Simon J Dunmore; Nalinda Rodrigo; Damian J Murphy; Ray J Carson

2004-01-01

214

Luteolin Suppresses Inflammatory Mediator Expression by Blocking the Akt/NF?B Pathway in Acute Lung Injury Induced by Lipopolysaccharide in Mice  

PubMed Central

Acute lung injury (ALI), instilled by lipopolysaccharide (LPS), is a severe illness with excessive mortality and has no specific treatment strategy. Luteolin is an anti-inflammatory flavonoid and widely distributed in the plants. Pretreatment with luteolin inhibited LPS-induced histological changes of ALI and lung tissue edema. In addition, LPS-induced inflammatory responses, including increased vascular permeability, tumor necrosis factor (TNF)-? and interleukin (IL)-6 production, and expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), were also reduced by luteolin in a concentration-dependent manner. Furthermore, luteolin suppressed activation of NF?B and its upstream molecular factor, Akt. These results suggest that the protection mechanism of luteolin is by inhibition of NF?B activation possibly via Akt. PMID:22203870

Li, Yi-Ching; Yeh, Chung-Hsin; Yang, Ming-Ling; Kuan, Yu-Hsiang

2012-01-01

215

Macrophage inflammatory protein-1 alpha production in lipopolysaccharide-stimulated human adherent blood mononuclear cells is inhibited by the nitric oxide synthase inhibitor N(G)-monomethyl-L-arginine.  

PubMed

The release of chemokines such as macrophage-inflammatory protein-1 alpha (MIP-1 alpha) from activated macrophages is a crucial step in cell recruitment necessary for establishing local inflammatory responses. To ascertain the importance of the L-arginine/nitric oxide (NO) pathway in LPS-induced MIP-1 alpha release, we stimulated human adherent PBMC with LPS in the presence of the NO synthase inhibitor N(G)-monomethyl-L-arginine (L-NMMA). L-NMMA decreased LPS-induced MIP-1 alpha protein release (45.5% inhibition) and steady state levels of mRNA (48% inhibition) in adherent PBMC. The concentration of L-NMMA for inhibition of MIP-1 alpha release was dependent on the concentration of L-arginine in the cell culture medium, emphasizing the L-arginine-related action of the drug. Most of the MIP-1 alpha release was attributed to the activity of IL-1 and TNF, since coincubation of LPS-stimulated PBMC with IL-1R antagonist and TNF-binding protein abrogated LPS-induced MIP-1 alpha release (by 76.8%). Analysis of cytokine secretion revealed that, in addition to MIP-1 alpha, L-NMMA inhibited the release of mature IL-1 beta (by 70%) and TNF-alpha (by 53%). In contrast, release of macrophage chemoattractant protein-1 was unaffected; IL-10 was augmented (123.4%) by L-NMMA. In the presence of exogenous NO provided by NO donors, LPS-induced MIP-1 alpha release was enhanced. We concluded that endogenous NO acts as a mediator of inflammation. Since IL-10 is a potent anti-inflammatory cytokine, these data also suggest that L-NMMA acts as an anti-inflammatory agent by specifically altering the balance of pro- and anti-inflammatory cytokines released from LPS-stimulated human PBMC. PMID:9366434

Mühl, H; Dinarello, C A

1997-11-15

216

The role of nitric oxide in experimental cerulein induced pancreatitis.  

PubMed Central

An enhanced formation of nitric oxide (NO), due to the induction of inducible nitric oxide synthase (iNOS), has been implicated in the pathogenesis of shock and inflammation, but its role in acute pancreatitis still remains controversial. To clarify the role of NO in acute pancreatitis, the present experiment investigated the expression of iNOS and the effect of NOS inhibition on cerulein-induced pancreatitis in rats. Group I received intraperitoneal (ip) injection of normal saline. Group II received two ip injections of cerulein (20 microgram/kg). Group III received injections of N(G)-nitro-L-arginine methyl ester (L-NAME) (30 mg/kg) with cerulein. Group IV received L-arginine (250 mg/kg) with cerulein and L-NAME. The expression of iNOS in the pancreas was examined by western blot analysis. The plasma concentration of NO metabolites was measured. The severity of pancreatitis was assessed by measuring serum amylase, pancreas water content and histopathological examination. Compared with controls, the cerulein group displayed significantly increased expression of iNOS and raised plasma NO metabolites. Treatment with L-NAME significantly decreased hyperamylasemia, plasma NO level, and the extent of pancreatic injury. Treatment with L-arginine reversed the effects of L-NAME. These findings suggest that an enhanced formation of NO by iNOS plays an important role in the development of acute pancreatitis, and inhibition of NO production has the beneficial effects in reducing pancreas injury. PMID:12923328

Um, Soon Ho; Kwon, Yong Dae; Kim, Chang Duck; Lee, Hong Sik; Jeen, Yoon Tae; Chun, Hoon Jai; Lee, Sang Woo; Choi, Jae Hyun; Ryu, Ho Sang; Hyun, Jin Hai

2003-01-01

217

[Inducible nitric oxide synthase expression and nitric oxide production by monocytes in systemic sclerosis].  

PubMed

We investigated nitric oxide (NO) production and inducible NO synthase (iNOS) expression by cultured peripheral blood mononuclear cells (PBMC) in systemic sclerosis (SSc). Eighteen patients with SSc were compared to two control groups: 16 rheumatoid arthritis patients (RA) and 23 mechanical sciatica patients. The sum of nitrites and nitrates was determined by fluorimetry in sera and spectrophotometry in supernatants. Inducible iNOS was detected in cultured PBMC by immunofluorescence, immunoblot and flow cytometry with or without IL-1 beta + TNF alpha, IL-4 or IFN gamma from day 1 to day 5. NO metabolite concentrations in the plasma were lower in SSc (34.3 mumol/l +/- 2.63 SEM) than in RA (48.3 mumol/l +/- 2.2; p < 0.02) and sciatica (43.3 mumol/l +/- 5.24; p < 0.03) patients. iNOS was detected in cultured monocytes in the 3 groups but induction occurred on day 1 in RA, day 2 in sciatica and only on day 3 in SSc, whatever the stimulus. The concentrations of NO metabolites are decreased in SSc patients and the induction of iNOS in PBMC is delayed. Low levels of NO, a vasodilator, may be involved in vasospasm, which is critical in SSc. This may suggest therapeutic implications. PMID:11501260

Menkès, C J; Allanore, Y; Borderie, D; Hilliquin, P; Hernvann, A; Ekindjian, O; Kahan, A

2001-01-01

218

Inducible Nitric Oxide Synthase Expression in Human Colorectal Cancer  

PubMed Central

To investigate the potential involvement of the nitric oxide (NO) pathway in colorectal carcinogenesis, we correlated the expression and the activity of inducible nitric oxide synthase (iNOS) with the degree of tumor angiogenesis in human colorectal cancer. Tumor samples and adjacent normal mucosa were obtained from 46 surgical specimens. Immunohistochemical expression of iNOS, vascular endothelial growth factor (VEGF), and CD31 was analyzed on paraffin-embedded tissue sections. iNOS activity and cyclic GMP levels were assessed by specific biochemical assays. iNOS protein expression was determined by Western blot analysis. iNOS and VEGF mRNA levels were evaluated using Northern blot analysis. Both iNOS and VEGF expressions correlated significantly with intratumor microvessel density (rs = 0.31, P = 0.02 and rs = 0.67, P < 0.0001, respectively). A significant correlation was also found between iNOS and VEGF expression (P = 0.001). iNOS activity and cyclic GMP production were significantly higher in the cancer specimens than in the normal mucosa (P < 0.0001 and P < 0.0001, respectively), as well as in metastatic tumors than in nonmetastatic ones (P = 0.002 and P = 0.04, respectively). Western and Northern blot analyses confirmed the up-regulation of the iNOS protein and gene in the tumor specimens as compared with normal mucosa. NO seems to play a role in colorectal cancer growth by promoting tumor angiogenesis. PMID:12598314

Cianchi, Fabio; Cortesini, Camillo; Fantappiè, Ornella; Messerini, Luca; Schiavone, Nicola; Vannacci, Alfredo; Nistri, Silvia; Sardi, Iacopo; Baroni, Gianna; Marzocca, Cosimo; Perna, Federico; Mazzanti, Roberto; Bechi, Paolo; Masini, Emanuela

2003-01-01

219

Nitric Oxide Signaling in Hypergravity-Induced Neuronal Plasticity  

NASA Technical Reports Server (NTRS)

The goal of this research project was to identify the neurons and circuits in the vestibular nuclei and nucleus prepositus hypoglossi that utilize nitric oxide (NO) for intercellular signaling during gravity-induced plasticity. This objective was pursued using histochemical and immunocytochemical approaches to localize NO-producing neurons and characterize the fine morphology of the cells in ground-based studies of normal rats, rats adapted to hypergravity, and rats adapted to hypergravity and then re-adapted to the 1G environment. NO-producing neurons were identified and studied using four methodologies: i) immunocytochemistry employing polyclonal antibodies directed against neuronal nitric oxide synthase (nNOS), to provide an indication of the capacity of a cell for NO production; ii) immunocytochemistry employing a monoclonal antibody directed against L-citrulline, to provide an indirect index of the enzyme's activity; iii) histochemistry based on the NADPH-diaphorase reaction, for fuI1 cytological visualization of neurons; and iv) double immunofluorescence to co-localize nNOS and L-citrulline in individual vestibular nuclei (VN) and neurons.

Holstein, Gay R.

2003-01-01

220

The effect of inhaled nitric oxide on the carrageenan-induced paw edema.  

PubMed

Inhaled nitric oxide therapy reaches not only pulmonary vessels, but also other vasculatures, presenting anti-inflammatory effects. Therefore, this study investigated the effects of inhaled nitric oxide on a mice model of carrageenan-induced paw edema. Paw edema was induced in male Swiss mice (20-30 g) by subplantar injection of carrageenan (0.05 ml of a 1% suspension in 0.9% saline). The evaluation of time-course edema (mililiter) was measured by plethysmometry until 12 h following carrageenan administration. Thirty minutes after carrageenan injection, some groups received inhaled nitric oxide (300 ppm at variable doses and times) or Indometacin (INDO 5 mg/Kg, v.o), while others received sildenafil (1 mg/Kg, i.p) or rolipram (3 mg/Kg, i.p.) with or without inhaled nitric oxide. Paws were assessed for edema levels by plethysmometry, mieloperoxidase activity and histological analysis. Inhaled nitric oxide significantly reduced carrageenan-induced paw edema, mieloperoxidase activity and inflammatory infiltrate, although similar results were also observed in sildenafil and rolipram treated groups. In addition, significant effects between inhaled nitric oxide with pharmacological therapy was observed. Inhaled nitric oxide presents anti-inflammatory effects on carrageenan-induce paw edema, as observed through reduced edema, mieloperoxidase activity and neutrophil infiltration, indicating that inhaled nitric oxide therapy goes beyond lung vascular effects. PMID:25070733

Coelho, Carly Faria; Vieira, Rodolfo P; Lopes-Martins, Patrícia Sardinha Leonardo; Teixeira, Simone Aparecida; Borbely, Alexandre Urban; Gouvea, Irene Maria; Frigo, Lucio; Lopes-Martins, Rodrigo Álvaro Brandão

2015-01-01

221

Methyl p-Hydroxycinnamate Suppresses Lipopolysaccharide-Induced Inflammatory Responses through Akt Phosphorylation in RAW264.7 Cells  

PubMed Central

Derivatives of caffeic acid have been reported to possess diverse pharmacological properties such as anti-inflammatory, anti-tumor, and neuroprotective effects. However, the biological activity of methyl p-hydroxycinnamate, an ester derivative of caffeic acid, has not been clearly demonstrated. This study aimed to elucidate the anti-inflammatory mechanism of methyl p-hydroxycinnamate in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophage cells. Methyl p-hydroxycinnamate significantly inhibited LPS-induced excessive production of pro-inflammatory mediators such as nitric oxide (NO) and PGE2 and the protein expression of iNOS and COX-2. Methyl p-hydroxycinnamate also suppressed LPS-induced overproduction of pro-inflammatory cytokines such as IL-1? and TNF-?. In addition, methyl p-hydroxycinnamate significantly suppressed LPS-induced degradation of I?B, which retains NF-?B in the cytoplasm, consequently inhibiting the transcription of pro-inflammatory genes by NF-?B in the nucleus. Methyl p-hydroxycinnamate exhibited significantly increased Akt phosphorylation in a concentration-dependent manner. Furthermore, inhibition of Akt signaling pathway with wortmaninn abolished methyl p-hydroxycinnamate-induced Akt phosphorylation. Taken together, the present study clearly demonstrates that methyl p-hydroxycinnamate exhibits anti-inflammatory activity through the activation of Akt signaling pathway in LPS-stimulated RAW264.7 macrophage cells. PMID:24596616

Vo, Van Anh; Lee, Jae-Won; Shin, Seung-Yeon; Kwon, Jae-Hyun; Lee, Hee Jae; Kim, Sung-Soo; Kwon, Yong-Soo; Chun, Wanjoo

2014-01-01

222

The protective effect of alpha-lipoic Acid in lipopolysaccharide-induced acute lung injury is mediated by heme oxygenase-1.  

PubMed

Alpha-lipoic acid (ALA), occurring naturally in human food, is known to possess antioxidative and anti-inflammatory activities. Induction of heme oxygenase-1 (HO-1) has been reported to exhibit a therapeutic effect in several inflammatory diseases. The aim of study was to test the hypothesis that the protection of ALA against lipopolysaccharide-(LPS-) induced acute lung injury (ALI) is mediated by HO-1. Pre- or posttreatment with ALA significantly inhibited LPS-induced histological alterations of ALI, lung tissue edema, and production of proinflammatory cytokine, cytokine inducible neutrophil chemoattractant-3, and nitrite/nitrate in bronchoalveolar lavage fluid. In addition, the inflammatory responses including elevation of superoxide formation, myeloperoxidase activity, polymorphonuclear neutrophils infiltration, nitrotyrosine, inducible nitric oxide synthase expression and nuclear factor-kappa B (NF- ? B) activation in lung tissues of LPS-instilled rats were also markedly reduced by ALA. Interestingly, treatment with ALA significantly increased nuclear factor-erythroid 2-related factor 2 (Nrf2) activation and HO-1 expression in lungs of ALI. However, blocking HO-1 activity by tin protoporphyrin IX (SnPP), an HO-1 inhibitor, markedly abolished these beneficial effects of ALA in LPS-induced ALI. These results suggest that the protection mechanism of ALA may be through HO-1 induction and in turn suppressing NF- ? B-mediated inflammatory responses. PMID:23573137

Lin, Yu-Chieh; Lai, Yuan-Shu; Chou, Tz-Chong

2013-01-01

223

The Protective Effect of Alpha-Lipoic Acid in Lipopolysaccharide-Induced Acute Lung Injury Is Mediated by Heme Oxygenase-1  

PubMed Central

Alpha-lipoic acid (ALA), occurring naturally in human food, is known to possess antioxidative and anti-inflammatory activities. Induction of heme oxygenase-1 (HO-1) has been reported to exhibit a therapeutic effect in several inflammatory diseases. The aim of study was to test the hypothesis that the protection of ALA against lipopolysaccharide-(LPS-) induced acute lung injury (ALI) is mediated by HO-1. Pre- or posttreatment with ALA significantly inhibited LPS-induced histological alterations of ALI, lung tissue edema, and production of proinflammatory cytokine, cytokine inducible neutrophil chemoattractant-3, and nitrite/nitrate in bronchoalveolar lavage fluid. In addition, the inflammatory responses including elevation of superoxide formation, myeloperoxidase activity, polymorphonuclear neutrophils infiltration, nitrotyrosine, inducible nitric oxide synthase expression and nuclear factor-kappa B (NF-?B) activation in lung tissues of LPS-instilled rats were also markedly reduced by ALA. Interestingly, treatment with ALA significantly increased nuclear factor-erythroid 2-related factor 2 (Nrf2) activation and HO-1 expression in lungs of ALI. However, blocking HO-1 activity by tin protoporphyrin IX (SnPP), an HO-1 inhibitor, markedly abolished these beneficial effects of ALA in LPS-induced ALI. These results suggest that the protection mechanism of ALA may be through HO-1 induction and in turn suppressing NF-?B-mediated inflammatory responses. PMID:23573137

Lin, Yu-Chieh; Lai, Yuan-Shu; Chou, Tz-Chong

2013-01-01

224

Inducible Nitric Oxide Synthase Mediates Retinal Apoptosis in Ischemic Proliferative Retinopathy  

E-print Network

, 1994) and structural changes in a model of retinopathy of prematurity (Lachapelle et al., 1999 after the resolution of neovascular complications in retinopathy of prematurity (Fulton and Hansen, 1996Inducible Nitric Oxide Synthase Mediates Retinal Apoptosis in Ischemic Proliferative Retinopathy

Boyer, Edmond

225

Prostacyclin prevents nitric oxide-induced megakaryocyte apoptosis  

PubMed Central

We have previously demonstrated that nitric oxide (NO) triggers CD34+-derived megakaryocyte apoptosis. We here show that prostacyclin (PGI2) inhibits PAPA/NO-induced megakaryocyte death detected by fluorescent microscopy and flow cytometry. The cAMP-specific phosphodiesterase inhibitor, Ro 20-1724, and the permeable analog dibutyryl-cAMP also delayed apoptosis. PGI2 effect was fully prevented when adenylyl cyclase activity was suppressed by SQ 22536, and partially reversed by the permeable protein kinase A inhibitor PKI 14-22 amide. ELISA showed that while both PGI2 and NO alone or synergistically raised cAMP, only NO was able to increase intracellular cGMP levels. Treatment of megakaryocytes with PGI2 abolished both basal and NO-raised cGMP levels. Addition of 8-pCPT-cGMP or activation of soluble guanylyl cyclase by BAY 41-2272 induced cell death in a concentration-dependent manner, and ODQ, an inhibitor of guanylyl cyclase, prevented both PAPA/NO- or BAY 41-2272-induced apoptosis. Specific cGMP phosphodiesterase inhibition by Zaprinast or suppression of adenylyl cyclase by SQ 22536 enhanced the PAPA/NO proapoptotic effect. PGI2 completely inhibited NO-mediated generation and the increased activity of the cleaved form of caspase-3. In conclusion, our results demonstrate that contrary to their well-known direct and synergistic inhibitory effects on platelets, PGI2 and NO regulate opposite megakaryocyte survival responses through a delicate balance between intracellular cyclic nucleotide levels and caspase-3 activity control. PMID:15778737

Gabriel Pozner, Roberto; Negrotto, Soledad; D'Atri, Lina Paola; Lidia Kotler, Mónica; Angela Lazzari, María; Martin Gomez, Ricardo; Schattner, Mirta

2005-01-01

226

Inducible nitric oxide synthase of macrophages. Present knowledge and evidence for species-specific regulation  

Microsoft Academic Search

An important mechanism by which macrophages (M?) halt the growth of and eliminate a broad array of intracellular pathogens is the production of nitric oxide (NO). NO generation is catalyzed by inducible nitric oxide synthase (iNOS) converting arginine into citrulline and NO. In murine M?, iNOS activity is regulated largely at the transcriptional level. LPS and IFN-? induce iNOS, IL-4

T. W. Jungi; H. Adler; B. Adler; M. Thöny; M. Krampe; E. Peterhans

1996-01-01

227

In vivo Expression of Inducible Nitric Oxide Synthase in Experimentally Induced Neurologic Diseases  

NASA Astrophysics Data System (ADS)

The purpose of this study was to investigate the induction of inducible nitric oxide synthase (iNOS) mRNA in the brain tissue of rats and mice under the following experimental conditions: in rats infected with borna disease virus and rabies virus, in mice infected with herpes simplex virus, and in rats after the induction of experimental allergic encephalitis. The results showed that iNOS mRNA, normally nondetectable in the brain, was present in animals after viral infection or after induction of experimental allergic encephalitis. The induction of iNOS mRNA coincided with the severity of clinical signs and in some cases with the presence of inflammatory cells in the brain. The results indicate that nitric oxide produced by cells induced by iNOS may be the toxic factor accounting for cell damage and this may open the door to approaches to the study of the pathogenesis of neurological diseases.

Koprowski, Hilary; Zheng, Yong Mu; Heber-Katz, Ellen; Fraser, Nigel; Rorke, Lucy; Fu, Zhen Fang; Hanlon, Cathleen; Dietzschold, Bernhard

1993-04-01

228

Somatostatin prevents lipopolysaccharide?induced neurodegeneration in the rat substantia nigra by inhibiting the activation of microglia.  

PubMed

Somatostatin (SST) is a neuromodulator which is abundant throughout the central nervous system (CNS) and has a crucial role in neurodegenerative disorders. However, little is known about the effects and mechanisms of SST in dopaminergic (DA) neurons in the context of Parkinson's disease (PD). In the present study, a model of PD was generated by injecting lipopolysaccharide (LPS) into the substantia nigra (SN) of rats in order to investigate the effects of SST on LPS?induced degeneration of DA in vivo. Intramural injection of LPS resulted in a significant loss of DA neurons, while reduction of neuronal death by SST pretreatment was confirmed using immunohistochemical staining for tyrosine hydroxylase and Nissl. In parallel, immunohistochemical detection of OX?42 and hydroethidine staining were employed to determine the activation of microglia and production of reactive oxygen species (ROS), respectively. It was found that SST inhibited the LPS?induced microglial activity and ROS production. ELISA revealed a decreased production of pro?inflammatory mediators, including tumor necrosis factor??, interleukin?1? and prostaglandin E2 when SST was administered prior to LPS treatment. Western blot analysis showed that LPS?induced expression of inducible nitric oxide synthase, cyclooxygenase?2 and nuclear factor ?B (NF??B) p?p65 was attenuated by administration of SST prior to LPS application. The results indicated that LPS?induced loss of nigral DA neurons was inhibited by SST and the observed effects of SST on neuroprotection were associated with suppression of microglial activation and the NF??B pathway, ensuing decreases of neuroinflammation and oxidative stress. The present study therefore suggested that SST is beneficial for treating neurodegenerative diseases, such as PD, through inhibiting the activation of microglia. PMID:25777539

Bai, Lijuan; Zhang, Xique; Li, Xiaohong; Liu, Na; Lou, Fan; Ma, Honglei; Luo, Xiaoguang; Ren, Yan

2015-07-01

229

Phosphorylation of Akt Mediates Anti-Inflammatory Activity of 1-p-Coumaroyl ?-D-Glucoside Against Lipopolysaccharide-Induced Inflammation in RAW264.7 Cells  

PubMed Central

Hydroxycinnamic acids have been reported to possess numerous pharmacological activities such as antioxidant, anti-inflammatory, and anti-tumor properties. However, the biological activity of 1-p-coumaroyl ?-D-glucoside (CG), a glucose ester derivative of p-coumaric acid, has not been clearly examined. The objective of this study is to elucidate the anti-inflammatory action of CG in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells. In the present study, CG significantly suppressed LPS-induced excessive production of pro-inflammatory mediators such as nitric oxide (NO) and PGE2 and the protein expression of iNOS and COX-2. CG also inhibited LPS-induced secretion of pro-inflammatory cytokines, IL-1? and TNF-?. In addition, CG significantly suppressed LPS-induced degradation of I?B. To elucidate the underlying mechanism by which CG exerts its anti-inflammatory action, involvement of various signaling pathways were examined. CG exhibited significantly increased Akt phosphorylation in a concentration-dependent manner, although MAPKs such as Erk, JNK, and p38 appeared not to be involved. Furthermore, inhibition of Akt/PI3K signaling pathway with wortmannin significantly, albeit not completely, abolished CG-induced Akt phosphorylation and anti-inflammatory actions. Taken together, the present study demonstrates that Akt signaling pathway might play a major role in CG-mediated anti-inflammatory activity in LPS-stimulated RAW264.7 macrophage cells. PMID:24634601

Vo, Van Anh; Lee, Jae-Won; Kim, Ji-Young; Park, Jun-Ho; Lee, Hee Jae; Kim, Sung-Soo; Kwon, Yong-Soo

2014-01-01

230

Regulation of Inducible Nitric Oxide Synthase Gene in Glial Cells  

PubMed Central

Elevated levels of NO produced within the central nervous system (CNS) are associated with the pathogenesis of neuroinflammatory and neurodegenerative human diseases such as multiple sclerosis, HIV dementia, brain ischemia, trauma, Parkinson's disease, and Alzheimer's disease. Resident glial cells in the CNS (astroglia and microglia) express inducible nitric oxide synthase (iNOS) and produce high levels of NO in response to a wide variety of proinflammatory and degenerative stimuli. Although pathways resulting in the expression of iNOS may vary in two different glial cells of different species, the intracellular signaling events required for the expression of iNOS in these cells are slowly becoming clear. Various signaling cascades converge to activate several transcription factors that control the transcription of iNOS in glial cells. The present review summarizes different results and discusses current understandings about signaling mechanisms for the induction of iNOS expression in activated glial cells. A complete understanding of the regulation of iNOS expression in glial cells is expected to identify novel targets for therapeutic intervention in NO-mediated neurological disorders. PMID:16771683

Saha, Ramendra N.; Pahan, Kalipada

2007-01-01

231

Aldose reductase regulates TNF-?-induced inducible nitric oxide synthase expression in human mesangial cells.  

PubMed

Glomerulonephritis is one of the most important causes of renal failure, which is accompanied with production of Nitric Oxide (NO) synthesized by inducible nitric oxide synthase (iNOS). Aldose reductase (AR) is the key enzyme in polyol pathway and plays an important role in glucose metabolism. Here, we report our finding that AR regulates tumor-necrosis-factor-?-induced (TNF-?-induced) iNOS expression in human mesangial cells (HMC) via nuclear factor ?B (NF?B) signal pathway. The TNF-?-induced iNOS expression in HMC with different level of AR was measured by Real-time PCR and Western blot. The activation of signal pathway was analyzed by Western blot and electrophoretic mobility shift assay (EMSA). In cultured HMC, TNF-? induces the expression of iNOS, which is attenuated by knockdown AR with siRNA. On the other side, transfection with pcDNA3.1-AR gets the consistent conclusion. Furthermore, knockdown of AR attenuated activity of NF?B, suggesting that the effects of AR on this pathway may result in the reduced iNOS transcription and expression. Taken together, these data demonstrate the role of AR in regulating iNOS expression induced by TNF-? in cultured HMC, indicating the novel function of AR in glomerulonephritis besides glucose metabolism. PMID:21637955

Zhao, Jingjing; Jiang, Tao; Li, Hui; Zhang, Yuejuan; Zhang, Nong

2012-02-01

232

Modified peptide nucleic acids are internalized in mouse macrophages RAW 264.7 and inhibit inducible nitric oxide synthase  

Microsoft Academic Search

Overexpression of inducible nitric oxide synthase causes the production of high levels of nitric oxide, which, under pathological conditions, leads to immunosuppression and tissue damage. The results recently obtained using peptide nucleic acids, rather than traditional oligonucleotides as antigen and antisense molecules, prompted us to test their efficacy in the regulation of nitric oxide production, thereby overcoming the obstacle of

Sonia Scarfi; Marco Giovine; Anna Gasparini; Gianluca Damonte; Enrico Millo; Marina Pozzolini; Umberto Benatti

1999-01-01

233

Nilotinib ameliorates lipopolysaccharide-induced acute lung injury in rats  

SciTech Connect

The present study aimed to investigate the effect of the new tyrosine kinase inhibitor, nilotinib on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in rats and explore its possible mechanisms. Male Sprague-Dawley rats were given nilotinib (10 mg/kg) by oral gavage twice daily for 1 week prior to exposure to aerosolized LPS. At 24 h after LPS exposure, bronchoalveolar lavage fluid (BALF) samples and lung tissue were collected. The lung wet/dry weight (W/D) ratio, protein level and the number of inflammatory cells in the BALF were determined. Optical microscopy was performed to examine the pathological changes in lungs. Malondialdehyde (MDA) content, superoxidase dismutase (SOD) and reduced glutathione (GSH) activities as well as nitrite/nitrate (NO{sub 2}{sup -}/NO{sub 3}{sup -}) levels were measured in lung tissues. The expression of inflammatory cytokines, tumor necrosis factor-{alpha} (TNF-{alpha}), transforming growth factor-{beta}{sub 1} (TGF-{beta}{sub 1}) and inducible nitric oxide synthase (iNOS) were determined in lung tissues. Treatment with nilotinib prior to LPS exposure significantly attenuated the LPS-induced pulmonary edema, as it significantly decreased lung W/D ratio, protein concentration and the accumulation of the inflammatory cells in the BALF. This was supported by the histopathological examination which revealed marked attenuation of LPS-induced ALI in nilotinib treated rats. In addition, nilotinib significantly increased SOD and GSH activities with significant decrease in MDA content in the lung. Nilotinib also reduced LPS mediated overproduction of pulmonary NO{sub 2}{sup -}/NO{sub 3}{sup -} levels. Importantly, nilotinib caused down-regulation of the inflammatory cytokines TNF-{alpha}, TGF-{beta}{sub 1} and iNOS levels in the lung. Taken together, these results demonstrate the protective effects of nilotinib against the LPS-induced ALI. This effect can be attributed to nilotinib ability to counteract the inflammatory cells infiltration and hence ROS generation and regulate cytokine effects. - Research highlights: > The protective effects of nilotinib against LPS-induced ALI in rats were studied. > Nilotinib showed potent anti-inflammatory activity as it attenuated PMN infiltration and hence ROS generation. > In addition, nilotinib caused down-regulation of proinflammatory cytokine production.

El-Agamy, Dina S., E-mail: dinaagamy1@yahoo.com

2011-06-01

234

Isobavachalcone suppresses expression of inducible nitric oxide synthase induced by Toll-like receptor agonists.  

PubMed

Toll-like receptors (TLRs) play an important role by recognizing many pathogen-associated molecular patterns and inducing innate immunity. Dysregulated activation of TLR signaling pathways induces the activation of various transcription factors such as nuclear factor-?B, leading to the induction of pro-inflammatory gene products such as inducible nitric oxide synthase (iNOS). The present study investigated the effect of isobavachalcone (IBC), a natural chalcone component of Angelica keiskei, on inflammation by modulating iNOS expression induced by TLR agonists in murine macrophages. IBC suppressed iNOS expression induced by macrophage-activating lipopeptide 2-kDa, polyriboinosinic polyribocytidylic acid, or lipopolysaccharide. These results indicate the potential of IBC as a potent anti-inflammatory drug. PMID:23164691

Shin, Hwa-Jeong; Shon, Dong-Hwa; Youn, Hyung-Sun

2013-01-01

235

Dissecting structural and electronic effects in inducible nitric oxide synthase.  

PubMed

Nitric oxide synthases (NOSs) are haem-thiolate enzymes that catalyse the conversion of L-arginine (L-Arg) into NO and citrulline. Inducible NOS (iNOS) is responsible for delivery of NO in response to stressors during inflammation. The catalytic performance of iNOS is proposed to rely mainly on the haem midpoint potential and the ability of the substrate L-Arg to provide a hydrogen bond for oxygen activation (O-O scission). We present a study of native iNOS compared with iNOS-mesohaem, and investigate the formation of a low-spin ferric haem-aquo or -hydroxo species (P) in iNOS mutant W188H substituted with mesohaem. iNOS-mesohaem and W188H-mesohaem were stable and dimeric, and presented substrate-binding affinities comparable to those of their native counterparts. Single turnover reactions catalysed by iNOSoxy with L-Arg (first reaction step) or N-hydroxy-L-arginine (second reaction step) showed that mesohaem substitution triggered higher rates of FeIIO2 conversion and altered other key kinetic parameters. We elucidated the first crystal structure of a NOS substituted with mesohaem and found essentially identical features compared with the structure of iNOS carrying native haem. This facilitated the dissection of structural and electronic effects. Mesohaem substitution substantially reduced the build-up of species P in W188H iNOS during catalysis, thus increasing its proficiency towards NO synthesis. The marked structural similarities of iNOSoxy containing native haem or mesohaem indicate that the kinetic behaviour observed in mesohaem-substituted iNOS is most heavily influenced by electronic effects rather than structural alterations. PMID:25608846

Hannibal, Luciana; Page, Richard C; Haque, Mohammad Mahfuzul; Bolisetty, Karthik; Yu, Zhihao; Misra, Saurav; Stuehr, Dennis J

2015-04-01

236

Garlic prevents hypertension induced by chronic inhibition of nitric oxide synthesis  

Microsoft Academic Search

It has been reported that garlic activates nitric oxide synthase in vitro and that chronic inhibition of nitric oxide (NO) synthesis by N?-nitro-L-arginine-methyl-ester (L-NAME) induces arterial hypertension in rats. In this work, we studied the effect of oral administration of L-NAME for 4 weeks on control and garlic-fed rats. Basal systolic blood pressure was recorded 4 weeks after garlic supplementation,

José Pedraza-Chaverrí; Edilia Tapia; Omar N. Medina-Campos; Martha Franco

1998-01-01

237

Energetic particle-induced enhancements of stratospheric nitric acid  

NASA Technical Reports Server (NTRS)

Inclusion of complete ion chemistry in the calculation of minor species production during energetic particle deposition events leads to significant enhancement in the calculated nitric acid concentration during precipitation. An ionization rate of 1.2 x 10(exp 3)/cu cm/s imposed for 1 day increases HNO3 from 3 x 10(exp 5) to 6 x 10(exp 7)/cu cm at 50 km. With an ionization rate of 600 cu cm/s, the maximum HNO3 is 3 x 10(exp 7)/cu cm. Calculations which neglect negative ions predict the nitric acid will fall during precipitation events. The decay time for converting HNO3 into odd nitrogen and hydrogen is more than 1 day for equinoctial periods at 70 deg latitude. Examination of nitric acid data should yield important information on the magnitude and frequency of charged particle events.

Aikin, Arthur C.

1994-01-01

238

Spirulina Promotes Stem Cell Genesis and Protects against LPS Induced Declines in Neural Stem Cell Proliferation  

Microsoft Academic Search

Adult stem cells are present in many tissues including, skin, muscle, adipose, bone marrow, and in the brain. Neuroinflammation has been shown to be a potent negative regulator of stem cell and progenitor cell proliferation in the neurogenic regions of the brain. Recently we demonstrated that decreasing a key neuroinflammatory cytokine IL-1? in the hippocampus of aged rats reversed the

Adam D. Bachstetter; Jennifer Jernberg; Andrea Schlunk; Jennifer L. Vila; Charles Hudson; Michael J. Cole; R. Douglas Shytle; Jun Tan; Paul R. Sanberg; Cyndy D. Sanberg; Cesario Borlongan; Yuji Kaneko; Naoki Tajiri; Carmelina Gemma; Paula C. Bickford; Tsuneya Ikezu

2010-01-01

239

Social management of LPS-induced inflammation in Formica polyctena ants  

Microsoft Academic Search

Invertebrates, and especially insects, constitute valuable and convenient models for the study of the evolutionary roots of immune-related behaviors. With stable conditions in the nest, high population densities, and frequent interactions, social insects such as ants provide an excellent system for examining the spread of pathogens. The evolutionary success of these species raises questions about the behavioral responses of social

A. Aubert; F.-J. Richard

2008-01-01

240

Regulation of LPS-induced tissue factor expression in human monocytic THP-1 cells by curcumin  

Technology Transfer Automated Retrieval System (TEKTRAN)

Tissue factor (TF) is a transmembrane receptor, which initiates thrombotic episodes associated with various diseases. In addition to membrane-bound TF, we have discovered an alternatively spliced form of human TF mRNA. It was later confirmed that this form of TF mRNA expresses a soluble protein circ...

241

Previous Burn Injury Predisposes Mice to Lipopolysaccharide (LPS) Induced Changes in Glucose Metabolism  

PubMed Central

In mice, it has been demonstrated that at 7 days after burn injury, injection of LPS is more lethal than the same dose at one day after injury. In the present study, we examined the effect of LPS injection to mice burned seven days previously on glucose metabolism (18FDG uptake) in vivo. CD-1 male mice (25-28 grams, Charles River breeding laboratories) were anesthetized, backs shaven, and subjected to dorsal full thickness burn on 25% total body surface area. Sham treated animals were used as controls. Six days after burn injury all mice were fasted overnight. One half of the burned and sham controls were subsequently injected i.p. with LPS (10 mg/kg, E. Coli). The remaining animals were injected with saline i.p. Two hours later all mice were injected i.v. with 50 ?Ci of 18F FDG. One hour later the animals were euthanized and biodistribution was measured. Tissues were weighed and radioactivity was measured with a well-type gamma counter. Results were expressed as %dose/gram tissue, mean ± SEM. The combination of burn 7 days previously and LPS significantly increased mortality compared animals with burn alone, LPS alone or sham controls. Burn injury seven days previously caused a significant reduction in 18FDG uptake by the brain compared to sham controls. The combination of LPS and burn injury seven days previously produced a significant increase in 18FDG uptake by brown adipose tissue (BAT) and heart compared with either treatment separately. LPS produced a significant increase in 18FDG uptake by lung, spleen and GI tract of the sham animals, changes that were different in mice burned 7 days previously and injected with LPS. The present results suggest that burn injury seven days previously predisposes mice to alterations in 18FDG uptake produced by LPS. These changes may relate in part, to the increased lethality of LPS injection in previously burned mice. PMID:22961012

Carter, Edward A.; Paul, Kasie; Barrow, Sandra A.; Fischman, Alan J.; Tompkins, Ronald G.

2012-01-01

242

LPS-Induced Genes in Intestinal Tissue of the Sea Cucumber Holothuria glaberrima  

PubMed Central

Metazoan immunity is mainly associated with specialized cells that are directly involved with the immune response. Nevertheless, both in vertebrates and invertebrates other organs might respond to immune activation and participate either directly or indirectly in the ongoing immune process. However, most of what is known about invertebrate immunity has been restricted to immune effector cells and little information is available on the immune responses of other tissues or organs. We now focus on the immune reactions of the intestinal tissue of an echinoderm. Our study employs a non-conventional model, the echinoderm Holothuria glaberrima, to identify intestinal molecules expressed after an immune challenge presented by an intra-coelomic injection of lipopolysaccharides (LPS). The expression profiles of intestinal genes expressed differentially between LPS-injected animals and control sea water-injected animals were determined using a custom-made Agilent microarray with 7209 sea cucumber intestinal ESTs. Fifty (50) unique sequences were found to be differentially expressed in the intestine of LPS-treated sea cucumbers. Seven (7) of these sequences represented homologues of known proteins, while the remaining (43) had no significant similarity with any protein, EST or RNA database. The known sequences corresponded to cytoskeletal proteins (Actin and alpha-actinin), metabolic enzymes (GAPDH, Ahcy and Gnmt), metal ion transport/metabolism (major yolk protein) and defense/recognition (fibrinogen-like protein). The expression pattern of 11 genes was validated using semi-quantitative RT-PCR. Nine of these corroborated the microarray results and the remaining two showed a similar trend but without statistical significance. Our results show some of the molecular events by which the holothurian intestine responds to an immune challenge and provide important information to the study of the evolution of the immune response. PMID:19584914

Ramírez-Gómez, Francisco; Ortiz-Pineda, Pablo A.; Rivera-Cardona, Gabriela; García-Arrarás, José E.

2009-01-01

243

AURANOFIN, AS AN ANTI-RHEUMATIC GOLD COMPOUND SUPPRESSES LPS-INDUCED HOMODIMERIZATION OF TLR4  

Technology Transfer Automated Retrieval System (TEKTRAN)

Toll-like receptors (TLRs), which are activated by invading microorganisms or endogenous molecules, evoke immune and inflammatory responses. TLR activation is closely linked to the development of many chronic inflammatory diseases including rheumatoid arthritis. Auranofin, an Au(I) compound, is a we...

244

Rationally Designed Macrocyclic Peptides as Synergistic Agonists of LPS-Induced Inflammatory Response.  

PubMed

Toll-like receptor 4 (TLR4) plays an important role in the regulation of the innate and adaptive immune response. Both agonists and antagonists of TLR4 are of considerable interest as drug leads for various disease indications. We herein report the rational design of two myeloid differentiation factor 2 (MD2)-derived macrocyclic peptides as TLR4 modulators, using the Rosetta Macromolecular Modeling software. The designed cyclic peptides, but not their linear counterparts, displayed synergistic activation of TLR signaling when co-administered with lipopolysaccharide (LPS). Although the understanding of the mechanism of action of these peptides remains elusive; these results underscore the utility of peptide cyclization for the discovery of biologically active agents, and also lead to valuable tools for the investigation of TLR4 signaling. PMID:25400297

Gao, Meng; London, Nir; Cheng, Kui; Tamura, Ryo; Jin, Jialin; Schueler-Furman, Ora; Yin, Hang

2014-10-21

245

Transcriptional profiling of the LPS induced NF-?B response in macrophages  

Microsoft Academic Search

BACKGROUND: Exposure of macrophages to bacterial products such as lipopolysaccharide (LPS) results in activation of the NF-?B transcription factor, which orchestrates a gene expression programme that underpins the macrophage-dependent immune response. These changes include the induction or repression of a wide range of genes that regulate inflammation, cell proliferation, migration and cell survival. This process is tightly regulated and loss

Omar Sharif; Viacheslav N Bolshakov; Stephanie Raines; Peter Newham; Neil D Perkins

2007-01-01

246

LPS-induced chorioamnionitis and antenatal corticosteroids modulate Shh signaling in the ovine fetal lung  

PubMed Central

Chorioamnionitis and antenatal corticosteroids mature the fetal lung functionally but disrupt late-gestation lung development. Because Sonic Hedgehog (Shh) signaling is a major pathway directing lung development, we hypothesized that chorioamnionitis and antenatal corticosteroids modulated Shh signaling, resulting in an altered fetal lung structure. Time-mated ewes with singleton ovine fetuses received an intra-amniotic injection of lipopolysaccharide (LPS) and/or maternal intramuscular betamethasone 7 and/or 14 days before delivery at 120 days gestational age (GA) (term = 150 days GA). Intra-amniotic LPS exposure decreased Shh mRNA levels and Gli1 protein expression, which was counteracted by both betamethasone pre- or posttreatment. mRNA and protein levels of fibroblast growth factor 10 and bone morphogenetic protein 4, which are important mediators of lung development, increased 2-fold and 3.5-fold, respectively, 14 days after LPS exposure. Both 7-day and 14-day exposure to LPS changed the mRNA levels of elastin (ELN) and collagen type I alpha 1 (Col1A1) and 2 (Col1A2), which resulted in fewer elastin foci and increased collagen type I deposition in the alveolar septa. Corticosteroid posttreatment prevented the decrease in ELN mRNA and increased elastin foci and decreased collagen type I deposition in the fetal lung. In conclusion, fetal lung exposure to LPS was accompanied by changes in key modulators of lung development resulting in abnormal lung structure. Betamethasone treatment partially prevented the changes in developmental processes and lung structure. This study provides new insights into clinically relevant prenatal exposures and fetal lung development. PMID:22962010

Collins, Jennifer J. P.; Kuypers, Elke; Nitsos, Ilias; Jane Pillow, J.; Polglase, Graeme R.; Kemp, Matthew W.; Newnham, John P.; Cleutjens, Jack P.; Frints, Suzanna G. M.; Kallapur, Suhas G.; Jobe, Alan H.

2012-01-01

247

LPS-Induced Galectin-3 Oligomerization Results in Enhancement of Neutrophil Activation  

Microsoft Academic Search

Galectin-3 (Gal 3) is a glycan-binding protein that can be secreted by activated macrophages and mast cells at inflammation sites and plays an important role in inflammatory diseases caused by Bacteria and their products, such as lipopolysaccharides (LPS). Although it is well established that Gal 3 can interact with LPS, the pathophysiological importance of LPS\\/Gal 3 interactions is not fully

Marise Lopes Fermino; Claudia Danella Polli; Karina Alves Toledo; Fu-Tong Liu; Dan K. Hsu; Maria Cristina Roque-Barreira; Gabriela Pereira-da-Silva; Emerson Soares Bernardes; Lise Halbwachs-Mecarelli; Jean-Pierre Gorvel

2011-01-01

248

ROLE OF CELL SIGNALING IN PROTECTION FROM DIESEL AND LPS INDUCED ACUTE LUNG INJURY  

EPA Science Inventory

We have previously demonstrated in CD-1 mice that pre-administration of N-acetyl cysteine (NAC) or the p38 MAP kinase inhibitor (SB203580) reduces acute lung injury and inflammation following pulmonary exposures to diesel exhaust particles (DEP) or lipopolysaccharide (LPS). Here ...

249

AFFYMETRIX GENECHIP-BASED ANALYSIS OF THE GENOMIC RESPONSE TO ACUTE LPS-INDUCED BOVINE MASTITIS  

Technology Transfer Automated Retrieval System (TEKTRAN)

The genomic response of the bovine mammary gland was profiled four hours after an intramammary challenge with E. coli endotoxin (LPS). Three mid-lactation cows were challenged in one quarter with 1 ug of LPS while contralateral quarters received saline and served as within animal controls. RNA from ...

250

Roquefort Cheese Proteins Inhibit Chlamydia pneumoniae Propagation and LPS-Induced Leukocyte Migration  

PubMed Central

Inflammation in atherosclerosis, which could be associated with some subclinical infections such as C. pneumoniae, is one of the key factors responsible for the development of clinical complications of this disease. We report that a proprietary protein extract isolated from Roquefort cheese inhibits the propagation of C. pneumoniae in a human HL cell line in a dose-dependent manner, as revealed by the immunofluorescence analysis. These changes were accompanied by a significant reduction in the infective progeny formation over the protein extract range of 0.12–0.5??g/mL. Moreover, short term feeding of mice with Roquefort cheese (twice, 10?mg per mouse with an interval of 24 hours) led to the inhibition of the migration of peritoneal leukocytes caused by intraperitoneal injection of E. coli lipopolysaccharide. These changes were complemented by a reduction in neutrophil count and a relative increase in peritoneal macrophages, suggesting that ingestion of Roquefort could promote regenerative processes at the site of inflammation. The ability of this protein to inhibit propagation of Chlamydia infection, as well as the anti-inflammatory and proregenerative effects of Roquefort itself, may contribute to the low prevalence of cardiovascular mortality in France where consumption of fungal fermented cheeses is the highest in the world. PMID:23737705

Petyaev, Ivan M.; Zigangirova, Naylia A.; Kobets, Natalie V.; Tsibezov, Valery; Kapotina, Lydia N.; Fedina, Elena D.; Bashmakov, Yuriy K.

2013-01-01

251

Isoflavone-free soy protein diet inhibits LPS-induced inflammatory responses  

Technology Transfer Automated Retrieval System (TEKTRAN)

Recently, we showed reduced atherosclerotic lesions in a hyperlipidemic mouse model fed isoflavone-free soy protein diet (SPI–) compared to casein (CAS)-fed mice, despite unchanged serum lipid levels. However, the molecular mechanisms contributing to the atheroprotective effect of soy-based diets is...

252

The chloroform fraction of Solanum nigrum suppresses nitric oxide and tumor necrosis factor-? in LPS-stimulated mouse peritoneal macrophages through inhibition of p38, JNK and ERK1/2.  

PubMed

Solanum nigrum L., commonly known as black nightshade, is used worldwide for the treatment of skin and mucosal ulcers, liver cirrhosis and edema. We aimed to determine the anti-inflammatory active fraction of S. nigrum by serial extractions. S. nigrum was first extracted with methanol, then fractionated with chloroform and water. The effects of S. nigrum fractions, diosgenin and ?-solanine on LPS/interferon-gamma-induced nitric oxide (NO) and inducible NO synthase (iNOS), or LPS-induced tumor necrosis factor-? (TNF-?) and interleukin (IL)-6, in mouse peritoneal macrophages were determined. Western blotting analysis was used to detect LPS-induced phosphorylation of p38, JNK and ERK1/2. The chloroform fraction of S. nigrum was cytotoxic in a time and concentration dependent manner; however, the methanol and water fractions were not. The chloroform fraction reduced NO through inhibition of iNOS synthesis and inhibited TNF-? and IL-6 at the level of protein secretion; the methanol and water fractions showed a weak or no effect. The chloroform fraction also suppressed p38, JNK and ERK1/2. Diosgenin and ?-solanine were cytotoxic at a high concentration. In particular, diosgenin was able to inhibit TNF-? and IL-6, but both compounds did not affect LPS-induced iNOS expression. These results indicate that the anti-inflammatory compounds of S. nigrum exist preferentially in the nonpolar fraction, ruling out the possibility that diosgenin and ?-solanine are the likely candidates. The inhibition of iNOS, TNF-? and IL-6 by the chloroform fraction may be partly due to the suppression of p38, JNK and ERK1/2. Further study is required to identify the active compounds of S. nigrum. PMID:22083995

Kang, Hee; Jeong, Ha-Deok; Choi, Ho-Young

2011-01-01

253

Nitrate Reductase is Responsible for Elicitin-induced Nitric Oxide Production in Nicotiana benthamiana  

Microsoft Academic Search

; Recent works have established a key role for nitric oxide (NO) in activating disease resistance in plants. Nitrate reductase (NR) is one of the enzymes that are capable of producing NO in plants. In a previous study, we reported that pathogen signals induce expression of NR genes in potato, suggesting the involvement of NR in NO produc- tion induced

Ayako Yamamoto-Katou; Shinpei Katou; Hirofumi Yoshioka; Noriyuki Doke; Kazuhito Kawakita

2006-01-01

254

Role of inducible nitric oxide synthase expression and peroxynitrite formation in guinea pig ileitis  

Microsoft Academic Search

Background & Aims Inflammatory bowel disease is characterized by increased synthesis of nitric oxide. The aim of this study was to determine if inducible NO synthase (iNOS) was responsible for tissue injury, potentially via peroxynitrite formation, in the guinea pig model of gut inflammation. Methods Inflammation was induced in guinea pig ileum by intraluminal administration of the hapten trinitrobenzene sulfonic

Mark J. S. Miller; Jane H. Thompson; Xiao-Jing Zhang; Halina Sadowska-Krowicka; Jane L. Kakkis; Upender K. Munshi; Manuel Sandoval; Janet L. Rossi; Sandra Eloby-Childress; Joseph S. Beckman; Yao Zu Ye; Charles P. Rodi; Pamela T. Manning; Mark G. Currie; David A. Clark

1995-01-01

255

Cyclooxygenase-2-derived prostacyclin protective role on endotoxin-induced mouse cardiomyocyte mortality.  

PubMed

Cardiovascular dysfunction characterizes septic shock, inducing multiple organ failure and a high mortality rate. In the heart, it has been shown an up-regulation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expressions with subsequent overproduction of nitric oxide (NO) and eicosanoids. This study is focused on the links between these products of inflammation and cell loss of mouse cardiomyocytes during treatment by the Salmonella typhimurium lipopolysaccharide (LPS) in presence or in absence of NOS or COX inhibitors. LPS induced RelA/NF-?B p65 activation, iNOS and COX-2 up-regulations, resulting in NO and prostacyclin releases. These effects were reversed by the NO-synthase inhibitor and increased by the specific COX-2 inhibitor. Immunostainings with FITC-conjugated anti-Annexin-V and propidium iodide and caspase 3/7 activity assay showed that cardiomyocyte necrosis was inhibited by L-NA during LPS treatment challenge, while apoptosis was induced in presence of both LPS and NS-398. No effect on LPS cellular injury was observed using the specific cyclooxygenase-1 (COX-1) inhibitor, SC-560. These findings strongly support the hypothesis of a link between iNOS-dependent NO overproduction and LPS-induced cell loss with a selective protective role allotted to COX-2 and deriving prostacyclins. PMID:21769544

Panaro, Maria Antonietta; Pricci, Maria; Meziani, Ferhat; Ragot, Thierry; Andriantsitohaina, Ramaroson; Mitolo, Vincenzo; Tesse, Angela

2011-12-01

256

Inhibitory effect of sulphated polysaccharide porphyran on nitric oxide production in lipopolysaccharide-stimulated RAW264.7 macrophages.  

PubMed

Porphyran, extracted from an edible red alga (Porphyra yezoensis), is a sulphated polysaccharide with a wide variety of biological activities including anti-tumour, antioxidant and immuno-modulating activities. In this study, we examined the effect of porphyran on nitric oxide (NO) production in mouse macrophage cell line RAW264.7 cells. Although no significant activity of porphyran to induce NO or tumour necrosis factor-? (TNF-?) production in RAW264.7 cells was observed at the concentration range tested (10-500 µg/ml), it was found for the first time that porphyran inhibited NO production and expression of inducible nitric oxide synthase (iNOS) in RAW264.7 cells stimulated with lipopolysaccharide (LPS). In the presence of 500 µg/ml porphyran, NO production and expression of iNOS in LPS-treated RAW264.7 cells were completely suppressed. On the other hand, porphyran showed only a marginal effect on the secretion of TNF-? from LPS-stimulated RAW264.7 cells. Electrophoretic mobility shift assay (EMSA) using infrared dye labelled oligonucleotide with nuclear factor-?B (NF-?B) consensus sequence suggested that porphyran inhibited the LPS-induced NF-?B activation. The LPS-inducible nuclear translocation of p65, and the phosphorylation and degradation of I?B-? were also inhibited by the pre-treatment with porphyran. Our results obtained in in vitro analysis suggest that porphyran suppresses NO production in LPS-stimulated macrophages by the blocking of NF-?B activation. PMID:21976706

Jiang, Zedong; Hama, Yoichiro; Yamaguchi, Kenichi; Oda, Tatsuya

2012-01-01

257

The triterpenoid quinonemethide pristimerin inhibits induction of inducible nitric oxide synthase in murine macrophages  

Microsoft Academic Search

Inducible nitric oxide synthase dependent production of nitric oxide (NO) plays an important role in inflammation. We investigated whether pristimerin ((20?)-3-hydroxy-2-oxo-24-nor-friedela-1(10),3,5,7-tetraen-carboxylic acid-(29)-methylester), an antitumoral, antimicrobial as well as anti-inflammatory plant compound, has an effect on the inducible NO synthase system in lipopolysaccharide-activated RAW 264.7 macrophages. Pristimerin dose dependently (IC50: 0.2–0.3 ?M) reduces nitrite accumulation, a parameter for NO synthesis, in

Verena M Dirsch; Alexandra K Kiemer; Hildebert Wagner; Angelika M Vollmar

1997-01-01

258

Intermittent fasting attenuates lipopolysaccharide-induced neuroinflammation and memory impairment  

PubMed Central

Background Systemic bacterial infections often result in enduring cognitive impairment and are a risk factor for dementia. There are currently no effective treatments for infection-induced cognitive impairment. Previous studies have shown that intermittent fasting (IF) can increase the resistance of neurons to injury and disease by stimulating adaptive cellular stress responses. However, the impact of IF on the cognitive sequelae of systemic and brain inflammation is unknown. Methods Rats on IF for 30 days received 1 mg/kg of lipopolysaccharide (LPS) or saline intravenously. Half of the rats were subjected to behavioral tests and the other half were euthanized two hours after LPS administration and the hippocampus was dissected and frozen for analyses. Results Here, we report that IF ameliorates cognitive deficits in a rat model of sepsis by a mechanism involving NF-?B activation, suppression of the expression of pro-inflammatory cytokines, and enhancement of neurotrophic support. Treatment of rats with LPS resulted in deficits in cognitive performance in the Barnes maze and inhibitory avoidance tests, without changing locomotor activity, that were ameliorated in rats that had been maintained on the IF diet. IF also resulted in reduced levels of mRNAs encoding the LPS receptor TLR4 and inducible nitric oxide synthase (iNOS) in the hippocampus. Moreover, IF prevented LPS-induced elevation of IL-1?, IL-1? and TNF-? levels, and prevented the LPS-induced reduction of BDNF levels in the hippocampus. IF also significantly attenuated LPS-induced elevations of serum IL-1?, IFN-?, RANTES, TNF-? and IL-6 levels. Conclusions Taken together, our results suggest that IF induces adaptive responses in the brain and periphery that can suppress inflammation and preserve cognitive function in an animal model of systemic bacterial infection. PMID:24886300

2014-01-01

259

Isolation of benzoic and cinnamic acid derivatives from the grains of Sorghum bicolor and their inhibition of lipopolysaccharide-induced nitric oxide production in RAW 264.7 cells.  

PubMed

Two new caffeoylglycerols, (hwanggeumchal A-B, 9-10), together with eight known derivatives (1-8) were isolated from the grains of Sorghum bicolor (L.) Moench var. hwanggeumchal. Their chemical structures were established mainly by 1D and 2D NMR techniques and MS data analysis. Amongst those, methyl 3,4-dihydroxybenzoate (2), 3,4,5-trihydroxycinnamate (3), methyl 3,4-dihydroxycinnamate (5), caffeoylglycolic acid methyl ester (7) and 1-O-caffeoylglycerol (8) displayed potential inhibitory effects against LPS-induced NO production in macrophage RAW264.7 cells with IC50 values of 11.9, 2.9, 27.1, 29.0 and 18.5?M, respectively. Furthermore, these compounds dose-dependently reduced the LPS-induced iNOS expression. In addition, pre-incubation of cells with compounds 2, 3 and 5 significantly suppressed LPS-induced COX-2 protein expression. SAR investigation revealed that compounds with a methyl ester (COOCH3) group possessed stronger activity. Our obtained data suggest that S. bicolor and its caffeoylglyceride-enrich extracts may be applied as supplemental and/or functional foods having a beneficial effect against inflammation. PMID:25172742

Nguyen, Phi-Hung; Zhao, Bing Tian; Lee, Jeong Hyung; Kim, Young Ho; Min, Byung Sun; Woo, Mi Hee

2015-02-01

260

Alpha-lipoic Acid prevents endotoxic shock and multiple organ dysfunction syndrome induced by endotoxemia in rats.  

PubMed

Alpha-lipoic acid (ALA), a naturally occurring disulfide derivative of octanoic acid, serves as a strong antioxidant and has been reported to possess anti-inflammatory effects. The aim of the present study is to investigate the preventive and therapeutic effects of ALA on multiple organ dysfunction syndrome (MODS) caused by endotoxemia in rats. Male Wistar rats were intravenously infused with lipopolysaccharide (LPS) (10 mg/kg) to induce endotoxemia. Alpha-lipoic acid 10, 20, or 40 mg/kg was administered intravenously 60 min before (pretreatment) LPS challenge, and ALA 40 mg/kg was administered intravenously 30 min after (posttreatment) LPS challenge. Pretreatment and posttreatment with ALA significantly improved the deleterious hemodynamic changes 8 h after LPS challenge, including hypotension and bradycardia. Alpha-lipoic acid reduced the plasma levels of glutamic pyruvic transaminase, blood urea nitrogen, lactate dehydrogenase, tumor necrosis factor-?, nitric oxide metabolites, and thrombin-antithrombin complex, which increased markedly after LPS challenge. The induction of inducible nitric oxide synthase both in the liver and the lung and vascular superoxide anion production were also significantly suppressed by ALA. Moreover, ALA significantly attenuated LPS-induced caspase-3 activation in cardiomyocytes and improved survival rate. In conclusion, ALA effectively attenuated LPS-induced acute inflammatory response and improved MODS. The antioxidant and anti-inflammatory effects of ALA may contribute to these beneficial effects. Alpha-lipoic acid might be considered as a novel therapeutic strategy in the prevention of sepsis-induced MODS and inflammatory vascular diseases. PMID:25514429

Shen, Hsin-Hsueh; Lam, Kwok-Keung; Cheng, Pao-Yun; Kung, Ching-Wen; Chen, Shu-Ying; Lin, Pei-Chiang; Chung, Ming-Ting; Lee, Yen-Mei

2015-04-01

261

Synergistic Cytokine-Induced Nitric Oxide Production in Human Alveolar Epithelial Cells  

E-print Network

Synergistic Cytokine-Induced Nitric Oxide Production in Human Alveolar Epithelial Cells Soonjo Kwon IL-1 , TNF- , and IFN- in NO produc- tion from human alveolar epithelial cells (A549) are proposed rights of reproduction in any form reserved. #12;demonstrated that a human Type II alveolar epithelial

George, Steven C.

262

Mechanisms of Synergistic Cytokine-Induced Nitric Oxide Production in Human Alveolar Epithelial Cells  

E-print Network

Mechanisms of Synergistic Cytokine-Induced Nitric Oxide Production in Human Alveolar Epithelial NO production in human alveolar epithelial cells in a synergistic (nonlinear or nonadditive) mannerNOS protein, and NO production in A549 monolayers (human alveolar epithelial cell line) in response

George, Steven C.

263

Cystic Fibrosis Transmembrane Conductance Regulator-Dependent Regulation of Epithelial Inducible Nitric Oxide Synthase Expression  

Microsoft Academic Search

Recent evidence has shown that the inducible form of nitric oxide (NO) synthase (NOS2) has reduced ex- pression in airway epithelia of patients with cystic fibrosis (CF) despite the presence of chronic inflamma- tion. The goal of this paper is to determine whether NOS2 expression is regulated by the presence of func- tional CF transmembrane conductance regulator (CFTR). Using a

Wendy K. Steagall; Heather L. Elmer; Kristine G. Brady; Thomas J. Kelley

2000-01-01

264

Long-Term Treatment of Anterior Pituitary Cells with Nitric Oxide Induces Programmed Cell Death  

Microsoft Academic Search

Nitric oxide (NO) plays a complex role in modulating pro- grammed cell death. It can either protect the cell from apo- ptotic death or mediate apoptosis, depending on its concen- tration and the cell type and\\/or status. In this study, we demonstrate that long-term exposition to NO induces cell death of anterior pituitary cells from Wistar female rats. DETA NONOate

MIGUEL OMAR VELARDEZ; ARIEL HERNAN POLIANDRI; JIMENA PAULA CABILLA; CARLOS ARMANDO BODO; LETICIA INES MACHIAVELLI; BEATRIZ HAYDEEDUVILANSKI

2003-01-01

265

Endotoxin-Induced Uveitis in the Rat Is Attenuated by Inhibition of Nitric Oxide Production  

Microsoft Academic Search

Purpose. These experiments were undertaken to assess the role of increased nitric oxide pro- duction in the pathogenesis of vascular dysfunction associated with endotoxin-induced uveitis. Methods. Lipopolysaccharides (LPS) (100 ng of Salmonella Minnesota) was injected into foot- pads of Lewis rats randomly assigned to an untreated group or to a group treated with subcuta- neous injections of aminoguanidine, a selective

Ronald G. Tilton; Kathy Chang; John A. Corbett; Thomas P. Misko; Mark G. Currie; Nalini S. Bora; Henry J. Kaplan; Joseph R. Williamson

1994-01-01

266

Bronchoconstriction Induced by Citric Acid Inhalation in Guinea Pigs Role of Tachykinins, Bradykinin, and Nitric Oxide  

Microsoft Academic Search

Gastroesophageal acid reflux into the airways can trigger asthma attacks. Indeed, citric acid inhala- tion causes bronchoconstriction in guinea pigs, but the mechanism of this effect has not been fully clarified. We investigated the role of tachykinins, bradykinin, and nitric oxide (NO) on the citric acid- induced bronchoconstriction in anesthetized and artificially ventilated guinea pigs. Citric acid inhala- tion (2-20

FABIO L. M. RICCIARDOLO; VANDA RADO; LEONARDO M. FABBRI; PETER J. STERK; GIUSEPPE U. DI; PIERANGELO GEPPETTI

267

The flavonoid luteolin induces nitric oxide production and arterial relaxation  

PubMed Central

Purpose Luteolin, a flavone present in many foods and medicinal plants, may have beneficial effects on various human chronic diseases. In the present study, we investigated the hypothesis that luteolin can directly act on vascular endothelial cells (ECs), leading to nitric oxide (NO) production and subsequent vascular relaxation. Methods Rat aortic rings were mounted in organ bath. Luteolin was added cumulatively and vessel relaxation of rat aortic rings precontracted with phenylephrine (PE) or potassium was recorded. Endothelial nitric oxide synthase (eNOS) phosphorylation at Ser1177 and NO production from aortic rings and primary human aortic endothelial cells (HAECs) exposed to luteolin were measured by using Western blot and fluorometric assay, respectively. Results Luteolin dose-dependently (10-100 ?mol/L) elicited relaxation of PE- or potassium-contracted aortic rings. The vasorelaxation effect of luteolin was attenuated by the eNOS inhibitor, N-nitro-L-arginine methyl ester, suggesting that this luteolin action is at least partially mediated by activating eNOS activity. We further found that luteolin dose-dependently (10-100 ?mol/L) increased eNOS phosphorylation at Ser1177 (up to 1.9 fold) in isolated rat rings. Consistently, exposure of HAECs to luteolin also increased eNOS phosphorylation and NO production. Conclusion Luteolin may be a vascular protective agent by directly acting on vascular ECs to stimulate NO-dependent vascular dilatation. PMID:23604495

Si, Hongwei; Wyeth, Richard P.; Liu, Dongmin

2013-01-01

268

Anmindenols A and B, inducible nitric oxide synthase inhibitors from a marine-derived Streptomyces sp.  

PubMed

Anmindenols A (1) and B (2), inhibitors of inducible nitric oxide synthase (iNOS), were isolated from a marine-derived bacterium Streptomyces sp. Their chemical structures were elucidated by interpreting various spectroscopic data, including IR, MS, and NMR. Anmindenols A and B are sesquiterpenoids possessing an indene moiety with five- and six-membered rings derived from isoprenyl units. The absolute configuration of C-4 in anmindenol B was determined by electronic circular dichroism (ECD) of a dimolybdenum complex. Anmindenols A (1) and B (2) inhibited nitric oxide production in stimulated RAW 264.7 macrophage cells with IC50 values of 23 and 19 ?M, respectively. PMID:24878306

Lee, Jihye; Kim, Hiyoung; Lee, Tae Gu; Yang, Inho; Won, Dong Hwan; Choi, Hyukjae; Nam, Sang-Jip; Kang, Heonjoong

2014-06-27

269

Nitric oxide enhances MPP(+)-induced hydroxyl radical generation via depolarization activated nitric oxide synthase in rat striatum.  

PubMed

We examined the effect of N(G)-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase (NOS) inhibitor, on extracellular potassium ion concentration ([K(+)](o))-enhanced hydroxyl radical (.OH) generation due to 1-methyl-4-phenylpyridinium ion (MPP(+)) was examined in the rat striatum. Rats were anesthetized, and sodium salicylate in Ringer's solution (0.5 nmol/microl per min) was infused through a microdialysis probe to detect the generation of.OH as reflected by the non-enzymatic formation of 2,3-dihydroxybenzoic acid (DHBA) in the striatum. Induction of KCl (20, 70 and 140 mM) increased MPP(+)-induced.OH formation trapped as 2,3-dihydroxybenzoic acid (DHBA) in a concentration dependent manner. However, the application of L-NAME (5 mg/kg i.v.) abolished the [K(+)](o) depolarization-induced.OH formation with MPP(+). Dopamine (DA; 10 microM) also increased the levels of DHBA due to MPP(+). However, the effect of DA after application of L-NAME did not change the levels of DHBA. On the other hand, the application of allopurinol (20 mg/kg i.v., 30 min prior to study), a xanthine oxidase (XO) inhibitor was abolished the both [K(+)](o)- and DA-induced.OH generation. Moreover, when iron(II) was administered to MPP(+) then [K(+)](o) (70 mM)-pretreated animals, a marked increase in the level of DHBA. However, when corresponding experiments were performed with L-NAME-pretreated animals, the same results were obtained. Therefore, NOS activation may be no relation to Fenton-type reaction via [K(+)](o) depolarization-induced.OH generation. The present results suggest that [K(+)](o)-induced depolarization augmented MPP(+)-induced.OH formation by enhancing NO synthesis. PMID:11384616

Obata, T; Yamanaka, Y

2001-06-01

270

Inducible nitric oxide synthase expression is reduced in cystic fibrosis murine and human airway epithelial cells.  

PubMed Central

It has been reported that exhaled nitric oxide levels are reduced in cystic fibrosis (CF) patients. We have examined the inducible isoform of nitric oxide synthase (iNOS) in the airways by immunostaining and found that iNOS is constitutively expressed in the airway epithelia of non-CF mouse and human tissues but essentially absent in the epithelium of CF airways. We explored potential consequences of lost iNOS expression and found that iNOS inhibition significantly increases mouse nasal trans-epithelial potential difference, and hindered the ability of excised mouse lungs to prevent growth of Pseudomonas aeruginosa. The absence of continuous nitric oxide production in epithelial cells of CF airways may play a role in two CF-associated characteristics: hyperabsorption of sodium and susceptibility to bacterial infections. PMID:9739054

Kelley, T J; Drumm, M L

1998-01-01

271

Nitric Oxide Donor SNAP Induces Apoptosis in Smooth Muscle Cells through cGMP-Independent Mechanism  

Microsoft Academic Search

Recent evidence suggests that nitric oxide (NO) may function as a second messenger in the intracellular signal transduction pathways. We explored the possibility that NO was involved in the signal for triggering apoptosis in smooth muscle cells (SMCs). Chemical NO donors induced SMCs apoptosis in a concentration- and time-dependent manner. The membrane-permeable cGMP analogue, dibutyryl-cGMP, did not induce SMCs apoptosis,

E. Nishio; K. Fukushima; M. Shiozaki; Y. Watanabe

1996-01-01

272

Cytokine and Nitric Oxide Production in the Acute Phase of Bacterial Cell Wall-Induced Arthritis  

Microsoft Academic Search

We have investigated the temporal relationship among proinflammatory cytokine expression, nitric oxide (NO) production and joint inflammation in the acute phase of bacterial cell wall-derived peptidoglycan polysaccharide(PG\\/PS)-induced arthritis. Acute joint inflammation was induced in female LEW\\/N rats by a single intraperitoneal injection of PG\\/PS. Arthritis index and paw volume were quantified and joint histopathology was evaluated during acute joint inflammation

John W. Fuseler; Elaine M. Conner; Jonathan M. Davis; Robert E. Wolf; Matthew B. Grisham

1997-01-01

273

Expression of inducible nitric oxide synthase and nitrotyrosine in colonic epithelium in inflammatory bowel disease  

Microsoft Academic Search

BACKGROUND & AIMS: Inducible nitric oxide synthase (iNOS) is generated in several cell types by treatment with lipopolysaccharides or cytokines. Earlier studies suggested that ulcerative colitis is associated with increased NO produced by iNOS; however, the cellular source of the NO synthesis was not identified. A possible mechanism of NO-induced cellular damage is through its interaction with superoxide to produce

II Singer; DW Kawka; S Scott; JR Weidner; RA Mumford; TE Riehl; WF Stenson

1996-01-01

274

Regulatory Effect of 25-hydroxyvitamin D3 on Nitric Oxide Production in Activated Microglia  

PubMed Central

Microglia are activated by inflammatory and pathophysiological stimuli in neurodegenerative diseases, and activated microglia induce neuronal damage by releasing cytotoxic factors like nitric oxide (NO). Activated microglia synthesize a significant amount of vitamin D3 in the rat brain, and vitamin D3 has an inhibitory effect on activated microglia. To investigate the possible role of vitamin D3 as a negative regulator of activated microglia, we examined the effect of 25-hydroxyvitamin D3 on NO production of lipopolysaccharide (LPS)-stimulated microglia. Treatment with LPS increased the production of NO in primary cultured and BV2 microglial cells. Treatment with 25-hydroxyvitamin D3 inhibited the generation of NO in LPS-activated primary microglia and BV2 cells. In addition to NO production, expression of 1-?-hydroxylase and the vitamin D receptor (VDR) was also upregulated in LPS-stimulated primary and BV2 microglia. When BV2 cells were transfected with 1-?-hydroxylase siRNA or VDR siRNA, the inhibitory effect of 25-hydroxyvitamin D3 on activated BV2 cells was suppressed. 25-Hydroxyvitamin D3 also inhibited the increased phosphorylation of p38 seen in LPS-activated BV2 cells, and this inhibition was blocked by VDR siRNA. The present study shows that 25-hydroxyvitamin D3 inhibits NO production in LPS-activated microglia through the mediation of LPS-induced 1-?-hydroxylase. This study also shows that the inhibitory effect of 25-hydroxyvitamin D3 on NO production might be exerted by inhibiting LPS-induced phosphorylation of p38 through the mediation of VDR signaling. These results suggest that vitamin D3 might have an important role in the negative regulation of microglial activation. PMID:25352759

Hur, Jinyoung; Lee, Pyeongjae; Kim, Mi Jung

2014-01-01

275

Effect of quercetin on the production of nitric oxide in murine macrophages stimulated with lipopolysaccharide from Prevotella intermedia  

PubMed Central

Purpose Nitric oxide (NO) is a short-lived bioactive molecule that is known to play an important role in the pathogenesis of periodontal disease. In the current study, we investigated the effect of the flavonoid quercetin on the production of NO in murine macrophages activated with lipopolysaccharide (LPS) from Prevotella intermedia, a pathogen related to inflammatory periodontal disease, and tried to elucidate the underlying mechanisms of action. Methods LPS was isolated from P. intermedia ATCC 25611 cells by the standard hot phenol-water method. The concentration of NO in cell culture supernatants was determined by measuring the accumulation of nitrite. Inducible NO synthase (iNOS) and heme oxygenase-1 (HO-1) protein expression, phosphorylation of c-Jun N-terminal kinase (JNK) and p38, inhibitory ?B (I?B)-? degradation, and signal transducer and activator of transcription 1 (STAT1) phosphorylation were analyzed via immunoblotting. Results Quercetin significantly attenuated iNOS-derived NO production in RAW246.7 cells activated by P. intermedia LPS. In addition, quercetin induced HO-1 protein expression in cells activated with P. intermedia LPS. Tin protoporphyrin IX (SnPP), a competitive inhibitor of HO-1, abolished the inhibitory effect of quercetin on LPS-induced NO production. Quercetin did not affect the phosphorylation of JNK and p38 induced by P. intermedia LPS. The degradation of I?B-? induced by P. intermedia LPS was inhibited when the cells were treated with quercetin. Quercetin also inhibited LPS-induced STAT1 signaling. Conclusions Quercetin significantly inhibits iNOS-derived NO production in murine macrophages activated by P. intermedia LPS via anti-inflammatory HO-1 induction and inhibition of the nuclear factor-?B and STAT1 signaling pathways. Our study suggests that quercetin may contribute to the modulation of host-destructive responses mediated by NO and appears to have potential as a novel therapeutic agent for treating inflammatory periodontal disease. PMID:24040572

Cho, Yun-Jung

2013-01-01

276

Traumatic Brain Injury Disrupts Cerebrovascular Tone Through Endothelial Inducible Nitric Oxide Synthase Expression and Nitric Oxide Gain of Function  

PubMed Central

Background Traumatic brain injury (TBI) has been reported to increase the concentration of nitric oxide (NO) in the brain and can lead to loss of cerebrovascular tone; however, the sources, amounts, and consequences of excess NO on the cerebral vasculature are unknown. Our objective was to elucidate the mechanism of decreased cerebral artery tone after TBI. Methods and Results Cerebral arteries were isolated from rats 24 hours after moderate fluid?percussion TBI. Pressure?induced increases in vasoconstriction (myogenic tone) and smooth muscle Ca2+ were severely blunted in cerebral arteries after TBI. However, myogenic tone and smooth muscle Ca2+ were restored by inhibition of NO synthesis or endothelium removal, suggesting that TBI increased endothelial NO levels. Live native cell NO, indexed by 4,5?diaminofluorescein (DAF?2 DA) fluorescence, was increased in endothelium and smooth muscle of cerebral arteries after TBI. Clamped concentrations of 20 to 30 nmol/L NO were required to simulate the loss of myogenic tone and increased (DAF?2T) fluorescence observed following TBI. In comparison, basal NO in control arteries was estimated as 0.4 nmol/L. Consistent with TBI causing enhanced NO?mediated vasodilation, inhibitors of guanylyl cyclase, protein kinase G, and large?conductance Ca2+?activated potassium (BK) channel restored function of arteries from animals with TBI. Expression of the inducible isoform of NO synthase was upregulated in cerebral arteries isolated from animals with TBI, and the inducible isoform of NO synthase inhibitor 1400W restored myogenic responses following TBI. Conclusions The mechanism of profound cerebral artery vasodilation after TBI is a gain of function in vascular NO production by 60?fold over controls, resulting from upregulation of the inducible isoform of NO synthase in the endothelium. PMID:25527626

Villalba, Nuria; Sonkusare, Swapnil K.; Longden, Thomas A.; Tran, Tram L.; Sackheim, Adrian M.; Nelson, Mark T.; Wellman, George C.; Freeman, Kalev

2014-01-01

277

Effects of nitric oxide on cholesterol metabolism in macrophages.  

PubMed

Nitric oxide (NO) is associated with atherogenic process by inhibiting the proliferation of vascular smooth muscle cells, adhesion of monocyte/macrophages, aggregation and adhesion of platelets and oxidation of LDL, but it is not clear whether NO affects cellular cholesterol metabolism or not. We investigated cholesterol metabolism in murine macrophages (J774A.1) by regulating NO production. Incubation with S-nitroso-N-acetylpenicillamine (SNAP), an NO donor, had no influence on cellular cholesterol accumulation induced by LDL or acetylated LDL (acetyl-LDL). Lipopolysaccharide (LPS) stimulated NO production in a dose-dependent manner in the presence of LDL or acetyl-LDL but did not change LDL-induced cellular cholesterol accumulation. In the presence of acetyl-LDL, LPS stimulated NO production but significantly inhibited cholesteryl ester accumulation in a dose-dependent manner (37.7% decrease by 100 micrograms/ml of LPS), but LPS simulation did not change free cholesterol content. NG-monomethyl-L-arginine (L-NMMA), inhibitor of NO synthase, suppressed NO production and addition of L-arginine restored NO production, but these regulations did not alter LPS-induced esterified cholesterol reduction. These results suggest that NO generation in atherosclerotic lesions does not influence cholesterol metabolism in macrophages. PMID:9105561

Shimizu, H; Taniguchi, T; Ishikawa, Y; Yokoyama, M

1997-03-21

278

Exercise training reverses age-induced inducible nitric oxide synthase upregulation  

E-print Network

homeostasis and prevent free radical-related damage. However, at high levels, chronic production of ?NO represents a formidable vehicle for pathology. Reid (135) suggested a biphasic response for nitric oxide on Ca2+ release dependent on nitric oxide... concentration and time of exposure. Recent studies indicated that overproduction of nitric oxide may be involved in muscle wasting, or cachexia, consistently observed with chronic heart failure, cancer, AIDS, and sepsis (2, 3, 29, 65). Elevated iNOS levels...

Song, Wook

2005-02-17

279

The effects of oxidized low density lipoproteins on inducible mouse macrophage gene expression are gene and stimulus dependent.  

PubMed Central

Oxidized LDL has been previously reported to suppress the expression of genes induced in mononuclear phagocytes by inflammatory stimuli. In this study we extend these findings to demonstrate that the suppressive effects of oxidized LDL vary depending upon the gene being monitored and the stimulus being used to induce or enhance its expression. The expression of a selection of LPS-inducible genes exhibited differential sensitivity to pretreatment with oxidized LDL. Furthermore, the ability of oxidized LDL to suppress gene expression varied markedly with the inducing stimulus used. TNF alpha and IP-10 mRNA expression induced by IFN gamma and IL-2 was markedly more sensitive to suppression by oxidized LDL than that induced by LPS. The cooperative effects of IFN gamma and LPS on the expression of the inducible nitric oxide synthase gene were suppressed by oxidized LDL while the antagonistic effect of IFN gamma on LPS-induced expression of the TNF receptor type II mRNA was not altered. The suppressive activity of LDL was acquired only after extensive oxidation and was localized in the extractable lipid component. These results suggest a potent and direct connection between the oxidative modification of LDL and the chronic inflammation seen in atherogenic lesions. Furthermore, the appreciable selectivity of oxidized LDL in mediating secondary control of cytokine gene expression demonstrates that the active material(s) is targeted to disrupt specific intracellular signaling pathways. Images PMID:7537753

Hamilton, T A; Major, J A; Chisolm, G M

1995-01-01

280

Testosterone production in mice lacking inducible nitric oxide synthase expression is sensitive to restraint stress.  

PubMed

Immobilization stress (IMO) induces a rapid increase in glucocorticoid secretion [in rodents, corticosterone CORT)] and this is associated with decreased circulating testosterone (T) levels. Nitric oxide (NO), a reactive free radical and neurotransmitter, has been reported to be produced at higher rates in tissues such as brain during stress. The biosynthesis of T is also known to be dramatically suppressed by NO. Specifically, the inducible isoform of nitric oxide synthase (iNOS) was directly implicated in this suppression. To assess the respective roles of CORT and NO in stress-mediated inhibition of T production, adult wild-type (WT) and inducible nitric oxide synthase knockout (iNOS(-/-)) male mice were evaluated. Animals of each genotype were assigned to either basal control or 3-h IMO groups. Basal plasma and testicular T levels were equivalent in both genotypes, whereas testicular weights of mutant mice were significantly higher compared with WT animals. Exposure to 3-h IMO increased plasma CORT and decreased T concentrations in mice of both genotypes. Testicular T levels were also affected by stress in WT and mutant males, being sharply reduced in both genotypes. However, the concentrations of nitrite and nitrate, the stable metabolites of NO measured in testicular extracts, did not differ between control and stressed WT and iNOS(-/-) mice. These results support the hypothesis that CORT, but not NO, is a plausible candidate to mediate rapid stress-induced suppression of Leydig cell steroidogenesis. PMID:17032928

Weissman, Ben A; Sottas, Chantal M; Zhou, Ping; Iadecola, Costantino; Hardy, Matthew P

2007-02-01

281

Clozapine protects dopaminergic neurons from inflammation-induced damage by inhibiting microglial overactivation  

PubMed Central

Increasing evidence suggests a possible involvement of neuroinflammation in some psychiatric disorders, and also pharmacological reports indicate that anti-inflammatory effects are associated with therapeutic actions of psychoactive drugs, such as anti-depressants and antipsychotics. The purpose of this study was to explore whether clozapine, a widely used antipsychotic drugs, displays anti-inflammatory and neuroprotective effects. Using primary cortical and mesencephalic neuron-glia cultures, we found that clozapine was protective against inflammation-related neurodegeneration induced by lipopolysaccharide (LPS). Pretreatment of cortical or mesencephalic neuron–glia cultures with clozapine (0.1 or 1µM) for 24 hrs attenuated LPS-induced neurotoxicity. Clozapine also protected neurons against 1-methyl-4-phenylpyridinium+ (MPP+)-induced neurotoxicity, but only in cultures containing microglia, indicating an indispensable role of microglia in clozapine-afforded neuroprotection. Further observation revealed attenuated LPS-induced microglial activation in primary neuron-glia cultures and in HAPI microglial cell line with clozapine pretreatment. Clozapine ameliorated the production of microglia-derived superoxide and intracellular reactive oxygen species (ROS), as well as the production of nitric oxide and TNF-? following LPS. In addition, the protective effect of clozapine was not observed in neuron-glia cultures from mice lacking functional NADPH oxidase (PHOX), a key enzyme for superoxide production in immune cells. Further mechanistic studies demonstrated that clozapine pretreatment inhibited LPS-induced translocation of cytosolic subunit p47phox to the membrane in microglia, which was most likely though inhibiting the phosphoinositide 3-kinase (PI3K) pathway. Taken together, this study demonstrates that clozapine exerts neuroprotective effect via the attenuation of microglia activation through inhibition of PHOX-generated ROS production and suggests potential use of antipsychotic drugs for neuroprotection. PMID:21870076

Hu, Xiaoming; Zhou, Hui; Zhang, Dan; Yang, Sufen; Qian, Li; Wu, Hung-Ming; Chen, Po-See; Wilson, Belinda; Gao, Hui-Ming; Lu, Ru-band; Hong, Jau-Shyong

2013-01-01

282

The role of nitric oxide in testosterone-induced vasodilation in pig coronary arteries and rat thoracic aorta  

E-print Network

Several studies have provided evidence that the administration of testosterone to vascular tissue causes vasodilation (Costarella, Yue). This study examines the role of nitric oxide (NO) as a potential mechanism of testosterone-induced vasodilation...

Piefer, Jason William

2013-02-22

283

Immunomodulatory Effect of Chinese Herbal Medicine Formula Sheng-Fei-Yu-Chuan-Tang in Lipopolysaccharide-Induced Acute Lung Injury Mice  

PubMed Central

Traditional Chinese medicine formula Sheng-Fei-Yu-Chuan-Tang (SFYCT), consisting of 13 medicinal plants, was used to treat patients with lung diseases. This study investigated the immunoregulatory effect of SFYCT on intratracheal lipopolysaccharides- (LPS-) challenged acute lung injury (ALI) mice. SFYCT attenuated pulmonary edema, macrophages, and neutrophils infiltration in the airways. SFYCT decreased inflammatory cytokines, including tumor necrosis factor-? (TNF?), interleukin-1?, and interleukin-6 and inhibited nitric oxide (NO) production but increased anti-inflammatory cytokines, interleukin-4, and interleukin-10, in the bronchoalveolar lavage fluid of LPS-challenged mice. TNF? and monocyte chemotactic protein-1 mRNA expression in the lung of LPS-challenged mice as well as LPS-stimulated lung epithelial cell and macrophage were decreased by SFYCT treatment. SFYCT treatment also decreased the inducible nitric oxide synthase expression and phosphorylation of nuclear factor-?B (NF-?B) in the lung of mice and macrophage with LPS stimulation. SFYCT treatment dose dependently decreased the LPS-induced NO and reactive oxygen species generation in LPS-stimulated macrophage. In conclusion, SFYCT attenuated lung inflammation during LPS-induced ALI through decreasing inflammatory cytokines production while increasing anti-inflammatory cytokines production. The immunoregulatory effect of SFYCT is related to inhibiting NF-?B phosphorylation. PMID:23997804

Lin, Chia-Hung; Yeh, Ching-Hua; Wang, Shulhn-Der; Wang, Jen-Shu; Kao, Shung-Te

2013-01-01

284

Extract of Meretrix meretrix Linnaeus induces angiogenesis in vitro and activates endothelial nitric oxide synthase  

NASA Astrophysics Data System (ADS)

Meretrix meretrix Linnaeus has long been used as traditional Chinese medicine in oriental medicine. The angiogentic activity of the extract of M. meretrix was investigated in this study, using human umbilical vein endothelial cells (HUVECs). Extract of M. meretrix Linnaeus (AFG-25) was prepared with acetone and ethanol precipitation, and further separated by Sephadex G-25 column. The results show that AFG-25 promoted proliferation, migration, and capillary-like tube formation in HUVECs, and in the presence of eNOS inhibitor NMA, the tube formation induced by AFG-25 is inhibited significantly. Moreover, AFG-25 could also promote the activation of endothelial nitric oxide synthase (eNOS) and the resultant elevation of nitric oxide (NO) production. The results suggested that M. meretrix contains active ingredients with angiogentic activity and eNOS/NO signal pathway is in part involved in the proangiogenesis effect induced by AFG-25.

Liu, Ming; Wei, Jianteng; Wang, Hui; Ding, Lili; Zhang, Yuyan; Lin, Xiukun

2012-09-01

285

INTERLEUKIN1?-INDUCED NITRIC OXIDE PRODUCTION AND INHIBITION OF INSULIN SECRETION IN RAT ISLETS OF LANGERHANS IS DEPENDENT UPON THE NITRIC OXIDE SYNTHASE COFACTOR TETRAHYDROBIOPTERIN  

Microsoft Academic Search

Interleukin (IL)-1?-induced inhibition of glucose-stimulated insulin secretion in rat islets of Langerhans is mediated in part by nitric oxide (NO). The NO synthase cofactor 5,6,7,8-tetrahydrobiopterin (BH4) supports NO synthesis in many cell types and IL-1?-induced NO generation and inhibition of insulin secretion have been previously correlated with intracellular BH4 levels in rat insulinoma cells. Using rat islets and the beta

P. Stickings; J. M. Cunningham

2002-01-01

286

Differential expression and induction of mRNAs encoding two inducible nitric oxide synthases in rat kidney  

Microsoft Academic Search

Differential expression and induction of mRNAs encoding two inducible nitric oxide synthases in rat kidney. We used quantitative PCR methods and renal microdissection to characterize the expression of inducible nitric oxide synthase (iNOS) mRNAs in rat kidney and cultured glomerular mesangial cells. A partial cDNA homologous to murine macrophage iNOS (macNOS), but distinct from rat vascular smooth muscle iNOS (vsmNOS),

Markus G Mohaupt; John L Elzie; Kyu Y Ahn; William L Clapp; Christopher S Wilcox; Bruce C Kone

1994-01-01

287

Nitric oxide protects anterior pituitary cells from cadmium-induced apoptosis  

Microsoft Academic Search

Cadmium (Cd2+) is a potent toxic metal for both plants and animals. Chronic exposure to low doses of Cd2+ results in damage to several organs. We have previously reported that Cd2+ induces apoptosis in anterior pituitary cells by a caspase- and oxidative stress-dependent mechanism. Nitric oxide (NO) synthesis is affected by Cd2+ in several systems. NO has been shown to

Ariel H. B. Poliandri; Miguel O. Velardez; Jimena P. Cabilla; Cristian C. A. Bodo; Leticia I. Machiavelli; Alnilan F. Quinteros; Beatriz H. Duvilanski

2004-01-01

288

Early effects of diabetes on inducible nitric oxide synthase in the kidney  

Microsoft Academic Search

NO may be responsible for the glomerular hyperfiltration observed in diabetic kidney by inducing vasodilation of the afferent\\u000a arteriole. The aim of this study was to evaluate which isoform of nitric oxide synthase (NOS) is responsible for increased\\u000a renal production of NO in diabetic kidney. Thirty male WKY rats were divided into 6 groups. Five rats were sacrificed immediately,\\u000a five

A. Cosenzi; E. Bernobich; M. Bonavita; R. Trevisan; G. Bellini; L. Campanacci

2002-01-01

289

Bifunctional effects of fucoidan on the expression of inducible nitric oxide synthase  

Microsoft Academic Search

Algal fucoidan is a marine sulfated polysaccharide with a wide variety of biological activities including anti-thrombotic and anti-inflammatory effects. This study evaluated the effect of fucoidan on the expression of inducible nitric oxide synthase (iNOS) in a macrophage cell line, RAW264.7. Low concentration range of fucoidan (10?g\\/ml) increased the basal expression level of iNOS in quiescent macrophages. However, we found

Jin Won Yang; Se Young Yoon; Soo Jin Oh; Sang Kyum Kim; Keon Wook. Kang

2006-01-01

290

Pulmonary Nitric Oxide Metabolism Following Infrarenal AorticCross-clamp-induced Ischaemia–Reperfusion Injury  

Microsoft Academic Search

Objectives to investigate endogenous pulmonary nitric oxide metabolism following infrarenal aortic cross-clamp-induced ischaemia–reperfusion injury. Methods groups of male Wistar rats (n=6) were subjected to 60 minutes of infrarenal aortic cross-clamping under general anaesthesia. Rats were culled after 0, 60 and 120 minutes» reperfusion, following release of the aortic clamp. A sham-operated control group was also studied. Acute lung injury (ALI)

R. Pararajasingam; S. C. Weight; P. R. F. Bell; M. L. Nicholson; R. D. Sayers

2000-01-01

291

Impairment of nitric oxide-mediated relaxations in anaesthetized autoperfused streptozotocin-induced diabetic rats  

Microsoft Academic Search

This work was designed to determine in vivo the influence of the metabolic control of streptozotocin-induced diabetic rats,\\u000a measured by the levels of haemoglobin glycosylation in blood (HbA1c), on developing vascular endothelial dysfunction. For this, the vasoactive responses to basal and stimulated endothelial\\u000a nitric oxide (NO) were studied using the technique of the anaesthetized autoperfused rat, analyzing the responses to

Javier Angulo; Leocadio Rodríguez-Mañas; Concepción Peiró; Marta Neira; Jesús Marín; Carlos F. Sánchez-Ferrer

1998-01-01

292

Acute pancreatitis, expression of inducible nitric oxide synthase and defective insulin secretion  

Microsoft Academic Search

A high level of nitric oxide (NO) produced by inducible NO synthase (iNOS) is involved in pancreatic beta-cell dysfunction and apoptosis. In the present study, we examined whether iNOS is also expressed in beta cells after induction of acute pancreatitis (AP) in the rat. Pancreatic islets taken from AP animals and incubated for 60 min in the presence of 20.0 mmol\\/l glucose

Saleem S. Qader; Mats Ekelund; Roland Andersson; Stefanie Obermuller; Albert Salehi

2003-01-01

293

Methamphetamine-induced nitric oxide promotes vesicular transport in blood-brain barrier endothelial cells.  

PubMed

Methamphetamine's (METH) neurotoxicity is thought to be in part due to its ability to induce blood-brain barrier (BBB) dysfunction. Here, we investigated the effect of METH on barrier properties of cultured rat primary brain microvascular endothelial cells (BMVECs). Transendothelial flux doubled in response to METH, irrespective of the size of tracer used. At the same time, transendothelial electrical resistance was unchanged as was the ultrastructural appearance of inter-endothelial junctions and the distribution of key junction proteins, suggesting that METH promoted vesicular but not junctional transport. Indeed, METH significantly increased uptake of horseradish peroxidase into vesicular structures. METH also enhanced transendothelial migration of lymphocytes indicating that the endothelial barrier against both molecules and cells was compromised. Barrier breakdown was only observed in response to METH at low micromolar concentrations, with enhanced vesicular uptake peaking at 1 ?M METH. The BMVEC response to METH also involved rapid activation of endothelial nitric oxide synthase and its inhibition abrogated METH-induced permeability and lymphocyte migration, indicating that nitric oxide was a key mediator of BBB disruption in response to METH. This study underlines the key role of nitric oxide in BBB function and describes a novel mechanism of drug-induced fluid-phase transcytosis at the BBB. PMID:22960442

Martins, Tânia; Burgoyne, Thomas; Kenny, Bridget-Ann; Hudson, Natalie; Futter, Clare E; Ambrósio, António F; Silva, Ana P; Greenwood, John; Turowski, Patric

2013-02-01

294

Auxin-induced nitric oxide, cGMP and gibberellins were involved in the gravitropism  

NASA Astrophysics Data System (ADS)

Gravitropism is the asymmetric growth or curvature of plant organs in response to gravistimulation. There is a complex signal transduction cascade which involved in the differential growth of plants in response to changes in the gravity vector. The role of auxin in gravitropism has been demonstrated by many experiments, but little is known regarding the molecular details of such effects. In our studies before, mediation of the gravitropic bending of soybean roots and rice leaf sheath bases by nitric oxide, cGMP and gibberellins, are induced by auxin. The asymmetrical distribution of nitric oxide, cGMP and gibberellins resulted from the asymmetrical synthesis of them in bending sites. In soybean roots, inhibitions of NO and cGMP synthesis reduced differential NO and cGMP accumulation respectively, which both of these effects can lead to the reduction of gravitropic bending. Gibberellin-induced OsXET, OsEXPA4 and OsRWC3 were also found involved in the gravitropic bending. These data indicated that auxin-induced nitric oxide, cGMP and gibberellins were involved in the gravitropism. More experiments need to prove the more detailed mechanism of them.

Cai, Weiming; Hu, Liwei; Hu, Xiangyang; Cui, Dayong; Cai, Weiming

295

Methamphetamine-induced nitric oxide promotes vesicular transport in blood–brain barrier endothelial cells  

PubMed Central

Methamphetamine's (METH) neurotoxicity is thought to be in part due to its ability to induce blood–brain barrier (BBB) dysfunction. Here, we investigated the effect of METH on barrier properties of cultured rat primary brain microvascular endothelial cells (BMVECs). Transendothelial flux doubled in response to METH, irrespective of the size of tracer used. At the same time, transendothelial electrical resistance was unchanged as was the ultrastructural appearance of inter-endothelial junctions and the distribution of key junction proteins, suggesting that METH promoted vesicular but not junctional transport. Indeed, METH significantly increased uptake of horseradish peroxidase into vesicular structures. METH also enhanced transendothelial migration of lymphocytes indicating that the endothelial barrier against both molecules and cells was compromised. Barrier breakdown was only observed in response to METH at low micromolar concentrations, with enhanced vesicular uptake peaking at 1 ?M METH. The BMVEC response to METH also involved rapid activation of endothelial nitric oxide synthase and its inhibition abrogated METH-induced permeability and lymphocyte migration, indicating that nitric oxide was a key mediator of BBB disruption in response to METH. This study underlines the key role of nitric oxide in BBB function and describes a novel mechanism of drug-induced fluid-phase transcytosis at the BBB. PMID:22960442

Martins, Tânia; Burgoyne, Thomas; Kenny, Bridget-Ann; Hudson, Natalie; Futter, Clare E.; Ambrósio, António F.; Silva, Ana P.; Greenwood, John; Turowski, Patric

2013-01-01

296

Lipopolysaccharide induces multinuclear cell from RAW264.7 line with increased phagocytosis activity  

SciTech Connect

Highlights: Black-Right-Pointing-Pointer LPS induces multinuclear cells from murine macrophage-derived RAW264.7 cells. Black-Right-Pointing-Pointer The multinuclear cells are formed through cell-cell fusion in the presence of Ca{sup 2+}. Black-Right-Pointing-Pointer The multinuclear cells do not express osteoclast-specific enzymes. Black-Right-Pointing-Pointer They internalized more and larger beads than mononuclear cells and osteoclasts. -- Abstract: Lipopolysaccharide (LPS), an outer membrane component of Gram-negative bacteria, induces strong proinflammatory responses, including the release of cytokines and nitric oxide from macrophage. In this study, we found that a murine macrophage-derived line, RAW264.7, became multinuclear through cell-cell fusion after incubation with highly purified LPS or synthetic lipid A in the presence of Ca{sup 2+}. The same cell line is known to differentiate into multinuclear osteoclast, which expresses a specific proton pumping ATPase together with osteoclast markers on stimulation by the extracellular domain of receptor activator of nuclear factor {kappa}B ligand (Toyomura, T., Murata, Y., Yamamoto, A., Oka, T., Sun-Wada, G.-H., Wada, Y. and Futai, M., 2003). The LPS-induced multinuclear cells did not express osteoclast-specific enzymes including tartrate-resistant acid phosphatase and cathepsin K. During multinuclear cell formation, the cells internalized more and larger polystyrene beads (diameter 6-15 {mu}m) than mononuclear cells and osteoclasts. The internalized beads were located in lysosome-marker positive organelles, which were probably phagolysosomes. The LPS-induced multinuclear cell could be a good model system to study phagocytosis of large foreign bodies.

Nakanishi-Matsui, Mayumi, E-mail: nakanim@iwate-med.ac.jp [Department of Biochemistry, Faculty of Pharmaceutical Sciences, Iwate Medical University, Futai Special Laboratory, Yahaba, Iwate 028-3694 (Japan)] [Department of Biochemistry, Faculty of Pharmaceutical Sciences, Iwate Medical University, Futai Special Laboratory, Yahaba, Iwate 028-3694 (Japan); Yano, Shio; Matsumoto, Naomi; Futai, Masamitsu [Department of Biochemistry, Faculty of Pharmaceutical Sciences, Iwate Medical University, Futai Special Laboratory, Yahaba, Iwate 028-3694 (Japan)] [Department of Biochemistry, Faculty of Pharmaceutical Sciences, Iwate Medical University, Futai Special Laboratory, Yahaba, Iwate 028-3694 (Japan)

2012-08-24

297

Inhibition of lipopolysaccharide-induced inducible nitric oxide synthase expression by a novel compound, mercaptopyrazine, through suppression of nuclear factor-kappaB binding to DNA  

Microsoft Academic Search

Macrophage cells in response to cytokines and endotoxins produced a large amount of nitric oxide (NO) by expression of inducible nitric oxide synthase (iNOS), resulting in acute or chronic inflammatory disorders including septic hypotension and atherosclerosis. In the present study, we investigated the effect and the mechanism of mercaptopyrazine (MP) in the induction of iNOS and NO production as a

Sunny Lim; Keon Wook Kang; Soo-Young Park; Seok-In Kim; Yon Sik Choi; Nak-Doo Kim; Ki-Up Lee; Hong-Kyu Lee; Youngmi Kim Pak

2004-01-01

298

Neostigmine-induced contraction and nitric oxide-induced relaxation of isolated ileum from STZ diabetic guinea pigs  

PubMed Central

Both delayed gastrointestinal transit and autonomic neuropathy have been documented in patients with diabetes mellitus. The mechanism of neostigmine, an agent that mimics release of acetylcholine from autonomic neurons by prokinetic agents, to contract smooth muscle, despite dysfunctional enteric neural pathways, was determined using isolated ilea from STZ-treated and control guinea pigs. Both bethanechol- and neostigmine-induced contractions were stronger in diabetic ileum. Bethanechol-induced contractions of control but not diabetic ileum were increased by low dose scopolamine suggesting reduced activation of presynaptic muscarinic autoreceptors in diabetic ileum. The muscarinic receptor antagonist 4-DAMP strongly, but the nicotinic receptor antagonist hexamethonium only weakly, reduced neostigmine-induced contractions of control and diabetic ilea. The amount of acetylcholine, inferred from tissue choline content, was increased in diabetic ileum. Nicotinic neural and noncholinergic postjunctional smooth muscle receptors contributed more strongly to neostigmine-induced contractions in diabetic than control ileum. Relaxation of diabetic ileum by exogenous nitric oxide generated from sodium nitroprusside was comparable to control ileum, but smooth muscle relaxation by L-arginine using neuronal nitric oxide synthase to generate nitric oxide was weaker in diabetic ileum with evidence for a role for inducible nitric oxide synthase. Despite autonomic neuropathy, neostigmine strongly contracted ileum from diabetic animals but by a different mechanism including stronger activation of postjunctional muscarinic receptors, greater synaptic acetylcholine, stronger activation of noncholinergic excitatory pathways, and weaker activation of inhibitory pathways. A selective medication targeting a specific neural pathway may more effectively treat disordered gastrointestinal transit in patients with diabetes mellitus. PMID:21880552

Cellini, Joseph; Jukic, Anne Marie Zaura; LePard, Kathy J.

2011-01-01

299

A Neurovascular Transmission Model for Acupuncture-induced Nitric Oxide  

Microsoft Academic Search

Acupuncture is the practice of inserting needles into the body to reduce pain or induce anesthesia. More broadly, acupuncture is a family of procedures involving the stimulation of anatomical locations on or in the skin by a variety of techniques. Employing acupuncture to treat human disease or maintain bodily condition has been practiced for thousands of years. However, the mechanism(s)

Sheng-Hsiung Hsiao; Li-Jen Tsai

2008-01-01

300

Ghrelin-induced growth hormone release from goldfish pituitary cells is nitric oxide dependent.  

PubMed

Ghrelin (GRLN) is an important neuroendocrine regulator of growth hormone (GH) release in vertebrates. Previous studies show goldfish (g)GRLN(19)-induced GH from the goldfish pituitary involves voltage sensitive Ca(2+) channels, increases in intracellular Ca(2+) and the PKC signalling pathway. We set out to examine the role of the nitric oxide (NO) pathway in gGLRN(19)-induced GH release from primary cultures of goldfish pituitary cells using pharmacological regulators in cell column perifusion systems. The NO scavenger PTIO abolished gGRLN(19)-induced GH release and co-treatment with the NO donor SNP and GRLN did not produce additive GH release responses. Nitric oxide synthase (NOS) inhibitors 1400 W and 7-Ni abolished GRLN-induced GH release while treatment with another NOS inhibitor, AGH, had no significant effect. Taken together, these results demonstrate that the NOS/NO is an integral component of gGRLN(19)-induced signalling within the goldfish pituitary cells, and given the relative specificity of AGH for inducible NOS and endothelial NOS isoforms, suggests that neuronal NOS is the likely NOS isoform utilized in goldfish somatotropes by this physiological regulator. PMID:22935824

Grey, Caleb L; Chang, John P

2012-11-01

301

The endogenous nitric oxide mediates selenium-induced phytotoxicity by promoting ROS generation in Brassica rapa.  

PubMed

Selenium (Se) is suggested as an emerging pollutant in agricultural environment because of the increasing anthropogenic release of Se, which in turn results in phytotoxicity. The most common consequence of Se-induced toxicity in plants is oxidative injury, but how Se induces reactive oxygen species (ROS) burst remains unclear. In this work, histofluorescent staining was applied to monitor the dynamics of ROS and nitric oxide (NO) in the root of Brassica rapa under Se(IV) stress. Se(IV)-induced faster accumulation of NO than ROS. Both NO and ROS accumulation were positively correlated with Se(IV)-induced inhibition of root growth. The NO accumulation was nitrate reductase (NR)- and nitric oxide synthase (NOS)-dependent while ROS accumulation was NADPH oxidase-dependent. The removal of NO by NR inhibitor, NOS inhibitor, and NO scavenger could alleviate Se(IV)-induced expression of Br_Rbohs coding for NADPH oxidase and the following ROS accumulation in roots, which further resulted in the amelioration of Se(IV)-induced oxidative injury and growth inhibition. Thus, we proposed that the endogenous NO played a toxic role in B. rapa under Se(IV) stress by triggering ROS burst. Such findings can be used to evaluate the toxic effects of Se contamination on crop plants. PMID:25333984

Chen, Yi; Mo, Hai-Zhen; Hu, Liang-Bin; Li, You-Qin; Chen, Jian; Yang, Li-Fei

2014-01-01

302

Exacerbated Transplant Arteriosclerosis in Inducible Nitric Oxide-Deficient Mice  

Microsoft Academic Search

Background—Inducible NO synthase (NOS2, or iNOS) is upregulated in grafts with transplant arteriosclerosis. However, the functional role of NOS2 in the pathogenesis of transplant arteriosclerosis remains unclear. NOS2 may regulate lesion development by modulating the early alloimmune response and\\/or late myointimal thickening. Methods and Results—To determine whether NOS2-mediated pathways protect against or promote transplant arterioscle- rosis, we used NOS2-deficient mice

Jorg Koglin; Troels Glysing-Jensen; John S. Mudgett; Mary E. Russell

2010-01-01

303

Immune-relevant thrombocytes of common carp undergo parasite-induced nitric oxide-mediated apoptosis.  

PubMed

Common carp thrombocytes account for 30-40% of peripheral blood leukocytes and are abundant in the healthy animals' spleen, the thrombopoietic organ. We show that, ex vivo, thrombocytes from healthy carp express a large number of immune-relevant genes, among which several cytokines and Toll-like receptors, clearly pointing at immune functions of carp thrombocytes. Few studies have described the role of fish thrombocytes during infection. Carp are natural host to two different but related protozoan parasites, Trypanoplasma borreli and Trypanosoma carassii, which reside in the blood and tissue fluids. We used the two parasites to undertake controlled studies on the role of fish thrombocytes during these infections. In vivo, but only during infection with T. borreli, thrombocytes were massively depleted from the blood and spleen leading to severe thrombocytopenia. Ex vivo, addition of nitric oxide induced a clear and rapid apoptosis of thrombocytes from healthy carp, supporting a role for nitric oxide-mediated control of immune-relevant thrombocytes during infection with T. borreli. The potential advantage for parasites to selectively deplete the host of thrombocytes via nitric oxide-induced apoptosis is discussed. PMID:25681740

Fink, Inge R; Ribeiro, Carla M S; Forlenza, Maria; Taverne-Thiele, Anja; Rombout, Jan H W M; Savelkoul, Huub F J; Wiegertjes, Geert F

2015-06-01

304

Influence of hypertension on nitric oxide synthase expression and vascular effects of lipopolysaccharide in rat mesenteric arteries  

PubMed Central

Experiments were designed to investigate the effects of the inducible nitric oxide synthase (iNOS) stimulator, lipopolysaccharide (LPS), on noradrenaline (NA) responses and on NOS activity and its expression in intact mesenteric resistance arteries (MRAs) from Wistar Kyoto (WKY) and spontaneously hypertensive (SHR) rats. In MRAs from WKY, LPS (10??g?ml?1; 1–5?h) reduced the vasoconstrictor responses to NA (0.1–30??M) in the presence, but not in the absence of L-arginine (L-Arg, 10??M). However, in SHR arteries, LPS induced an incubation-time dependent reduction of NA responses in the absence, as well as the presence, of L-Arg. The LPS inhibitory effect was reduced by the non-specific NOS inhibitor L-NG-nitroarginine methyl ester (L-NAME, 100??M) and the selective iNOS inhibitor, aminoguanidine (100??M). L-NAME alone similarly shifted the concentration-response curve to NA leftward in arteries from both strains, while aminoguanidine had no effect. L-Arg shifted the curve to NA rightward only in SHR MRAs. Basal activity of both iNOS and constitutive NOS (conversion of [3H]-L-Arg to [3H]-L-citrulline) was similar in arteries from both strains. After 5?h incubation with LPS, only iNOS activity in arteries from SHR was increased. Basal iNOS protein expression was undetectable; basal endothelial (eNOS) protein expression was similar in arteries from both strains, while neuronal (nNOS) was greater in arteries from SHR. LPS induced iNOS protein expression, that was higher in arteries from SHR than in those from WKY. These results indicate that NO production, via iNOS induction, is greater than in those from MRAs from SHR to WKY. PMID:10991910

Briones, Ana M; Alonso, María J; Marín, Jesús; Balfagón, Gloria; Salaices, Mercedes

2000-01-01

305

3,3'-Diindolylmethane inhibits lipopolysaccharide-induced microglial hyperactivation and attenuates brain inflammation.  

PubMed

Recent studies have revealed that microglial hyperactivation and neuroinflammation are implicated in development and progression of neurodegenerative diseases. In this study, we examined the beneficial effects of 3,3'-diindolylmethane (DIM) and indole-3-carbinol (I3C), dietary components found in cruciferous vegetables, on brain inflammation. DIM, a major metabolite of I3C, suppressed lipopolysaccharide (LPS)-induced expression of inducible nitric oxide synthase and cyclooxygenase-2 in BV-2 microglia, but I3C did not. DIM, but not I3C, attenuated DNA-binding activity of nuclear factor-?B (NF-?B) and phosphorylation of inhibitor of ?B, suggesting that DIM might inhibit microglial hyperactivation by attenuating inflammatory transcription factor NF-?B. In addition, DIM, but not I3C, protected primary cortical neurons from inflammatory toxicity induced by the conditioned media from LPS-stimulated BV-2 microglia, indicating that DIM might attenuate microglial hyperactivation-mediated neuronal death. In an in vivo model of neuroinflammation, DIM suppressed LPS-induced brain inflammation in mouse hippocampus, as determined by the number of Iba-1-positive cells and the mRNA expression of F4/80. Taken together, these results suggest that DIM may have beneficial potential against brain inflammation and neurodegenerative diseases through the negative regulation of the NF-?B signal pathway in microglia. PMID:24162184

Kim, Hyo Won; Kim, Jiyoung; Kim, Jaekyoon; Lee, Siyoung; Choi, Bo-Ryoung; Han, Jung-Soo; Lee, Ki Won; Lee, Hyong Joo

2014-01-01

306

Anti-inflammatory effect of Taraxacum officinale leaves on lipopolysaccharide-induced inflammatory responses in RAW 264.7 cells.  

PubMed

To investigate the efficacy and the mechanism of the anti-inflammatory effect of Taraxacum officinale leaves (TOLs), the effect of a methanol extract and its fractions recovered from TOLs on lipopolysaccharide (LPS)-induced responses was studied in the mouse macrophage cell line, RAW 264.7. Cells were pretreated with various concentrations of the methanol extract and its fractions and subsequently incubated with LPS (1 microg/mL). The levels of nitric oxide (NO), prostaglandin (PG) E(2), and pro-inflammatory cytokines including tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, and IL-6 were determined using enzyme-linked immunosorbent assays. Expressions of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 and activation of mitogen-activated protein (MAP) kinases were analyzed using western blotting. The methanol extract and its fractions inhibited LPS-induced production of NO, pro-inflammatory cytokines, and PGE(2) in a dose-dependent manner. The chloroform fraction significantly suppressed production of NO, PGE(2), and two pro-inflammatory cytokines (TNF-alpha and IL-1beta) in a dose-dependent manner with 50% inhibitory concentration values of 66.51, 90.96, 114.76, and 171.06 microg/mL, respectively. The ethyl acetate fraction also inhibited production of the inflammatory molecules. The chloroform and ethyl acetate fractions reduced LPS-induced expressions of iNOS and COX-2 and activation of MAP kinases in a dose-dependent manner. Among the fractions of the methanol extract, the chloroform and ethyl acetate fractions exhibited the most effective anti-inflammatory activities. These results show that the anti-inflammatory effects of TOLs are probably due to down-regulation of NO, PGE(2), and pro-inflammatory cytokines and reduced expressions of iNOS and COX-2 via inactivation of the MAP kinase signal pathway. PMID:20673058

Koh, Yoon-Jeoung; Cha, Dong-Soo; Ko, Je-Sang; Park, Hyun-Jin; Choi, Hee-Don

2010-08-01

307

Nitric oxide induces hydroxyl radical generation in rat hearts via depolarization-induced nitric oxide synthase activation  

Microsoft Academic Search

We examined the effect of NG-nitro-L-arginine methyl ester (L-NAME), a NOS inhibitor, on extracellular potassium ion concentration ([K+]o) and induced hydroxyl free radical (”OH) generation by an in vivo microdialysis technique. A flexibly mounted microdialysis technique was used to detect the generation of ”OH in in-vivo rat hearts. The microdialysis probe was implanted in the left ventricular myocardium of anesthetized

Toshio Obata; Yasumitsu Yamanaka

2001-01-01

308

Nitric oxide induces hydroxyl radical generation in rat hearts via depolarization-induced nitric oxide synthase activation.  

PubMed

We examined the effect of NG-nitro-L-arginine methyl ester (L-NAME), a NOS inhibitor, on extracellular potassium ion concentration ([K+]o) and induced hydroxyl free radical (.OH) generation by an in vivo microdialysis technique. A flexibly mounted microdialysis technique was used to detect the generation of .OH in in-vivo rat hearts. The microdialysis probe was implanted in the left ventricular myocardium of anesthetized rats and tissue was perfused with Ringer's solution through the microdialysis probe at a rate of 1.0 microl/min. To measure the level of .OH, sodium salicylate in Ringer's solution (0.5 nmol/microl per min) was infused directly through a microdialysis probe to detect the generation of .OH as reflected by the nonenzymatic formation of 2,3-dihydroxybenzoic acid (2,3-DHBA). Induction of high-concentration [K+]o (20, 70 and 140 mM) significantly increased formation of .OH trapped as 2,3-DHBA in a concentration-dependent manner. However, the application of L-NAME (50 mg/kg, i.v.) and allopurinol, a xanthine oxidase inhibitor, abolished the [K+]o depolarization-induced .OH generation. Tyramine (1.0 mM) increased the level of 2,3-DHBA. However, the application of L-NAME did not change the level of 2,3-DHBA. On the other hand, pretreatment with allopurinol (10 mg/kg, i.v.) abolished the KCl- or tyramine-induced .OH generation. Moreover, when iron (II) was administered to [K+]o (70 mM)-pretreated animals, there was a marked increased in the level of 2,3-DHBA. However, the application of L-NAME was not related to a Fenton-type reaction via [K+]o depolarization-induced .OH generation. To examine the effect of L-NAME on ischemic/reperfused rat myocardium, the heart was subjected to myocardial ischemia for 15 min by occlusion by left anterior descending coronary artery branch (LAD). When the heart was reperfused, a marked elevation of the level of 2,3-DHBA was observed. However, L-NAME attenuated .OH generation by ischemic/reperfused rat heart. These results suggest that NOS inhibition is associated with a cardioprotective effect due to the suppression of [K+]o depolarization-induced .OH generation. PMID:11485040

Obata, T; Yamanaka, Y

2001-07-01

309

Comparison of inducible nitric oxide synthase activity in pancreatic islets of young and aged rats  

PubMed Central

Objective(s): Some pathologic situations such as diabetes and metabolic syndrome are associated with alternation in nitric oxide level. Incidence of these condition increases with aging. On the other hand, insulin secretion is modulated by nitric oxide, and nitric oxide synthase (NOS) activity is also altered in diabetes. In this study, modification in the enzyme activity associated with aging and also optimized procedure for islet NOS assay was investigated. Materials and Methods: Male Wistar rats were randomly divided in two experimental groups: A: adult rats; were 4 month old and B: old rats; were 12 month old. In all groups, plasma glucose, insulin and NOX (nitrite + nitrate = NOX) were measured, and also insulin secretion in isolated pancreatic islet with or without L-NAME was investigated. Furthermore, the inducible NOS activity with L-citrulline measurement in islets was measured. Results: L-citrulline was quantified using one step HPLC column. Aging induced hyperglycemia (P<0.05) and excess plasma NOX (17.74 ± 1.664 and 26.25 ± 2.166 ?mol/l in A and B groups respectively, P<0.05) with unaltered plasma insulin. Islet insulin secretion was significantly reduced in aging rats. L-NAME induced islet insulin secretion especially in aging rats (P=0.003). Inducible NOS activity in islets of aging rats was significantly higher than adult rats (1.082 ± 0.084 and 6.277 ± 0.475 pmol/min per mg protein in adult and aging rats, respectively, P<0.001). Conclusion: These findings show that decreased in islet insulin secretion may be related to increase in iNOS activity in islets, which follows impaired carbohydrate metabolism in aging. PMID:25810884

Farrokhfall, Khadije; Hashtroudi, Mehri Seyed; Ghasemi, Asghar; Mehrani, Hossein

2015-01-01

310

Endogenous nitric oxide modifies antigen-induced microvascular leakage in sensitized guinea pig airways.  

PubMed

To examine the role of endogenous nitric oxide in allergic airway inflammation, we investigated the effect of a nitric oxide synthase inhibitor, N omega-nitro-L-arginine methyl ester (l-NAME), on antigen-induced airway microvascular leakage in actively sensitized guinea pigs by using Evans blue dye. Three weeks after sensitization with ovalbumin (10 micrograms), the tracheas were cannulated, and lungs were artificially ventilated. Animals were pretreated with atropine and propranolol (both 1 mg/kg, intravenously) to avoid neural modification. Ovalbumin inhalation (3 mg/ml, 1 minute) challenge caused significant microvascular leakage in all airways portions, which was significantly suppressed in a dose-dependent manner by pretreatment with intravenous injection of L-NAME (1 and 10 mg/kg) but not with the inactive enantiomer D-NAME (10 mg/kg). This inhibition by L-NAME was significantly reversed by co-administration of L-arginine (100 mg/kg, intravenously). Pretreatment with a vasoconstrictor, phenylephrine (20 micrograms/kg, intravenously), had no inhibitory effects on antigen-induced airway microvascular leakage despite increasing systemic blood pressure. Inhalation of representative mast cell-derived mediators, histamine (2 mg/ml, 1 minute) or leukotriene D4 (5 micrograms/ml, 1 minute), produced significant microvascular leakage in all airways. L-NAME (10 mg/kg, intravenously) partially but significantly inhibited leukotriene D4-induced leakage, whereas histamine-induced leakage was not affected. These results suggest that endogenous nitric oxide acts to increase airway microvascular leakage after airway allergic reaction. PMID:8765828

Miura, M; Ichinose, M; Kageyama, N; Tomaki, M; Takahashi, T; Ishikawa, J; Ohuchi, Y; Oyake, T; Endoh, N; Shirato, K

1996-07-01

311

Direct demonstration of insulin-like growth factor-I-induced nitric oxide production by endothelial cells  

Microsoft Academic Search

Direct demonstration of insulin-like growth factor-I-induced nitric oxide production by endothelial cells. Several lines of evidence indicate that insulin-like growth factor-I (IGF-I) is a potent mediator of vasodilation. To elucidate the mechanism and site of action of IGF-I, we performed continuous monitoring of nitric oxide (NO) release from endothelial cells using a highly-sensitive amperometric NO-sensor. Two types of cultured cells

Hirokazu Tsukahara; Dmitri V Gordienko; Burkhard Tonshoff; Marie C Gelato; Micheal S Goligorsky

1994-01-01

312

TNF-? dependent production of inducible nitric oxide is involved in PGE1 protection against acute liver injury  

Microsoft Academic Search

BACKGROUNDTumour necrosis factor ? (TNF-?) and nitric oxide modulate damage in several experimental models of liver injury. We have previously shown that protection against d-galactosamine (D-GalN) induced liver injury by prostaglandin E1 (PGE1) was accompanied by an increase in TNF-? and nitrite\\/nitrate in serum.AIMSThe aim of the present study was to evaluate the role of TNF-? and nitric oxide during

J Muntané; F J Rodríguez; O Segado; A Quintero; J M Lozano; E Siendones; C A Pedraza; M Delgado; F OValle; R García; J L Montero; M De la Mata; G Miño

2000-01-01

313

Different Aging Patterns of the Neuronal and Inducible Isoforms of Nitric Oxide Synthase in Mouse Hippocampus by Immunohistochemistry  

Microsoft Academic Search

Background: Nitric oxide synthase (NOS) produces nitric oxide (NO) from arginine, and both NOS and NO play important roles in the normal aging of the hippocampus. Aim: To study the age-related distribution patterns of the neuronal (nNOS) and the inducible (iNOS) isoforms of NOS in the mouse hippocampus, and their roles in the aging mechanism. Result: The nNOS significantly decreased

Ho Wai Wong; Ying Tat Mak; Mang Chung Yu; David Tai Wai Yew

2005-01-01

314

Nitric oxide-induced apoptotic cell death in the acute phase of Trypanosoma cruzi infection in mice  

Microsoft Academic Search

Production of gamma-interferon (IFN-?) and tumor necrosis factor-alpha (TNF-?) in Trypanosoma cruzi-infected mice results in the activation of inducible nitric oxide synthase (iNOS) and in elevated nitric oxide (NO) synthesis, which is important for the macrophage trypanocidal activity. However, NO has been shown to be involved in suppression of host immunity. In the present investigation, we studied the role of

Gislâine A Martins; Maria A. G Cardoso; Júlio C. S Aliberti; João S Silva

1998-01-01

315

Deficiency of neuronal nitric oxide synthase (nNOS) worsens alcohol-induced microencephaly and neuronal loss in developing mice  

Microsoft Academic Search

Previous work conducted in vitro suggests that nitric oxide (NO) protects developing neurons against the toxic effects of alcohol. We tested the hypothesis that neonatal mice carrying a null mutation for neuronal nitric oxide synthase (nNOS), the enzyme which synthesizes NO in neurons, have increased vulnerability to alcohol-induced microencephaly and neuronal loss. Wild-type mice and mutant (nNOS?\\/?) mice received a

Daniel J Bonthius; Georgios Tzouras; Bahri Karacay; Jolonda Mahoney; Ana Hutton; Ross McKim; Nicholas J Pantazis

2002-01-01

316

Nitric oxide: Mediator of nonadrenergic noncholinergic nerve-induced responses of opossum esophageal muscle  

SciTech Connect

Nonadrenergic noncholinergic (NANC) nerves of the opossum esophagus mediate relaxation of circular muscle from the lower esophageal sphincter (LES) and the off contraction of circular esophageal muscle. The latencies between the end of the stimulus and the off contraction describe a gradient such that the latency is longest in muscle from the caudad esophagus. N{sup G}-nitro-L-arginine (L-NNA), an inhibitor of nitric oxide synthase, and nitric oxide were used to test the hypothesis that NO is a mediator of these nerve-induced responses. Both electrical field stimulation (EFS) of intrinsic esophageal nerves and exogenous NO relaxed LES muscle. Only EFS-induced relaxation was inhibited by L-NNA. L-arginine, the substrate for NO synthase, antagonized the inhibitory effect of L-NNA. Exogenous NO neither relaxed nor contracted circular esophageal muscle. Both the amplitude and the latency of the off contraction were diminished by L-NNA. L-arginine antagonized the action of L-NNA. N{sup G}-nitro-L-arginine also attenuated the gradient in the latency of the off response by shortening latencies in muscle form the caudad esophagus. It had no effect on cholinergic nerve-induced contraction of longitudinal esophageal muscle. These data support the hypothesis that NO or an NO-containing compound mediates NANC nerve-induced responses of the esophagus and LES.

Murray, J.; Du, C.; Conklin, J.L.; Ledlow, A.; Bates, J.N. (Univ. of Iowa, Iowa City (United States))

1991-03-15

317

Mice lacking the gene for inducible or endothelial nitric oxide are resistant to sporocyst induced Sarcocystis neurona infections.  

PubMed

Equine protozoal myeloencephalitis (EPM) is a neurologic syndrome in horses from the Americas and is usually caused by infection with the apicomplexan parasite, Sarcocystis neurona. Little is known about the role of immunobiological mediators to this parasite. Nitric oxide (NO) is important in resistance to many intracellular parasites. We, therefore, investigated the role of inducible and endothelial NO in resistance to clinical disease caused by S. neurona in mice. Groups of interferon-gamma gene knockout (IFN-gamma-KO) mice, inducible nitric oxide synthase gene knockout (iNOS-KO) mice, endothelial nitric oxide synthase gene knockout (eNOS-KO) and appropriate genetic background mice (BALB/c or C57BL/6) were orally fed sporocysts or Hanks balanced salt solution. Mice were observed for signs of clinical disease and examined at necropsy. Clinical disease and deaths occurred only in the IFN-gamma-KO mice. Microscopic lesions were seen only in the brains of IFN-gamma-KO mice. Results of this study indicate that iNOS and eNOS are not major mediators of resistance to S. neurona infections. Results of this study suggest that IFN-gamma mediated immunity to S. neurona may be mediated by non-NO-dependent mechanisms. PMID:11777610

Rosypal, Alexa C; Lindsay, David S; Duncan, Robert; Ahmed, S Ansar; Zajac, Anne M; Dubey, J P

2002-02-01

318

Elicitor-induced nitric oxide burst is essential for triggering catharanthine synthesis in Catharanthus roseus suspension cells  

Microsoft Academic Search

Elicitor prepared from the cell walls of Penicillium citrinum induced multiple responses in Catharanthus roseus suspension cells, including rapid generation of nitric oxide (NO), sequentially followed by enhancement of catharanthine production by C. roseus cells. Elicitor-induced catharanthine biosynthesis was blocked by NO-specific scavenger 2-4-carboxyphenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide and nitric oxide synthase (NOS) inhibitor S,S'-1,3-phenylene-bis(1,2-ethanediyl)-bis-isothiourea (PBITU). PBITU also strongly inhibited elicitor-induced NO generation by

Maojun Xu; Jufang Dong

2005-01-01

319

Rabdosia japonica var. glaucocalyx Flavonoids Fraction Attenuates Lipopolysaccharide-Induced Acute Lung Injury in Mice  

PubMed Central

Rabdosia japonica var. glaucocalyx (Maxim.) Hara, belonging to the Labiatae family, is widely used as an anti-inflammatory and antitumor drug for the treatment of different inflammations and cancers. Aim of the Study. To investigate therapeutic effects and possible mechanism of the flavonoids fraction of Rabdosia japonica var. glaucocalyx (Maxim.) Hara (RJFs) in acute lung injury (ALI) mice induced by lipopolysaccharide (LPS). Materials and Methods. Mice were orally administrated with RJFs (6.4, 12.8, and 25.6?mg/kg) per day for 7 days, consecutively, before LPS challenge. Lung specimens and the bronchoalveolar lavage fluid (BALF) were isolated for histopathological examinations and biochemical analysis. The level of complement 3 (C3) in serum was quantified by a sandwich ELISA kit. Results. RJFs significantly attenuated LPS-induced ALI via reducing productions of the level of inflammatory mediators (TNF-?, IL-6, and IL-1?), and significantly reduced complement deposition with decreasing the level of C3 in serum, which was exhibited together with the lowered myeloperoxidase (MPO) activity and nitric oxide (NO) and protein concentration in BALF. Conclusions. RJFs significantly attenuate LPS-induced ALI via reducing productions of proinflammatory mediators, decreasing the level of complement, and reducing radicals. PMID:25013450

Xu, Nai-yu; Li, Xian-lun; Xia, Long; Zhang, Jian; Liang, Zhi-tao; Zhao, Zhong-zhen; Chen, Dao-feng

2014-01-01

320

Hydrogen sulfide inhibits nitric oxide production and nuclear factor-kappaB via heme oxygenase-1 expression in RAW264.7 macrophages stimulated with lipopolysaccharide.  

PubMed

Hydrogen sulfide (H(2)S), a regulatory gaseous molecule that is endogenously synthesized by cystathionine gamma-lyase (CSE) and/or cystathionine beta-synthase (CBS) from L-cysteine (L-Cys) metabolism, is a putative vasodilator, and its role in nitric oxide (NO) production is unexplored. Here, we show that at noncytotoxic concentrations, H(2)S was able to inhibit NO production and inducible NO synthase (iNOS) expression via heme oxygenase (HO-1) expression in RAW264.7 macrophages stimulated with lipopolysaccharide (LPS). Both H(2)S solution prepared by bubbling pure H(2)S gas and NaSH, a H(2)S donor, dose dependently induced HO-1 expression through the activation of the extracellular signal-regulated kinase (ERK). Pretreatment with H(2)S or NaHS significantly inhibited LPS-induced iNOS expression and NO production. Moreover, NO production in LPS-stimulated macrophages that are expressing CSE mRNA was significantly reduced by the addition of L-Cys, a substrate for H(2)S, but enhanced by the selective CSE inhibitor beta-cyano-L-alanine but not by the CBS inhibitor aminooxyacetic acid. While either blockage of HO activity by the HO inhibitor, tin protoporphyrin IX, or down-regulation of HO-1 expression by HO-1 small interfering RNA (siRNA) reversed the inhibitory effects of H(2)S on iNOS expression and NO production, HO-1 overexpression produced the same inhibitory effects of H(2)S. In addition, LPS-induced nuclear factor (NF)-kappaB activation was diminished in RAW264.7 macrophages preincubated with H(2)S. Interestingly, the inhibitory effect of H(2)S on NF-kappaB activation was reversed by the transient transfection with HO-1 siRNA, but was mimicked by either HO-1 gene transfection or treatment with carbon monoxide (CO), an end product of HO-1. CO treatment also inhibited LPS-induced NO production and iNOS expression via its inactivation of NF-kappaB. Collectively, our results suggest that H(2)S can inhibit NO production and NF-kappaB activation in LPS-stimulated macrophages through a mechanism that involves the action of HO-1/CO. PMID:16781459

Oh, Gi-Su; Pae, Hyun-Ock; Lee, Bok-Soo; Kim, Byeong-Nam; Kim, Jong-Moon; Kim, Hyung-Ryong; Jeon, Seon Bok; Jeon, Woo Kyu; Chae, Han-Jung; Chung, Hun-Taeg

2006-07-01

321

The inhibition of lipopolysaccharide-induced macrophage inflammation by 4 compounds in Hypericum perforatum extract is partially dependent on the activation of SOCS3  

PubMed Central

Our previous studies found that 4 compounds, namely pseudohypericin, amentoflavone, quercetin, and chlorogenic acid in Hypericum perforatum ethanol extract synergistically inhibited lipopolysaccharide (LPS)-induced macrophage production of prostaglandin E2 (PGE2). Microarray studies led us to hypothesize that these compounds inhibited PGE2 production by activating suppressor of cytokine signaling 3 (SOCS3). In the current study we used siRNA to knockdown the expression of SOCS3 in RAW 264.7 macrophages and investigated the impact of H. perforatum extract and the 4 compounds on inflammatory mediators and cytokines. We found SOCS3 knockdown significantly compromised the inhibition of PGE2 and nitric oxide (NO) by the 4 compounds, but not by the extract. The 4 compounds, but not the extract decreased interleukin-6 (IL-6) and tumor necrosis factor-? (TNF-?), while both of them lowered interleukine-1?. SOCS3 knockdown further decreased IL-6 and TNF-?. Pseudohypericin was the major contributor to the PGE2 and NO inhibition in cells treated with the 4 compounds and its activity was lost with SOCS3 knockdown. Cyclooxygenase-2 (COX-2) and inducible NO synthase protein expression were not altered by the treatments, while COX-2 activity was decreased by the extract and the 4 compounds and increased by SOCS3 knockdown. In summary, we demonstrated that the 4 compounds inhibited LPS-induced PGE2 and NO through SOCS3 activation. The reduction of PGE2 can be partially attributed to COX-2 enzyme activity, which was significantly elevated with SOCS3 knockdown. At the same time, our results also suggest that constituents in H. perforatum extract were alleviating LPS-induced macrophage response through SOCS3 independent mechanisms. PMID:22245632

Huang, Nan; Rizshsky, Ludmila; Hauck, Catherine C.; Nikolau, Basil J.; Murphy, Patricia A.; Birt, Diane F.

2012-01-01

322

Foeniculum vulgare Mill. Protects against Lipopolysaccharide-induced Acute Lung Injury in Mice through ERK-dependent NF-?B Activation.  

PubMed

Foeniculum vulgare Mill. (fennel) is used to flavor food, in cosmetics, as an antioxidant, and to treat microbial, diabetic and common inflammation. No study to date, however, has assessed the anti-inflammatory effects of fennel in experimental models of inflammation. The aims of this study were to investigate the anti-inflammatory effects of fennel in model of lipopolysaccharide (LPS)-induced acute lung injury. Mice were randomly assigned to seven groups (n=7~10). In five groups, the mice were intraperitoneally injected with 1% Tween 80-saline (vehicle), fennel (125, 250, 500µl/kg), or dexamethasone (1 mg/kg), followed 1 h later by intratracheal instillation of LPS (1.5 mg/kg). In two groups, the mice were intraperitoneally injected with vehicle or fennel (250µl/kg), followed 1 h later by intratracheal instillation of sterile saline. Mice were sacrificed 4 h later, and bronchoalveolar lavage fluid (BALF) and lung tissues were obtained. Fennel significantly and dose-dependently reduced LDH activity and immune cell numbers in LPS treated mice. In addition fennel effectively suppressed the LPS-induced increases in the production of the inflammatory cytokines interleukin-6 and tumor necrosis factor-alpha, with 500µl/kg fennel showing maximal reduction. Fennel also significantly and dose-dependently reduced the activity of the proinflammatory mediator matrix metalloproteinase 9 and the immune modulator nitric oxide (NO). Assessments of the involvement of the MAPK signaling pathway showed that fennel significantly decreased the LPS-induced phosphorylation of ERK. Fennel effectively blocked the inflammatory processes induced by LPS, by regulating pro-inflammatory cytokine production, transcription factors, and NO. PMID:25729281

Lee, Hui Su; Kang, Purum; Kim, Ka Young; Seol, Geun Hee

2015-03-01

323

Foeniculum vulgare Mill. Protects against Lipopolysaccharide-induced Acute Lung Injury in Mice through ERK-dependent NF-?B Activation  

PubMed Central

Foeniculum vulgare Mill. (fennel) is used to flavor food, in cosmetics, as an antioxidant, and to treat microbial, diabetic and common inflammation. No study to date, however, has assessed the anti-inflammatory effects of fennel in experimental models of inflammation. The aims of this study were to investigate the anti-inflammatory effects of fennel in model of lipopolysaccharide (LPS)-induced acute lung injury. Mice were randomly assigned to seven groups (n=7~10). In five groups, the mice were intraperitoneally injected with 1% Tween 80-saline (vehicle), fennel (125, 250, 500µl/kg), or dexamethasone (1 mg/kg), followed 1 h later by intratracheal instillation of LPS (1.5 mg/kg). In two groups, the mice were intraperitoneally injected with vehicle or fennel (250µl/kg), followed 1 h later by intratracheal instillation of sterile saline. Mice were sacrificed 4 h later, and bronchoalveolar lavage fluid (BALF) and lung tissues were obtained. Fennel significantly and dose-dependently reduced LDH activity and immune cell numbers in LPS treated mice. In addition fennel effectively suppressed the LPS-induced increases in the production of the inflammatory cytokines interleukin-6 and tumor necrosis factor-alpha, with 500µl/kg fennel showing maximal reduction. Fennel also significantly and dose-dependently reduced the activity of the proinflammatory mediator matrix metalloproteinase 9 and the immune modulator nitric oxide (NO). Assessments of the involvement of the MAPK signaling pathway showed that fennel significantly decreased the LPS-induced phosphorylation of ERK. Fennel effectively blocked the inflammatory processes induced by LPS, by regulating pro-inflammatory cytokine production, transcription factors, and NO. PMID:25729281

Lee, Hui Su; Kang, Purum; Kim, Ka Young

2015-01-01

324

Forsythiaside attenuates lipopolysaccharide-induced inflammatory responses in the bursa of Fabricius of chickens by downregulating the NF-?B signaling pathway  

PubMed Central

Forsythiaside, a phenylethanoside product isolated from air-dried fruits of Forsythia suspensa, has been demonstrated to exhibit antioxidant, antibacterial and anti-inflammatory activities in vitro. However, its mechanism and the effects of lipopolysaccharide (LPS)-induced injury on the bursa of Fabricius (BF) of chickens are poorly understood. The present study aimed to investigate the anti-inflammatory effects of forsythiaside on LPS-induced acute inflammation. In addition, the potential molecular mechanisms of forsythiaside were analyzed in the BF, a special immune organ in chickens. Forty 15-day-old chickens were randomly divided into control, LPS and LPS plus forsythiaside (30 or 60 mg/kg) groups (n=10 for each group). In the LPS plus forsythiaside (30 or 60 mg/kg) groups, the chickens were orally administered with forsythiaside at doses of 30 and 60 mg/kg for seven days. At 21 days old, the chickens were intravenously injected with 200 ?g/kg body weight LPS. Chickens in the control and LPS groups were only administered with vehicle or LPS, respectively, at day 21. At 3 h post-injection, the body temperature and nitric oxide (NO) levels were analyzed. In addition, the levels and mRNA expression of pro-inflammatory cytokines, including tumor necrosis factor-? (TNF-?), interleukin-6 (IL-6) and IL-1?, and the mRNA expression of nuclear factor-?B (NF-?B), cyclooxygenase-2 (COX-2) and inducible NO synthase (iNOS), were examined in the BFs isolated from the chickens. The results revealed that forsythiaside was able to attenuate the LPS-induced inflammatory responses in the BFs of the chickens. The mechanisms by which forsythiaside exerted its anti-inflammatory effect were found to correlate with the inhibition of IL-6, IL-1?, TNF-? and COX-2 production, via the inactivation of NF-?B, indicating that the NF-?B-iNOS-NO signaling pathway may be important in this process. PMID:24348786

CHENG, GUANGDONG; ZHAO, YULIAN; LI, HE; WU, YUE; LI, XIANXIAN; HAN, QIANG; DAI, CHONGSHAN; LI, YANHUA

2014-01-01

325

Evidence of nitric-oxide-induced surface band bending of indium tin oxide  

NASA Astrophysics Data System (ADS)

The interaction of indium tin oxide (ITO) film with nitric oxide (NO) has been investigated in situ by a four-point probe and x-ray photoelectron spectroscopy (XPS). The XPS N 1s peak emerged at a high binding energy of 404 eV indicating that NO was molecularly adsorbed on ITO surface. The adsorption of NO on ITO surface also induced a 0.2 eV shift in its valence band maximum to the low binding energy side leading to an upward surface band bending. We have shown that the increase in the ITO sheet resistance was attributed to its surface band bending.

Hu, Jianqiao; Pan, Jisheng; Zhu, Furong; Gong, Hao

2004-06-01

326

Arsenic toxicity induced endothelial dysfunction and dementia: Pharmacological interdiction by histone deacetylase and inducible nitric oxide synthase inhibitors  

SciTech Connect

Arsenic toxicity has been reported to damage all the major organs including the brain and vasculature. Dementia including Alzheimer's disease (AD) and vascular dementia (VaD) are posing greater risk to the world population as it is now increasing at a faster rate. We have investigated the role of sodium butyrate, a selective histone deacetylase (HDAC) inhibitor and aminoguanidine, a selective inducible nitric oxide synthase (iNOS) inhibitor in pharmacological interdiction of arsenic toxicity induced vascular endothelial dysfunction and dementia in rats. Arsenic toxicity was done by administering arsenic drinking water to rats. Morris water-maze (MWM) test was used for assessment of learning and memory. Endothelial function was assessed using student physiograph. Oxidative stress (aortic superoxide anion, serum and brain thiobarbituric acid reactive species, brain glutathione) and nitric oxide levels (serum nitrite/nitrate) were also measured. Arsenic treated rats have shown impairment of endothelial function, learning and memory, reduction in serum nitrite/nitrate and brain GSH levels along with increase in serum and brain TBARS. Sodium butyrate as well as aminoguanidine significantly convalesce arsenic induced impairment of learning, memory, endothelial function, and alterations in various biochemical parameters. It may be concluded that arsenic induces endothelial dysfunction and dementia, whereas, sodium butyrate, a HDAC inhibitor as well as aminoguanidine, a selective iNOS inhibitor may be considered as potential agents for the management of arsenic induced endothelial dysfunction and dementia. - Highlights: • As has induced endothelial dysfunction (Edf) and vascular dementia (VaD). • As has increased oxidative stress, AChE activity and decreased serum NO. • Inhibitors of HDAC and iNOS have attenuated As induced Edf and VaD. • Both the inhibitors have attenuated As induced biochemical changes. • Inhibitor of HDAC and iNOS has shown good potential in As induced VaD.

Sharma, Bhupesh, E-mail: drbhupeshresearch@gmail.com; Sharma, P.M.

2013-11-15

327

Role of nitric oxide in adenosine-induced vasodilation in humans  

NASA Technical Reports Server (NTRS)

Vasodilation is one of the most prominent effects of adenosine and one of the first to be recognized, but its mechanism of action is not completely understood. In particular, there is conflicting information about the potential contribution of endothelial factors. The purpose of this study was to explore the role of nitric oxide in the vasodilatory effect of adenosine. Forearm blood flow responses to intrabrachial adenosine infusion (125 microg/min) were assessed with venous occlusion plethysmography during intrabrachial infusion of saline or the nitric oxide synthase inhibitor NG-monomethyl-L-arginine (L-NMMA) (12.5 mg/min). Intrabrachial infusions of acetylcholine (50 microg/min) and nitroprusside (3 microg/min) were used as a positive and negative control, respectively. These doses were chosen to produce comparable levels of vasodilation. In a separate study, a second saline infusion was administered instead of L-NMMA to rule out time-related effects. As expected, pretreatment with L-NMMA reduced acetylcholine-induced vasodilation; 50 microg/min acetylcholine increased forearm blood flow by 150+/-43% and 51+/-12% during saline and L-NMMA infusion, respectively (P<.01, n=6). In contrast, L-NMMA did not affect the increase in forearm blood flow produced by 3 microg/min nitroprusside (165+/-30% and 248+/-41% during saline and L-NMMA, respectively) or adenosine (173+/-48% and 270+/-75% during saline and L-NMMA, respectively). On the basis of our observations, we conclude that adenosine-induced vasodilation is not mediated by nitric oxide in the human forearm.

Costa, F.; Biaggioni, I.; Robertson, D. (Principal Investigator)

1998-01-01

328

Nitric Oxide Acts as a Positive Regulator to Induce Metamorphosis of the Ascidian Herdmania momus  

PubMed Central

Marine invertebrates commonly have a biphasic life cycle in which the metamorphic transition from a pelagic larva to a benthic post-larva is mediated by the nitric oxide signalling pathway. Nitric oxide (NO) is synthesised by nitric oxide synthase (NOS), which is a client protein of the molecular chaperon heat shock protein 90 (HSP90). It is notable, then, that both NO and HSP90 have been implicated in regulating metamorphosis in marine invertebrates as diverse as urochordates, echinoderms, molluscs, annelids, and crustaceans. Specifically, the suppression of NOS activity by the application of either NOS- or HSP90-inhibiting pharmacological agents has been shown consistently to induce the initiation of metamorphosis, leading to the hypothesis that a negative regulatory role of NO is widely conserved in biphasic life cycles. Further, the induction of metamorphosis by heat-shock has been demonstrated for multiple species. Here, we investigate the regulatory role of NO in induction of metamorphosis of the solitary tropical ascidian, Herdmania momus. By coupling pharmacological treatments with analysis of HmNOS and HmHSP90 gene expression, we present compelling evidence of a positive regulatory role for NO in metamorphosis of this species, in contrast to all existing ascidian data that supports the hypothesis of NO as a conserved negative regulator of metamorphosis. The exposure of competent H. momus larvae to a NOS inhibitor or an NO donor results in an up-regulation of NOS and HSP90 genes. Heat shock of competent larvae induces metamorphosis in a temperature dependent manner, up to a thermal tolerance that approaches 35°C. Both larval/post-larval survival and the appearance of abnormal morphologies in H. momus post-larvae reflect the magnitude of up-regulation of the HSP90 gene in response to heat-shock. The demonstrated role of NO as a positive metamorphic regulator in H. momus suggests the existence of inter-specific adaptations of NO regulation in ascidian metamorphosis. PMID:24019877

Ueda, Nobuo; Degnan, Sandie M.

2013-01-01

329

Morphine sensitization in the pentylenetetrazole-induced clonic seizure threshold in mice: role of nitric oxide and ? receptors.  

PubMed

Behavioral sensitization occurs after repeated administration of ?-opioid receptor agonists following a drug-free period. It seems that the changes in dopaminergic systems induced by ?-opioid receptor agonists play a crucial role in behavioral sensitization to opioids. Nitric oxide also plays a role in some behavioral effects of morphine, including sensitization to the locomotor-stimulating effect. This study investigated whether morphine sensitization appears in seizure threshold and the possible role of ?-opioid receptor and nitric oxide in this sensitization. Sensitization was produced by daily injections of morphine (0.1, 0.5, 1, 5, 15, or 30 mg/kg), followed by a 10-day washout period. Then the challenge test was performed using morphine (0.1, 0.5, 1, 5, 15, or 30 mg/kg) in different groups. To assess clonic seizure threshold, pentylenetetrazole (PTZ) was administered intravenously. Subcutaneous administration of morphine (0.1 and 0.5 mg/kg) induced sensitization in PTZ-induced clonic seizures in mice. Intraperitoneal administration of L-NAME (20 mg/kg), a nonselective inhibitor of nitric oxide synthase, or naltrexone (10 mg/kg), an opioid receptor antagonist, along with morphine inhibited morphine-induced sensitization in PTZ-induced seizure threshold. In conclusion, at low doses, morphine induces sensitization in PTZ-induced clonic seizures in mice probably as a result of the interaction with ?-receptors and nitric oxide. PMID:21419715

Shafaroodi, Hamed; Baradaran, Nazanin; Moezi, Leila; Dehpour, Siavash; Kabiri, Tina; Dehpour, Ahmad R

2011-04-01

330

Radiation-induced nitric oxide mitigates tumor hypoxia and radioresistance in a murine SCCVII tumor model  

SciTech Connect

Highlights: •IR-induced NO increased tissue perfusion and pO{sub 2}. •IR increased NO production in tumors without changes in the mRNA and protein levels of NOS isoforms. •NOS activity assay showed that IR upregulated eNOS activity in tumors. •IR-induced NO decreased tumor hypoxia and altered tumor radiosensitivity. -- Abstract: Tumor hypoxia, which occurs mainly as a result of inadequate tissue perfusion in solid tumors, is a well-known challenge for successful radiotherapy. Recent evidence suggests that ionizing radiation (IR) upregulates nitric oxide (NO) production and that IR-induced NO has the potential to increase intratumoral circulation. However, the kinetics of NO production and the responsible isoforms for NO synthase in tumors exposed to IR remain unclear. In this study, we aimed to elucidate the mechanism by which IR stimulates NO production in tumors and the effect of IR-induced NO on tumor radiosensitivity. Hoechst33342 perfusion assay and electron spin resonance oxymetry showed that IR increased tissue perfusion and pO{sub 2} in tumor tissue. Immunohistochemical analysis using two different hypoxic probes showed that IR decreased hypoxic regions in tumors; treatment with a nitric oxide synthase (NOS) inhibitor, L-NAME, abrogated the effects of IR. Moreover, IR increased endothelial NOS (eNOS) activity without affecting its mRNA or protein expression levels in SCCVII-transplanted tumors. Tumor growth delay assay showed that L-NAME decreased the anti-tumor effect of fractionated radiation (10 Gy × 2). These results suggested that IR increased eNOS activity and subsequent tissue perfusion in tumors. Increases in intratumoral circulation simultaneously decreased tumor hypoxia. As a result, IR-induced NO increased tumor radiosensitivity. Our study provides a new insight into the NO-dependent mechanism for efficient fractionated radiotherapy.

Nagane, Masaki, E-mail: nagane@vetmed.hokudai.ac.jp [Laboratory of Radiation Biology, Department of Environmental Veterinary Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo (Japan)] [Laboratory of Radiation Biology, Department of Environmental Veterinary Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo (Japan); Yasui, Hironobu, E-mail: yassan@vetmed.hokudai.ac.jp [Laboratory of Radiation Biology, Department of Environmental Veterinary Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo (Japan)] [Laboratory of Radiation Biology, Department of Environmental Veterinary Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo (Japan); Yamamori, Tohru, E-mail: yamamorit@vetmed.hokudai.ac.jp [Laboratory of Radiation Biology, Department of Environmental Veterinary Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo (Japan)] [Laboratory of Radiation Biology, Department of Environmental Veterinary Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo (Japan); Zhao, Songji, E-mail: zsi@med.hokudai.ac.jp [Department of Tracer Kinetics and Bioanalysis, Graduate School of Medicine, Hokkaido University, Sapporo (Japan)] [Department of Tracer Kinetics and Bioanalysis, Graduate School of Medicine, Hokkaido University, Sapporo (Japan); Kuge, Yuji, E-mail: kuge@med.hokudai.ac.jp [Central Institute of Isotope Science, Hokkaido University, Sapporo (Japan)] [Central Institute of Isotope Science, Hokkaido University, Sapporo (Japan); Tamaki, Nagara, E-mail: natamaki@med.hokudai.ac.jp [Department of Nuclear Medicine, Graduate School of Medicine, Hokkaido University, Sapporo (Japan)] [Department of Nuclear Medicine, Graduate School of Medicine, Hokkaido University, Sapporo (Japan); Kameya, Hiromi, E-mail: kameya@affrc.go.jp [Food Safety Division, National Food Research Institute, Tsukuba (Japan)] [Food Safety Division, National Food Research Institute, Tsukuba (Japan); Nakamura, Hideo, E-mail: naka@science-edu.org [Department of Chemistry, Hokkaido University of Education, Hakodate (Japan)] [Department of Chemistry, Hokkaido University of Education, Hakodate (Japan); Fujii, Hirotada, E-mail: hgfujii@sapmed.ac.jp [Center for Medical Education, Sapporo Medical University, Sapporo (Japan)] [Center for Medical Education, Sapporo Medical University, Sapporo (Japan); Inanami, Osamu, E-mail: inanami@vetmed.hokudai.ac.jp [Laboratory of Radiation Biology, Department of Environmental Veterinary Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo (Japan)] [Laboratory of Radiation Biology, Department of Environmental Veterinary Sciences, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo (Japan)

2013-08-02

331

Inhibition of inducible nitric oxide synthase and osteoclastic differentiation by Atractylodis Rhizoma Alba extract  

PubMed Central

Background: Atractylodis Rhizoma Alba (ARA) has been used in Korean folk medicine for constipation, dizziness, and anticancer agent. In the present study, we performed to test whether the methanolic extract of ARA has antioxidant and antiosteoclastogenesis activity in RAW 264.7 macrophage cells. Materials and Methods: Antioxidant capacities were tested by measuring free radical scavenging activity, nitric oxide (NO) levels, reducing power, and inducible nitric oxide synthase (iNOS) expression in response to lipopolysaccharides (LPS). Antiosteoclastogenesis activity was evaluated by performing tartrate-resistant acid phosphatase assay in RAW 264.7 macrophage cells. Results: The extract exerted significant 1,1-diphenyl-2-picrylhydrazyl and NO radical scavenging activity, and it exerted dramatic reducing power. Induction of iNOS and NO by LPS in RAW 264.7 cells was significantly inhibited by the extract, suggesting that the ARA extract inhibits NO production by suppressing iNOS expression. Strikingly, the ARA extracts substantially inhibited the receptor activator of NF-?B ligand-induced osteclastic differentiation of LPS-activated RAW 264.7 cells. The ARA extract contains a significant amount of antioxidant components, including phenolics, flavonoids and anthocyanins. Conclusion: These results suggest that the methanolic extract of ARA exerts significant antioxidant activities potentially via inhibiting free radicals and iNOS induction, thereby leading to the inhibition of osteoclastogenesis. PMID:25298665

Choi, Sung-Ho; Kim, Sung-Jin

2014-01-01

332

Inhibitory effects of 4-hydroxyderricin and xanthoangelol on lipopolysaccharide-induced inflammatory responses in RAW264 macrophages.  

PubMed

The Japanese herb, Ashitaba (Angelica keiskei Koidzumi), contains two prenylated chalcones, 4-hydroxyderricin and xanthoangelol, which are considered to be the major active compounds of Ashitaba. However, their effects on inflammatory responses are poorly understood. In the present study, we investigated the effects and underlying molecular mechanisms of 4-hydroxyderricin and xanthoangelol on lipopolysaccharide (LPS)-induced inflammatory responses in RAW264 mouse macrophages. LPS-mediated production of nitric oxide (NO) was markedly reduced by 4-hydroxyderricin (10 ?M) and xanthoangelol (5 ?M) compared with their parent compound, chalcone (25 ?M). They also inhibited LPS-induced secretion of tumor necrosis factor-alpha (TNF-?) and expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2). Although chalcone decreased the DNA-binding activity of both activator protein-1 (AP-1) and nuclear factor-kappa B (NF-?B), 4-hydroxyderricin and xanthoangelol suppressed only AP-1 and had no effect on NF-?B. On the other hand, all of the tested chalcones reduced the phosphorylation (at serine 536) level of the p65 subunit of NF-?B. 4-Hydroxyderricin and xanthoangelol may be promising for the prevention of inflammatory diseases. PMID:24369884

Yasuda, Michiko; Kawabata, Kyuichi; Miyashita, Miki; Okumura, Mayu; Yamamoto, Norio; Takahashi, Masakazu; Ashida, Hitoshi; Ohigashi, Hajime

2014-01-15

333

Sargachromenol from Sargassum micracanthum Inhibits the Lipopolysaccharide-Induced Production of Inflammatory Mediators in RAW 264.7 Macrophages  

PubMed Central

During our ongoing screening program designed to determine the anti-inflammatory potential of natural compounds, we isolated sargachromenol from Sargassum micracanthum. In the present study, we investigated the anti-inflammatory effects of sargachromenol on lipopolysaccharide (LPS)-induced inflammation in murine RAW 264.7 macrophage cells and the underlying mechanisms. Sargachromenol significantly inhibited the LPS-induced production of nitric oxide (NO) and prostaglandin E2 (PGE2) in a dose-dependent manner. It also significantly inhibited the protein expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) in a dose-dependent manner in LPS-stimulated macrophage cells. Further analyses showed that sargachromenol decreased the cytoplasmic loss of inhibitor ?B? (I?B?) protein. These results suggest that sargachromenol may exert its anti-inflammatory effects on LPS-stimulated macrophage cells by inhibiting the activation of the NF-?B signaling pathway. In conclusion, to our knowledge, this is the first study to show that sargachromenol isolated from S. micracanthum has an effective anti-inflammatory activity. Therefore, sargachromenol might be useful for cosmetic, food, or medical applications requiring anti-inflammatory properties. PMID:24194688

Yang, Eun-Jin; Ham, Young Min; Yang, Kyong-Wol; Lee, Nam Ho

2013-01-01

334

Nitric oxide enhances MPP +-induced hydroxyl radical generation via depolarization activated nitric oxide synthase in rat striatum  

Microsoft Academic Search

We examined the effect of NG-nitro-l-arginine methyl ester (l-NAME), a nitric oxide synthase (NOS) inhibitor, on extracellular potassium ion concentration ([K+]o)-enhanced hydroxyl radical (·OH) generation due to 1-methyl-4-phenylpyridinium ion (MPP+) was examined in the rat striatum. Rats were anesthetized, and sodium salicylate in Ringer’s solution (0.5 nmol\\/?l per min) was infused through a microdialysis probe to detect the generation of

Toshio Obata; Yasumitsu Yamanaka

2001-01-01

335

Bursopentin (BP5) Protects Dendritic Cells from Lipopolysaccharide-Induced Oxidative Stress for Immunosuppression  

PubMed Central

Dendritic cells (DCs) play a vital role in the regulation of immune-mediated inflammatory diseases. Thus, DCs have been regarded as a major target for the development of immunomodulators. However, oxidative stress could disturb inflammatory regulation in DCs. Here, we examined the effect of bursopentine (BP5), a novel pentapeptide isolated from chicken bursa of fabricius, on the protection of DCs against oxidative stress for immunosuppression. BP5 showed potent protective effects against the lipopolysaccharide (LPS)-induced oxidative stress in DCs, including nitric oxide, reactive oxygen species and lipid peroxidation. Furthermore, BP5 elevated the level of cellular reductive status through increasing the reduced glutathione (GSH) and the GSH/GSSG ratio. Concomitant with these, the activities of several antioxidative redox enzymes, including glutathione peroxidase (GPx), catalase (CAT) and superoxide dismutase (SOD), were obviously enhanced. BP5 also suppressed submucosal DC maturation in the LPS-stimulated intestinal epithelial cells (ECs)/DCs coculture system. Finally, we found that heme oxygenase 1 (HO-1) was remarkably upregulated by BP5 in the LPS-induced DCs, and played an important role in the suppression of oxidative stress and DC maturation. These results suggested that BP5 could protect the LPS-activated DCs against oxidative stress and have potential applications in DC-related inflammatory responses. PMID:25659113

Qin, Tao; Yin, Yinyan; Yu, Qinghua; Yang, Qian

2015-01-01

336

Beneficial Effects of Concomitant Neuronal and Inducible Nitric Oxide Synthase Inhibition in Ovine Burn and Inhalation Injury  

PubMed Central

Different isoforms of nitric oxide synthase are critically involved in the development of pulmonary failure secondary to acute lung injury. Here we tested the hypothesis that simultaneous blockade of inducible and neuronal nitric oxide synthase effectively prevents the pulmonary lesions in an ovine model of acute respiratory distress syndrome (ARDS) induced by combined burn and smoke inhalation injury. Chronically instrumented sheep were allocated to a sham-injured group (n = 6), an injured and untreated group (n = 6), or an injured group treated with simultaneous infusion of selective inducible and neuronal nitric oxide synthase inhibitors (n = 5). The injury was induced by 48 breath of cotton smoke and a 3rd degree burn of 40% total body surface area. All sheep were mechanically ventilated and fluid resuscitated. The injury induced severe pulmonary dysfunction as indicated by decreases in PaO2/FiO2 ratio and increases in pulmonary shunt fraction, ventilatory pressures, lung lymph flow, and lung wet/dry weight ratio. The treatment fully prevented the elevations in lymph and plasma nitrate/nitrite levels, pulmonary shunting, ventilatory pressures, lung lymph flow, and wet/dry weight ratio and significantly attenuated the decline in PaO2/FiO2 ratio. In conclusion, simultaneous blockade of inducible and neuronal nitric oxide synthase exerts beneficial pulmonary effects in an ovine model of ARDS secondary to combined burn and smoke inhalation injury. This novel treatment strategy may represent a useful therapeutic adjunct for patients with these injuries. PMID:21263377

Lange, Matthias; Hamahata, Atsumori; Enkhbaatar, Perenlei; Cox, Robert A.; Nakano, Yoshimitsu; Westphal, Martin; Traber, Lillian D.; Herndon, David N.; Traber, Daniel L.

2013-01-01

337

Tumor necrosis factor alpha acts as an autocrine second signal with gamma interferon to induce nitric oxide in group B streptococcus-treated macrophages.  

PubMed Central

Nitric oxide production by mouse macrophages treated with group B streptococci and gamma interferon was inhibited by cytochalasin B or by antibody neutralization of macrophage-derived tumor necrosis factor alpha. Phagocytosis-induced tumor necrosis factor alpha is responsible for group B streptococcus-induced nitric oxide production in interferon-treated macrophages. PMID:7642312

Goodrum, K J; Dierksheide, J; Yoder, B J

1995-01-01

338

Interleukin-1beta contributes via nitric oxide to the upregulation and functional activity of the zinc transporter Zip14 (Slc39a14) in murine hepatocytes.  

PubMed

Zinc metabolism during chronic disease is dysregulated by inflammatory cytokines. Experiments with IL-6 knockout mice show that LPS regulates expression of the zinc transporter, Zip14, by a mechanism that is partially independent of IL-6. The LPS-induced model of sepsis may occur by a mechanism signaled by nitric oxide (NO) as a secondary messenger. To address the hypothesis that NO can modulate Zip14 expression, we treated primary hepatocytes from wild-type mice with the NO donor S-nitroso N-acetyl penicillamine (SNAP). After treatment with SNAP, steady-state Zip14 mRNA levels displayed a maximal increase after 8 h and a concomitant increase in the transcriptional activity of the gene. Chromatin immunoprecipitation documented the kinetics of activator protein (AP)-1 and RNA polymerase II association with the Zip14 promoter after NO exposure, indicating a role of AP-1 in transcription of Zip14. We then stimulated the primary murine hepatocytes with IL-1beta, an LPS-induced proinflammatory cytokine and a potent activator of inducible NO synthase (iNOS) and NO production. In support of our hypothesis, IL-1beta treatment led to a threefold increase in Zip14 mRNA and enhanced zinc transport, as measured with a zinc fluorophore, in wild-type but not iNOS-/- hepatocytes. These data suggest that signaling pathways activated by NO are factors in the upregulation of Zip14, which in turn mediates hepatic zinc accumulation and hypozincemia during inflammation and sepsis. PMID:19179618

Lichten, Louis A; Liuzzi, Juan P; Cousins, Robert J

2009-04-01

339

Role of nitric oxide in activation of human T lymphocytes induced by bacterial superantigen.  

PubMed

The involvement of nitric oxide (NO) in the regulation of human T cell response to bacterial superantigen (staphylococcal enterotoxin B) was studied. It was shown that stimulated T lymphocytes are the main source of NO. This superantigen markedly increased NO production and triggered the proliferative response of mononuclear cells from healthy individuals; the degree of apoptosis was low. In patients with purulent surgical diseases with high spontaneous and induced NO production, superantigen enhanced apoptosis of lymphocytes and induced anergy of T cells to enterotoxins. Increasing the concentration of NO in cultured cells from healthy individuals in the presence of NO donors also stimulated apoptosis and inhibited proliferative activity. These data suggest that NO regulates T lymphocyte response to superantigens. The increased production of NO probably contributes to the development of immunosuppression during bacterial infection. PMID:11177292

Sakhno, L V; Leplina, O Y; Norkin, M N; Chernykh, E R; Ostanin, A A

2000-10-01

340

Inhibition of Lipopolysaccharide-Induced Cyclooxygenase2 and Inducible Nitric Oxide Synthase Expression by 4-[(2?-O-acetyl-?-L-Rhamnosyloxy)Benzyl]Isothiocyanate from Moringa oleifera  

Microsoft Academic Search

Moringa oleifera Lamarck is commonly consumed for nutritional or medicinal properties. We recently reported the isolation and structure elucidation of novel bioactive phenolic glycosides, including 4-[(2?-O-acetyl-?-L-rhamnosyloxy)benzyl]isothiocyanate (RBITC), which was found to suppress inducible nitric oxide synthase (iNOS) expression and nitric oxide (NO) production in lipopolysaccharide-stimulated RAW 264.7 mouse macrophage cells. Inhibitors of proteins such as cyclooxygenase-2 (COX-2) and iNOS are

Eun-Jung Park; Sarot Cheenpracha; Leng Chee Chang; Tamara P. Kondratyuk; John M. Pezzuto

2011-01-01

341

Dynamic compression counteracts IL1? induced inducible nitric oxide synthase and cyclo-oxygenase-2 expression in chondrocyte\\/agarose constructs  

Microsoft Academic Search

BACKGROUND: Nitric oxide and prostaglandin E2 (PGE2play pivotal roles in both the pathogenesis of osteoarthritis and catabolic processes in articular cartilage. These mediators are influenced by both IL-1? and mechanical loading, and involve alterations in the inducible nitric oxide synthase (iNOS) and cyclo-oxygenase (COX)-2 enzymes. To identify the specific interactions that are activated by both types of stimuli, we examined

TT Chowdhury; S Arghandawi; J Brand; OO Akanji; DL Bader; DM Salter; DA Lee

2008-01-01

342

Inhibition by parthenolide of phorbol ester-induced transcriptional activation of inducible nitric oxide synthase gene in a human monocyte cell line THP1  

Microsoft Academic Search

Excessive nitric oxide production by inducible nitric oxide synthase (iNOS) in stimulated inflammatory cells is thought to be a causative factor of cellular injury in inflammatory disease states. Compounds inhibiting iNOS transcriptional activity in inflammatory cells are potentially anti-inflammatory. An assay method for estimating iNOS transcriptional activity in the human monocyte cell line THP-1 was established using a luciferase reporter

Kazunori Fukuda; Yuko Hibiya; Michihiro Mutoh; Yasushi Ohno; Kazuya Yamashita; Seigou Akao; Hisayoshi Fujiwara

2000-01-01

343

Hypoxia-induced hypothermia mediated by noradrenaline and nitric oxide in the rostromedial preoptic area.  

PubMed

Central nitric oxide (NO) has an important role in hypothermia induced by hypoxia as well as in that elicited by noradrenaline (NA) microinjected into the rostromedial preoptic area (POA) of the hypothalamus. Here, I tested the hypothesis that activation of adrenoceptors and NO in the rostromedial POA is involved in hypoxia-induced hypothermia in urethane-chloralose-anesthetized, neuromuscularly blocked, artificially ventilated rats. Hypoxic ventilation (10% O2-90% N2, 5 min) evoked an increase in the tail skin temperature and a decrease in the colonic temperature, though these changes occurred at 30 s to 7 min after returning the rats to ventilation with room air. These responses were eliminated by prior bilateral transection of the carotid sinus nerves, but not by bilateral cervical vagotomy, suggesting the involvement of activated carotid chemoreceptors in the hypoxic ventilation-induced hypothermia. Such responses were also greatly attenuated by the microinjection of an NO synthase (NOS) inhibitor, NG-monomethyl-L-arginine (L-NMMA, 25 nmol), but not by that of its inactive enantiomer, NG-monomethyl-D-arginine (D-NMMA, 25 nmol), into the NA-sensitive, hypothermia-inducing site in the rostromedial POA. Pretreatment with the ?1-adrenoceptor blocker prazosin (50 pmol), but not vehicle saline, also greatly attenuated the hypoxic ventilation-induced heat loss responses. These results suggest that this hypoxia-induced hypothermia was mediated, at least in part, by activation of ?1-adrenoceptors and NOS in the rostromedial POA. PMID:21277355

Osaka, T

2011-04-14

344

Increased expression and cellular localization of inducible nitric oxide synthase and cyclooxygenase 2 in Helicobacter pylori gastritis  

Microsoft Academic Search

Background & Aims: Inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 are important regulators of mucosal inflammation and epithelial cell growth. To determine the role of iNOS and COX-2 in Helicobacter pylori–induced tissue injury, we compared their gene expression in H. pylori–induced gastritis with that in normal gastric mucosa and in non–H. pylori gastritis. Methods: In 43 patients, we assessed

Sidong Fu; Kalathur S. Ramanujam; Annie Wong; George T. Fantry; Cinthia B. Drachenberg; Stephen P. James; Stephen J. Meltzer; Keith T. Wilson

1999-01-01

345

Hypoxia augments lipopolysaccharide-induced cytokine expression in periodontal ligament cells.  

PubMed

Periodontitis is a chronic inflammatory disease characterized by the destruction of tooth supporting tissues. Hypoxia, the mainly changes of the plateau environment, can induce severe periodontitis by animal experiments. There is, however, very little information on hypoxia and lipopolysaccharide (LPS) induced cytokine expression in periodontal ligament (PDL) cells. In this article, we characterized hypoxia or P. gingivalis lipopolysaccharide (Pg LPS) induced tumor necrosis factor alpha (TNF-?), interleukin (IL)-1?, and IL-6 expression by human periodontal ligament (hPDL) cells. We found that hypoxia augmented Pg LPS induced TNF-?, IL-1?, and IL-6 expression in hPDL cells. We also demonstrated that nuclear factor kappa B pathway was involved in hypoxia augmenting Pg LPS induced cytokine expression in hPDL cells. Thus, our results suggest that the hypoxic environment may enhance the immune function of hPDL cells that is induced by Pg LPS. PMID:24609838

Jian, Congxiang; Li, Chenjun; Ren, Yu; He, Yong; Li, Yunming; Feng, Xiaodan; Zhang, Gang; Tan, Yinghui

2014-10-01

346

Nitric oxide contributes to substance P-induced increases in lung rapidly adapting receptor activity in guinea-pigs.  

PubMed Central

1. Substance P induces fluid flux via nitric oxide, and fluid flux stimulates lung rapidly adapting receptors (RARs). We therefore proposed that nitric oxide contributes to substance P-evoked increases in RAR activity. Since substance P decreases dynamic compliance (Cdyn), which can stimulate RARs, we also determined whether nitric oxide contributed to substance P-induced effects on pulmonary function. 2. In anaesthetized guinea-pigs, the effects of substance P on RAR activity, Cdyn, pulmonary resistance (RL), and arterial blood pressure were measured before and after i.v. infusion of NG-methyl-L-arginine (L-NMMA; a nitric oxide synthase inhibitor), or L-NMMA followed by L-arginine (a nitric oxide precursor which reverses the effects of L-NMMA). 3. Substance P-evoked increases in RAR activity were blunted by L-NMMA (P = 0.006) but not by L-NMMA-L-arginine (P = 0.42). 4. Substance P-evoked decreases in Cdyn were slightly inhibited by L-NMMA (P = 0.02) and slightly enhanced by L-NMMA-L-arginine (P = 0.004). However, at the time at which L-NMMA maximally reduced substance P-induced RAR stimulation (the first 30 s), it did not change substance P-induced decreases in Cdyn. 5. Substance P-evoked increases in RL were not changed by L-NMMA (P = 0.10) and were enhanced by L-NMMA-L-arginine (P = 0.03). 6. L-NMMA-evoked increases in mean arterial blood pressure were reversed by L-arginine. Substance P-evoked decreases in mean arterial blood pressure were not changed by L-NMMA or by L-NMMA-L-arginine. 7. We conclude that nitric oxide contributes to substance P-evoked increases in RAR activity and that the increases are most probably independent of decreases in Cdyn. PMID:9379417

Joad, J P; Kott, K S; Bonham, A C

1997-01-01

347

Differential cytokine expression in skin graft healing in inducible nitric oxide synthase knockout mice.  

PubMed

Inducible nitric oxide synthase (iNOS) and its product, nitric oxide, have been shown to play important roles in wound biology. The present study was performed to investigate the role of iNOS in modulating the cytokine cascade during the complex process of skin graft wound healing.Fifteen iNOS-knockout mice and 15 wild-type C57BL/6J mice were subjected to autogenous 1-cm2 intrascapular full-thickness skin grafts. Three animals in each group were killed on postoperative days 3, 5, 7, 10, and 14. Specimens were then analyzed using nonisotopic in situ hybridization versus mRNA of tumor growth factor-beta1, vascular endothelial growth factor, iNOS, endothelial nitric oxide synthase (eNOS), tumor necrosis factor-alpha, and basic fibroblast growth factor, as well as positive and negative control probes. Positive cells in both grafts and wound beds were counted using a Leica microgrid. Scar thickness was measured with a Leica micrometer. Data were analyzed using the unpaired Student's t test. Expression of iNOS was 2- to 4-fold higher in knockout mice than in wild-type mice on postoperative days 5, 7, and 14. Expression of eNOS was 2- to 2.5-fold higher in knockout mice than in wild-type mice on postoperative days 5 and 7. Tumor necrosis factor-alpha expression was 2- to 7-fold higher in knockout mice than in wild-type mice on all postoperative days. In contrast, expression levels of angiogenic/fibrogenic cytokines (vascular endothelial growth factor, basis fibroblast growth factor, and tumor growth factor-beta1) were 2.5- to 4-fold higher in wild-type mice than in knockout mice. Scars were 1.5- to 2.5-fold thicker in knockout mice than in wild-type mice at all time points. All of the above results represent statistically significant differences (p < 0.05). Significantly different patterns of cytokine expression were seen in knockout and wild-type mice. Although the scar layer was thicker in knockout mice, it showed much greater infiltration with inflammatory cells. These data further delineate the modulatory effect of iNOS and nitric oxide in healing skin grafts. PMID:11604628

Most, D; Efron, D T; Shi, H P; Tantry, U S; Barbul, A

2001-10-01

348

Nitric oxide protects anterior pituitary cells from cadmium-induced apoptosis.  

PubMed

Cadmium (Cd2+) is a potent toxic metal for both plants and animals. Chronic exposure to low doses of Cd2+ results in damage to several organs. We have previously reported that Cd2+ induces apoptosis in anterior pituitary cells by a caspase- and oxidative stress-dependent mechanism. Nitric oxide (NO) synthesis is affected by Cd2+ in several systems. NO has been shown to be either cytoprotective or cytotoxic in many systems. The aim of this study was to evaluate the possible participation of NO in the cytotoxic effect of Cd2+ on rat anterior pituitary cells. Cell viability was evaluated by mitochondrial dehydrogenase activity assay and confirmed by microscopy, studying nuclear morphology. Here we show that DETA NONOate ((Z)-1-[2 (2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate), a long-term NO donor, at concentrations below 0.5 mM, reduces nuclear condensation and fragmentation and reverses the decrease in cellular activity induced by Cd2+. Cd2+, by itself, induced NO synthesis, and inhibition of this synthesis enhanced Cd2+ cytotoxicity. NO also prevented caspase-3 activation and lipidic peroxidation induced by Cd2+. The NO/cGMP pathway does not seem to be involved in the cytoprotective effect of NO. These results indicate that NO has a cytoprotective role in Cd2+ -induced apoptosis, suggesting that endogenous NO could have a physiological role in protecting anterior pituitary cells. PMID:15454286

Poliandri, Ariel H B; Velardez, Miguel O; Cabilla, Jimena P; Bodo, Cristian C A; Machiavelli, Leticia I; Quinteros, Alnilan F; Duvilanski, Beatriz H

2004-11-01

349

Modulation of picryl chloride-induced contact hypersensitivity reaction in mice by nitric oxide.  

PubMed

We have studied the possible involvement of nitric oxide (NO) in the contact hypersensitivity reaction. A biphasic response of ear swelling was observed at 2 h (early phase) and 24 h (late phase) after application of the antigen to picryl chloride (PC1)-sensitized CBA/J mice. Intravenous injection of NO synthase inhibitor, NG-nitro-L-arginine methyl ester (L-NAME), at the time of PC1 challenge, inhibited in a concentration-dependent fashion the antigen-induced contact hypersensitivity reaction. Low-dose (1 mg/kg) L-NAME inhibited the early-phase reaction but not the late-phase reaction. High-dose (250 mg/kg) L-NAME inhibited both early- and late-phase reactions. D-NAME (enantiomer of L-NAME) did not inhibit the antigen-induced ear swelling. High-dose (250 mg/kg) L-arginine increased both early and late phase reactions. D-Arginine (enantiomer of L-arginine) did no increase the antigen-induced ear swelling. L-NAME injection, however, did not suppress phenol-induce irritant inflammation. Treatment of mice undergoing PC1-induced contact hypersensitivity reaction with L-NAME reduced the production of interleukin-2 and interferon-gamma by draining lymph node cells. Treatment with L-arginine, on the other hand increased the production of interleukin-2 and interferon-gamma. These results suggest that NO plays a modulating role in contact hypersensitivity reaction. PMID:8823359

Morita, H; Hori, M; Kitano, Y

1996-10-01

350

Influence of lysophosphatidic acid on nitric oxide-induced luteolysis in steroidogenic luteal cells in cows.  

PubMed

Lysophosphatidic acid (LPA) together with its active G protein-coupled receptors are present in the corpus luteum (CL) of the cow. Under in vivo conditions, LPA stimulated P4 and PGE2 secretion during the luteal phase of the estrous cycle in heifers. Furthermore, LPA maintained P4 synthesis and actions in the bovine CL in vitro. However, the effect of this phospholipid on nitric oxide (NO)-induced functional and structural luteolysis has not been investigated. The aim of the present work was to determine the effects of LPA on 1) NO-induced functional luteolysis, 2) NO-dependent PG synthesis, and 3) NO-induced structural luteolysis in cultured steroidogenic luteal cells. We documented that LPA reversed the inhibitory effect of NONOate, an NO donor, on P4 synthesis and PGE2/PGF2alpha ratio in cultured steroidogenic luteal cells. Additionally, LPA inhibited NO-induced apoptosis in cultured steroidogenic luteal cells via abrogation of the NO-dependent stimulatory influence on proapoptotic TNFalpha/TNFR1 and Fas/FasL expression, Caspase 3 activity, and the Bax/Bcl2 ratio during luteal regression in the bovine CL. In conclusion, this study proves that in the presence of LPA, NO cannot induce luteolytic capacity acquisition, leading to functional and structural luteolysis of bovine luteal cells. PMID:24307705

Kowalczyk-Zieba, Ilona; Boruszewska, Dorota; Sinderewicz, Emilia; Skarzynski, Dariusz Jan; Woclawek-Potocka, Izabela

2014-01-01

351

The effects of nicotine on nitric oxide induced anxiogenic-like behaviors in the dorsal hippocampus.  

PubMed

Nicotine, an active tobacco derived alkaloid, regulates the activity of the neuronal nitric oxide synthase (nNOS) as well as the release of nitric oxide (NO) in the nervous system. Nicotinic acetylcholine receptors and nNOS are abundantly co-expressed in the hippocampal neurons and are found to alter anxiety-like behaviors in rodents. Dorsal hippocampus may be a site for modulation of anxiety. Therefore, in this study, we investigated the possible interactions between nicotine and NO systems of the dorsal hippocampus and the resultant effect on anxiety-like behaviors. The elevated plus-maze (EPM) test has been used to test the anxiety. Intraperitoneal administration of nicotine (0.5mg/kg) decreased the open arm time percentage (%OAT) and open arm entries percentage (%OAE) but not the locomotor activity, indicating an anxiogenic-like response. Intra-CA1 injection of l-arginine (a NO precursor) or l-NAME (a NOS inhibitor) also caused anxiogenic-like effects. On the other hand, injection of the low dose nicotine before different doses of l-arginine or l-NAME blocked the anxiogenic-like response induced by the drugs. Our results suggested that, both NO and nicotinic cholinergic systems not only play a part in the modulation of the anxiety in mice dorsal hippocampus, but also demonstrate a complex interaction in this respect. PMID:22985517

Piri, Morteza; Nasehi, Mohammad; Shahab, Zahra; Zarrindast, Mohammad Reza

2012-10-24

352

Renal angiotensin-converting enzyme is essential for the hypertension induced by nitric oxide synthesis inhibition.  

PubMed

The kidney is an important source of angiotensin-converting enzyme (ACE) in many species, including humans. However, the specific effects of local ACE on renal function and, by extension, BP control are not completely understood. We previously showed that mice lacking renal ACE, are resistant to the hypertension induced by angiotensin II infusion. Here, we examined the responses of these mice to the low-systemic angiotensin II hypertensive model of nitric oxide synthesis inhibition with L-NAME. In contrast to wild-type mice, mice without renal ACE did not develop hypertension, had lower renal angiotensin II levels, and enhanced natriuresis in response to L-NAME. During L-NAME treatment, the absence of renal ACE was associated with blunted GFR responses; greater reductions in abundance of proximal tubule Na(+)/H(+) exchanger 3, Na(+)/Pi co-transporter 2, phosphorylated Na(+)/K(+)/Cl(-) cotransporter, and phosphorylated Na(+)/Cl(-) cotransporter; and greater reductions in abundance and processing of the ? isoform of the epithelial Na(+) channel. In summary, the presence of ACE in renal tissue facilitates angiotensin II accumulation, GFR reductions, and changes in the expression levels and post-translational modification of sodium transporters that are obligatory for sodium retention and hypertension in response to nitric oxide synthesis inhibition. PMID:25012170

Giani, Jorge F; Janjulia, Tea; Kamat, Nikhil; Seth, Dale M; Blackwell, Wendell-Lamar B; Shah, Kandarp H; Shen, Xiao Z; Fuchs, Sebastien; Delpire, Eric; Toblli, Jorge E; Bernstein, Kenneth E; McDonough, Alicia A; Gonzalez-Villalobos, Romer A

2014-12-01

353

Expression of inducible nitric oxide synthase and its involvement in pulmonary granulomatous inflammation in rats.  

PubMed Central

Two types of pulmonary granulomatosis were produced in rats by intratracheal instillation of zymosan or silica. In both models, immunostaining with anti-rat monoclonal antibody for inducible nitric oxide synthase (iNOS), ANOS11, showed that the intensity of iNOS immunoreactivity in the inflammatory lesions peaked at 3 days and declined thereafter. Immunohistochemical double staining and in situ hybridization demonstrated the expression of iNOS in neutrophils, monocyte-derived macrophages, and bronchiolar epithelial cells in the pulmonary lesions. Electron spin resonance spectroscopy revealed the production of an excessive amount of nitric oxide (NO) in the pulmonary lesions. Immunostaining with a polyclonal antibody against nitrotyrosine indicated the formation of nitrotyrosine residues in the granulomatous lesions, particularly in the periphery of the lesions, providing indirect evidence for the generation of peroxynitrite anion in the zymosan- or silica-instilled lungs. Administration of N omega-nitro-L-arginine methyl ester or S-methylisothiourea sulfate, which significantly suppressed NO production, resulted in marked reduction of monocyte/macrophage infiltration as well as in inhibition of induction of monocyte chemoattractant protein-1 in the lesions. These data indicate that NO and its more reactive product peroxynitrite anion may be important mediators of granuloma formation in the lung. Images Figure 1 Figure 2 Figure 4 Figure 8 Figure 9 PMID:8952535

Setoguchi, K.; Takeya, M.; Akaike, T.; Suga, M.; Hattori, R.; Maeda, H.; Ando, M.; Takahashi, K.

1996-01-01

354

Inducible and endothelial nitric oxide synthase expression during development of transplant arteriosclerosis in rat aortic grafts.  

PubMed Central

In the vascular system, distinct isoforms of nitric oxide synthase (NOS) generate nitric oxide (NO), which acts as a biological messenger. Its role in the development of transplant arteriosclerosis (TA) is still unclear. To investigate whether NO is involved in TA, we studied the expression of NOS isoforms, inducible NOS (iNOS) and endothelial NOS (eNOS), by immunohistochemistry and in situ hybridization during the first two post-transplantation months and their relation with cold ischemia (1 to 24 hours) and reperfusion injury using an aortic transplantation model in the rat. We found an increased iNOS expression in the intima and adventitia and a decreased expression in the media, whereas eNOS expression was not significantly altered during the development of TA. Co-localization studies suggested that iNOS-positive cells were vascular smooth muscle cells, monocyte-derived macrophages, and endothelial cells. Prolonged ischemic storage time resulted in an increase in eNOS expression in the neointima. In situ hybridization showed iNOS mRNA expression by vascular cells in the neointima and media. NO produced by iNOS and eNOS may be involved, at least in part, in the pathogenesis of TA in aortic grafts. Additional studies are needed to confirm the modulatory mechanism of NO during the development of TA. Images Figure 3 Figure 4 Figure 6 PMID:8952533

Akyürek, L. M.; Fellström, B. C.; Yan, Z. Q.; Hansson, G. K.; Funa, K.; Larsson, E.

1996-01-01

355

Chitosan and blueberry treatment induces arginase activity and inhibits nitric oxide production during acetaminophen-induced hepatotoxicity  

PubMed Central

Background: Liver diseases have become a major problem of the worldwide. More than 50% of all cases of liver failure can be attributed to drugs. Among these, acetaminophen is the most common cause. Objective: The aim of this study was to investigate the the hepatoprotective effects of blueberry and chitosan on tissue arginase activity, ornithine and nitric oxide levels during the acetaminophen-induced hepatotoxicity. Materials and Methods: Acetaminophen (250 mg/kg body weight per day), blueberry (60 mg/kg body weight per day) and, chitosan (200 mg/kg body weight per day) were administered to the rats by oral gavage during the experimental period. Results: Blueberry and chitosan significantly decreased liver arginase activity and ornithine levelsand and increased nitric oxide levels. Glutathione levels were remarkably increased by chitosan and blueberry treatments. Conclusion: The results of the present study indicate that blueberry and chitosan effectively protected against the acetaminophen-induced hepatotoxicity. The hepatoprotective effect afforded by blueberry and chitosan can be attributed to its antioxidant and anti-inflammatory activities. PMID:24991095

Ozcelik, Eda; Uslu, Sema; Burukoglu, Dilek; Musmul, Ahmet

2014-01-01

356

Effect of 17?-oestradiol on cytokine-induced nitric oxide production in rat isolated aorta  

PubMed Central

Studies were performed on isolated aortic rings without endothelium to investigate the effect of 17?-oestradiol on cytokine-induced nitric oxide production by the inducible nitric oxide synthase (iNOS).Treatment of the isolated aortic rings with interleukin-1? (IL-1?, 20???ml?1) led to the expression of iNOS mRNA and protein, as well as significant nitrite accumulation in the incubation media and suppression of phenylephrine (1?nM–10??M)-evoked contraction.Cycloheximide (1??M), a protein synthesis inhibitor, prevented iNOS protein expression, nitrite accumulation and the suppression of contractility by IL-1? on the isolated aortic rings. 17?-oestradiol (1?nM–10??M) and the partial oestrogen receptor agonist 4-OH-tamoxifen (1?nM–10??M) produced concentration-dependent inhibition of IL-1?-induced nitrite accumulation and restored vasoconstrictor responsiveness to phenylephrine, similar to the iNOS inhibitor aminoguanidine (100??M).Semiquantitative PCR demonstrated decreased iNOS mRNA in the IL-1?-induced and 17?-oestradiol-treated rings. Western blot analysis of rat aorta homogenates revealed that 17?-oestradiol treatment resulted in a reduction in IL-1ß-induced iNOS protein level.Incubation with tumour necrosis factor ? (TNF?, 1?ng?ml?1) resulted in significant nitrite accumulation in the incubation media and suppression of the smooth muscle contractile response to phenylephrine, similar to IL-1?. The effects of TNF? were also inhibited by co-incubation of the rings with 17?-oestradiol and 4-OH-tamoxifen (1??M).The anti-transforming growth factor-?1 (TGF-?1) antibody, which inhibited TGF-?1-induced suppression of nitrite production from IL-1?-treated vascular rings, did not affect the inhibitory action of 17?-oestradiol, suggesting that the effect of oestrogen on iNOS inhibition was not mediated by TGF-?1.These results show that the ovarian sex steroid, 17?-oestradiol is a modulator of cytokine-induced iNOS activity in rat vascular smooth muscle and its mechanism of action involves decrease of iNOS mRNA and protein. PMID:9559891

Kauser, Katalin; Sonnenberg, Dagmar; Diel, Patrick; Rubanyi, Gabor M

1998-01-01

357

Nitric Oxide Stress Induces Different Responses but Mediates Comparable Protein Thiol Protection in Bacillus subtilis and Staphylococcus aureus  

Microsoft Academic Search

The nonpathogenic Bacillus subtilis and the pathogen Staphylococcus aureus are gram-positive model organ- isms that have to cope with the radical nitric oxide (NO) generated by nitrite reductases of denitrifying bacteria and by the inducible NO synthases of immune cells of the host, respectively. The response of both microor- ganisms to NO was analyzed by using a two-dimensional gel approach.

Falko Hochgrafe; Carmen Wolf; Stephan Fuchs; Manuel Liebeke; Michael Lalk; Susanne Engelmann; Michael Hecker

2008-01-01

358

Synergy between ethanol and grape polyphenols, quercetin, and resveratrol, in the inhibition of the inducible nitric oxide synthase pathway  

Microsoft Academic Search

In atherosclerosis and tumor initiation, inducible nitric oxide synthase (iNOS) has been implicated in the damage of vascular walls and DNA, respectively. Moderate consumption of red wine has been ascribed as a preventive for coronary heart disease; however, there has been much debate over whether the beneficial effect is from grape polyphenolic components or ethanol. We studied the interaction of

Marion Man-Ying Chan; John A Mattiacci; Hyon S Hwang; Amit Shah; Dunne Fong

2000-01-01

359

Implication of NK1 and NK2 receptors in rat colonic hypersecretion induced by interleukin 1?: Role of nitric oxide  

Microsoft Academic Search

Background & Aims: Interleukin (IL) 1? is known to induce a neurally mediated colonic water secretion in vivo. The aim of this study was to investigate the mechanism of action of IL-1? on colonic net water flux and the role of tachykinins and nitric oxide. Methods: In anesthetized rats, isolated colonic loops were infused with Ringer's buffer containing [14C]polyethylene glycol

Helene Eutamene; Vassilia Theodorou; Jean Fioramonti; Lionel Bueno

1995-01-01

360

Poster Session 08: Bystander and other Low Dose Effect Roles of nitric oxide in adaptive response induced in zebrafish  

E-print Network

induced in zebrafish embryos in vivo by microbeam protons Viann Wing Yan CHOI1, Candy Yuen Ping NG1, Alisa (5 h post-fertilization, hpf) embryos of the zebrafish, Danio rerio, by 3.4 MeV protons from; adaptive response; nitric oxide; zebrafish embryos REFERENCE 1. Konishi T, Oikawa M, Suya N et al. SPICE

Yu, K.N.

361

Inducible nitric oxide synthase (iNOS) expression may predict distant metastasis in human melanoma  

PubMed Central

Expression of inducible nitric oxide synthase (iNOS) and its cellular localization was investigated in subcutaneous or lymph node metastases of human melanoma. Immunohistochemistry revealed that iNOS expression was limited to melanoma cells. In samples of patients without distant metastases, the number of iNOS+ tumour cells/total tumour cells was 55% ± 17% (n = 12) compared with 9% ± 8% when distant metastases of lung, liver or brain occurred within an observation period of 3 years (n = 10) (P < 0.001). Western blotting confirmed the expression of iNOS protein in select cases. Notably, iNOS is expressed in regional melanoma metastases and its expression is inversely related to the tumour's metastatic potential. Thus, iNOS expression may have predictive value for the development of distant metastases of human melanoma. © 1999 Cancer Research Campaign PMID:10188914

Tschugguel, W; Pustelnik, T; Lass, H; Mildner, M; Weninger, W; Schneeberger, C; Jansen, B; Tschachler, E; Waldhör, T; Huber, J C; Pehamberger, H

1999-01-01

362

Citrus nobiletin suppresses inducible nitric oxide synthase gene expression in interleukin-1?-treated hepatocytes  

SciTech Connect

Highlights: •Nobiletin is a polymethoxylated flavone that is abundant in citrus peels. •Nobiletin is a major constituent of the Citrus unshiu peel extract. •Nobiletin suppresses induction of NO and reduces iNOS expression in hepatocytes. •Nobiletin reduces the iNOS promoter activity and the DNA-binding activity of NF-?B. -- Abstract: Background: Nobiletin is a polymethoxylated flavone that is abundant in the peels of citrus fruits, such as Citrus unshiu (Satsuma mandarin) and Citrus sinensis. The dried peels of C. unshiu (chinpi) have been included in several formulae of Japanese Kampo medicines. Nobiletin may suppress the induction of inducible nitric oxide synthase (iNOS), which synthesizes the inflammatory mediator nitric oxide (NO) in hepatocytes. Methods: A C. unshiu peel (CUP) extract was prepared. Primary cultured rat hepatocytes were treated with the CUP extract or nobiletin in the presence of interleukin 1? (IL-1?), which induces iNOS expression. NO production and iNOS gene expression were analyzed. Results: High-performance liquid chromatography analyses revealed that the nobiletin content in the CUP extract was 0.14%. Nobiletin dose-dependently reduced the NO levels and decreased iNOS expression at the protein, mRNA and antisense transcript levels. Flavone, which does not contain any methoxy groups, also suppressed iNOS induction. Nobiletin reduced the transcriptional activity of iNOS promoter-luciferase constructs and the DNA-binding activity of nuclear factor ?B (NF-?B) in the nuclei. Conclusions: The suppression of iNOS induction by nobiletin suggests that nobiletin may be responsible for the anti-inflammatory effects of citrus peels and have a therapeutic potential for liver diseases.

Yoshigai, Emi [Department of Biomedical Sciences, College of Life Sciences, Kusatsu, Shiga (Japan) [Department of Biomedical Sciences, College of Life Sciences, Kusatsu, Shiga (Japan); Ritsumeikan Global Innovation Research Organization (R-GIRO), Kusatsu, Shiga (Japan); Machida, Toru [Department of Biomedical Sciences, College of Life Sciences, Kusatsu, Shiga (Japan)] [Department of Biomedical Sciences, College of Life Sciences, Kusatsu, Shiga (Japan); Okuyama, Tetsuya [Ritsumeikan Global Innovation Research Organization (R-GIRO), Kusatsu, Shiga (Japan)] [Ritsumeikan Global Innovation Research Organization (R-GIRO), Kusatsu, Shiga (Japan); Mori, Masatoshi; Murase, Hiromitsu; Yamanishi, Ryota [Department of Biomedical Sciences, College of Life Sciences, Kusatsu, Shiga (Japan)] [Department of Biomedical Sciences, College of Life Sciences, Kusatsu, Shiga (Japan); Okumura, Tadayoshi [Research Organization of Science and Technology, Ritsumeikan University, Kusatsu, Shiga (Japan) [Research Organization of Science and Technology, Ritsumeikan University, Kusatsu, Shiga (Japan); Department of Surgery, Kansai Medical University, Hirakata, Osaka (Japan); Ikeya, Yukinobu [Department of Pharmacy, College of Pharmaceutical Sciences, Ritsumeikan University, Kusatsu, Shiga (Japan)] [Department of Pharmacy, College of Pharmaceutical Sciences, Ritsumeikan University, Kusatsu, Shiga (Japan); Nishino, Hoyoku [Ritsumeikan Global Innovation Research Organization (R-GIRO), Kusatsu, Shiga (Japan) [Ritsumeikan Global Innovation Research Organization (R-GIRO), Kusatsu, Shiga (Japan); Department of Biochemistry, Kyoto Prefectural University of Medicine, Kyoto (Japan); Nishizawa, Mikio, E-mail: nishizaw@sk.ritsumei.ac.jp [Department of Biomedical Sciences, College of Life Sciences, Kusatsu, Shiga (Japan)] [Department of Biomedical Sciences, College of Life Sciences, Kusatsu, Shiga (Japan)

2013-09-13

363

Nitric oxide and hypoxia exacerbate alcohol-induced mitochondrial dysfunction in hepatocytes  

PubMed Central

Chronic alcohol consumption results in hepatotoxicity, steatosis, hypoxia, increased expression of inducible nitric oxide synthase (iNOS) and decreased activities of mitochondrial respiratory enzymes. The impact of these changes on cellular respiration and their interaction in a cellular setting is not well understood. In the present study we tested the hypothesis that nitric oxide (?NO)-dependent modulation of cellular respiration and the sensitivity to hypoxic stress is increased following chronic alcohol consumption. This is important since ?NO has been shown to regulate mitochondrial function through its interaction with cytochrome c oxidase, although at higher concentrations, and in combination with reactive oxygen species, can result in mitochondrial dysfunction. We found that hepatocytes isolated from alcohol-fed rats had decreased mitochondrial bioenergetic reserve capacity and were more sensitive to ?NO-dependent inhibition of respiration under room air and hypoxic conditions. We reasoned that this would result in greater hypoxic stress in vivo, and to test this, wild-type and iNOS?/? mice were administered alcohol-containing diets. Chronic alcohol consumption resulted in liver hypoxia in the wild-type mice and increased levels of hypoxia-inducible factor 1? in the peri-venular region of the liver lobule. These effects were attenuated in the alcohol-fed iNOS?/? mice suggesting that increased mitochondrial sensitivity to ?NO and reactive nitrogen species in hepatocytes and iNOS plays a critical role in determining the response to hypoxic stress in vivo. These data support the concept that the combined effects of ?NO and ethanol contribute to an increased susceptibility to hypoxia and the deleterious effects of alcohol consumption on liver. PMID:21971515

Zelickson, Blake R.; Benavides, Gloria A.; Johnson, Michelle S.; Chacko, Balu K.; Venkatraman, Aparna; Landar, Aimee; Betancourt, Angela M.; Bailey, Shannon M.; Darley-Usmar, Victor M.

2011-01-01

364

A TLR4\\/MD2 fusion protein inhibits LPS-induced pro-inflammatory signaling in hepatic stellate cells  

Microsoft Academic Search

Activated hepatic stellate cells (HSCs) play a key role in hepatic fibrogenesis. In injured liver they are the main extracellular matrix protein producing cell type and further perpetuate hepatic injury by secretion of pro-inflammatory mediators. Since LPS-mediated signaling through toll-like receptor 4 (TLR4) has been identified as key fibrogenic signal in HSCs we aimed to test TLR4 as potential target

Bernd Schnabl; Katharina Brandl; Marina Fink; Philipp Gross; Kojiro Taura; Erwin Gaebele; Claus Hellerbrand; Werner Falk

2008-01-01

365

Acrolein with an alpha, beta-unsaturated Carbonyl Group Inhibits LPS-induced Homodimerization of Toll-like Receptor 4  

Technology Transfer Automated Retrieval System (TEKTRAN)

Acrolein is a highly electrophilic a,ß-unsaturated aldehyde present in a number of environmental sources, especially cigarette smoke. It reacts strongly with the thiol groups of cysteine residues by Michael addition and has been reported to inhibit nuclear factor-kB (NF-kB) activation by lipopolysac...

366

Abies koreana Essential Oil Inhibits Drug-Resistant Skin Pathogen Growth and LPS-Induced Inflammatory Effects of Murine Macrophage  

Microsoft Academic Search

Since acne vulgaris is the combined result of a bacterial infection and the inflammatory response to that infection, we examined\\u000a whether Abies koreana essential oil (AKE) possessed anti-inflammatory and antibacterial activities against skin pathogens. In this study, AKE showed\\u000a excellent antibacterial activities against drug-susceptible and -resistant Propionibacterium acnes and Staphylococcus epidermidis, which are acne-causing bacteria. In addition, AKE reduced the

Weon-Jong Yoon; Sang-Suk Kim; Tae-Heon Oh; Nam Ho Lee; Chang-Gu Hyun

2009-01-01

367

Knockdown of interleukin-1? does not attenuate LPS-induced production of interleukin-1? in mouse macrophages.  

PubMed

IL-1? and IL-1? are synthesized as 31kDa cell-associated precursors following TLR-4 stimulation, but their processing to the mature form and secretion require a second intracellular stimulus. The unique localization of the precursor of IL-1? (pro-IL-1?) to the nucleus suggested a role in transcriptional regulation of inflammatory cytokines. We explored the hypothesis that pro-IL-1? is involved in regulation of IL-1? expression following TLR-4 stimulation. IL-1? mRNA and protein levels were specifically decreased in macrophages from IL-1?-deficient mice following TLR-1/2, TLR-4 or TLR-9 stimulation, supporting the hypothesis. However, activation of the main upstream regulators of IL-1? expression, IRF3, NFkB and p38/JNK, were not reduced in macrophages from IL-1?-deficient mice. In order to assess the specific role of IL-1? in macrophages, we generated mice with myeloid cell deficiency of IL-1? (LyzMCre-loxp). Despite over 90% knockdown of IL-1?, TLR-4 stimulated macrophages from LyzMCre-loxp mice did not produce lower levels of IL-1? compared to IL-1?-loxp-flanked mice. In order to overcome the possibility that effects are caused by the incomplete deficiency of IL-1?, we generated new whole-body IL-1? knockout mice (GeneralCre-IL-1?) and the findings were similar to myeloid cell-deficient IL-1?. Collectively, our findings do not support the previously suggested role of nuclear IL-1? in gene regulation of IL-1?. Rather, they suggest that IL-1? acts mainly as an alarmin that is sequestered in the nucleus following stimulation with TLR-4. PMID:25748836

Almog, Tal; Kandel-Kfir, Michal; Shaish, Aviv; Dissen, Moshe; Shlomai, Gadi; Voronov, Elena; Apte, Ron N; Harats, Dror; Kamari, Yehuda

2015-05-01

368

Inhibition of inflammatory response in LPS-induced macrophages by 9-KOTE and 13-KOTE produced by biotransformation.  

PubMed

Lipid mediators such as the leukotrienes, resolvins and protectins have been considered excellent models for the development of new anti-inflammatory drugs, due to their high potentiality. Nevertheless, only tiny amounts are available from natural sources and they have to be prepared by total synthesis. It is known that besides chemical reagents, microorganisms can also promote fatty acid oxygenation, via enzymatic reactions. In this context, the aim of this work was to produce oxylipids analogues in structure to lipid mediators employing microbial biotransformation. To this end, ?-linolenic acid (ALA) was biotransformed by the fungi Aspergillus niger into oxylipids with different levels of oxygenation within 24h or 48h. The anti-inflammatory potential of products were evaluated by means of NO and TNF-? quantification in LPS-stimulated RAW264.7 macrophage cell line which guided the isolation of the regioisomers at m/z [M-H](-) 291, 9-keto-10E,12Z,15Z-octadecatrienoic acid (9-KOTE) and 13-keto-9Z,11E,15Z-octadecatrienoic acid (13-KOTE). We showed that biotransformation represents a powerful strategy for the production of potentially interesting candidates for development of anti-inflammation therapies. PMID:24731823

Petta, Tânia; Secatto, Adriana; Faccioli, Lúcia Helena; Moraes, Luiz Alberto Beraldo

2014-05-10

369

Parthenolide attenuates LPS-induced activation of NF-?B in a time-dependent manner in rat myocardium?  

PubMed Central

Parthenolide (PTN), a selective nuclear factor kappa B (NF-?B) inhibitor, has been used extensively to inhibit NF-?B activation. The duration of the inhibitory effect of PTN on NF-?B in vivo remains unclear. This study was to determine whether a lipopolysaccharide (LPS) challenge 6, 12 and 24 h after the administration of PTN could activate NF-?B. Rats were devided into five groups. The rats in the PTN, PTN+LPS and DMSO groups were injected intraperitoneally with PTN or DMSO. After 6, 12 or 24 h, LPS was administered in LPS and PTN+LPS groups. The expressions of NF-?B p50, I?B? and p-I?B? were inhibited in both PTN and PTN+LPS group at end of 6 and 12 h and no effects at 24 h. In summary, myocardial NF-?B expression occurs 1 h after the administration of LPS. PTN blocks this effect given at 6 h and no inhibitory effect 24 h after administration in vivo. PMID:23554728

Xie, Hong; Wang, Chen; Wu, Xuemei; Liu, Xia; Qiao, Shigang; Liu, Chunfeng; Liu, Hong

2012-01-01

370

A low-level diode laser therapy reduces the lipopolysaccharide (LPS)-induced periodontal ligament cell inflammation  

NASA Astrophysics Data System (ADS)

The purpose of this study was to investigate the cytologic effects of inflammatory periodontal ligament cells in vitro after low-level laser therapy. Human periodontal ligament cells were cultured, exposed to lipopolysaccharide and subjected to low-level laser treatment of 5?J?cm?2 or 10?J?cm?2 using a 920?nm diode laser. A periodontal ligament cell attachment was observed under a microscope, and the cell viability was quantified by a mitochondrial colorimetric assay. Lipopolysaccharide-treated periodontal ligament cells were irradiated with the low-level laser, and the expression levels of several inflammatory markers, iNOS, TNF-? and IL-1, and pErk kinase, were analyzed by reverse transcription polymerase chain reaction and western blot. The data were collected and analyzed by one-way analysis of variance; p < 0.05 indicated a statistically significant difference. The low-level laser treatment of periodontal ligament cells increased their ability to attach and survive. After irradiation, the expression levels of iNOS, TNF-? and IL-1 in lipopolysaccharide-exposed periodontal ligament cells decreased over time (p < 0.05). In periodontal ligament cells, low-level diode laser treatment increased the cells’ proliferative ability and decreased the expression of the examined inflammatory mediators.

Huang, T. H.; Chen, C. C.; Liu, S. L.; Lu, Y. C.; Kao, C. T.

2014-07-01

371

Impaired LPS-induced signaling in microglia overexpressing the Wiskott Aldrich syndrome protein N-terminal domain  

Microsoft Academic Search

Wiskott-Aldrich syndrome protein (WASP) plays important roles in TCR signaling, but its roles in signal transduction in innate immune cells have not been well characterized. As microglia are the primary immune effector cells in the brain, WASP may possibly have important roles in microglial activation, such as production of inflammatory and anti-inflammatory cytokines and neurotoxic factors. Here, we established a

Mitsuru Sato; Kazumasa Ogihara; Ryoko Sawahata; Kenji Sekikawa; Hiroshi Kitani

2007-01-01

372

Activation of the bile acid receptor TGR5 enhances LPS-induced inflammatory responses in a human monocytic cell line.  

PubMed

Abstract Introduction: Bile acids are recognized as signaling molecules, mediating their effects both through the cell surface receptor TGR5 and the nuclear receptor FXR. After a meal, approximately 95% of the bile acids are transported from terminal ileum and back to the liver via the portal vein, resulting in postprandial elevations of bile acids in blood. During the digestion of fat, components from the microbiota, including LPS, are thought to reach the circulation where it may lead to inflammatory responses after binding TLR4 immune cells. Both LPS and bile acids are present in blood after a high-fat meal; we therefore wanted to study consequences of a possible interplay between TGR5 and TLR4 in human monocytes. Methods: The monocytic cell line U937 stably transfected with the NF-?B reporter plasmid 3x-?B-luc was used as a model system to study the effects of TGR5 and TLR4. Activation of MAP kinases was studied to reveal functional consequences of triggering TGR5 in U937 cells. Effects of TGR5 and TLR4 activation were monitored using NF-?B luciferase assay and by quantification of the pro-inflammatory cytokines IL-6 and IL-8 using ELISA. Results: In this study, results show that triggering TGR5 with the specific agonist betulinic acid (BA), and the bile acids CDCA or DCA, activated both the main MAP kinases ERK1/2, p38 and JNK, and the NF-?B signaling pathway. We further demonstrated that co-triggering of TLR4 and TGR5 enhanced the activation of NF-?B and the release of inflammatory cytokines in a synergistic manner compared to triggering of TLR4 alone. Conclusions: Thus, two different and simultaneous events associated with the digestive process coordinately affect the function of human monocytes and contribute to enhanced inflammation. Because elevated levels of circulatory LPS may contribute to the development of insulin resistance, the results from this study suggest that bile acids through the activation of TGR5 may have a role in the development of insulin resistance as well. PMID:25418122

Mobraten, Kaia; Haugbro, Tarjei; Karlstrom, Ellen; Kleiveland, Charlotte R; Lea, Tor

2014-11-24

373

Chrysoeriol potently inhibits the induction of nitric oxide synthase by blocking AP-1 activation.  

PubMed

Chrysoeriol is a flavonoid with antioxidant and anti-inflammatory activities. Despite the large number of studies performed on its biological activities, no clear picture of its mode of action has emerged. In the present study, we isolated chrysoeriol from the leaves of Digitalis purpurea (foxglove), and studied its effect on the induction of the inducible nitric oxide synthase (iNOS) gene, and the mechanism of this induction in Raw264.7 macrophages. Chrysoeriol pretreatment potently inhibited the release of NO in the cells treated with lipopolysaccharide (LPS), and Western blot and RT-PCR analyses revealed that chrysoeriol inhibited the LPS-induced inductions of iNOS gene. Moreover, it is known that the activations of nuclear factor-kappaB (NF-kappaB) and activator protein 1 (AP-1) are crucial steps in the transcriptional activation of the iNOS gene. Here, we found that chrysoeriol selectively suppressed AP-1 activation, and that activation of AP-1 is likely to be essential for iNOS induction in LPS-treated macrophages. This presumed inhibitory effect on AP-1 activation by chrysoeriol may be associated with its potent NO blocking and anti-inflammatory effects. PMID:16228289

Choi, Doo-Youn; Lee, Jeong Yong; Kim, Mi-Ran; Woo, Eun-Rhan; Kim, Yoon Gyoon; Kang, Keon Wook

2005-12-01

374

Reduction of nitric oxide release from alveolar macrophages by a lipocortin peptide.  

PubMed Central

Nitric oxide (NO), produced by alveolar macrophages (AM) is used as a marker of respiratory tract inflammation. Lipocortin 1 (Lc-1) is an anti-inflammatory, glucocorticoid-inducible protein. The current aims were to determine whether (a) an Lc-1-derived peptide, Ac2-26, inhibited lipopolysaccharide (LPS)-induced NO release by primary AM in vitro and (b) the inhibitory action of dexamethasone was Lc-1-dependent. LPS treatment stimulated NO release from rat AM. Ac2-26 had little effect on unstimulated release, but suppressed LPS-stimulated release at concentrations > or =320 nM (320 nM, 10 +/- 3%; 3.2 microM, 15 +/- 3%; 32 microM, 27 +/- 4% NO inhibited, mean +/- SEM, n = 6). Inhibition by dexamethasone of NO release was unaffected by neutralizing anti-Lc-1 indicating that this action is Lc-1-independent in primary AM. Nevertheless inhibition of NO release by Ac2-26 (80 microM) was similar to that of 1 microM dexamethasone (Ac2-26, 40 +/- 6%; dexamethasone, 48 +/- 6% NO inhibited, mean +/- SEM, n = 6). PMID:9836495

Kamal, A M; Tetley, T D; Witherden, I R; Smith, S F

1998-01-01

375

Involvement of Syk kinase in TNF-induced nitric oxide production by airway epithelial cells  

SciTech Connect

We have recently found that Syk is widely expressed in lung epithelial cells (EC) and participates in {beta}1 integrin signaling. In this study, we assessed the role of Syk in regulation of NO production. Stimulation of human bronchial EC line HS-24 by TNF caused an increased expression of inducible nitric oxide synthase (iNOS). Inhibition of Syk using siRNA or piceatannol down-regulated the iNOS expression and reduced NO production. This effect occurred in EC simultaneously stimulated via {beta}1 integrins, suggesting that TNF and {beta}1 integrins provide co-stimulatory signals. Inhibition of Syk down-regulated TNF-induced p38 and p44/42 MAPK phosphorylation and nuclear translocation of p65 NF-{kappa}B. Thus, TNF-induced activation of pro-inflammatory signaling in EC leading to enhanced expression of iNOS and NO production was dependent on Syk. Syk-mediated signaling regulates NO production at least partly via activating the MAPK cascade. Understanding the role of Syk in airway EC may help in developing new therapeutic tools for inflammatory lung disorders.

Ulanova, Marina [Department of Medicine, University of Alberta, Edmonton, Alta. (Canada)]. E-mail: marina.ulanova@normed.ca; Marcet-Palacios, Marcelo [Department of Medicine, University of Alberta, Edmonton, Alta. (Canada); Munoz, Samira [Department of Medicine, University of Alberta, Edmonton, Alta. (Canada); Asfaha, Samuel [Department of Medicine, University of Alberta, Edmonton, Alta. (Canada); Kim, Moo-Kyung [University of Pennsylvania School of Medicine, Philadelphia, PA (United States); Schreiber, Alan D. [University of Pennsylvania School of Medicine, Philadelphia, PA (United States); Befus, A. Dean [Department of Medicine, University of Alberta, Edmonton, Alta. (Canada)

2006-12-15

376

Tetrahydrobiopterin Deficiency and Nitric Oxide Synthase Uncoupling Contribute to Atherosclerosis Induced by Disturbed Flow  

PubMed Central

Objective Tetrahydrobiopterin (BH4) is a critical cofactor for Nitric Oxide (NO) synthesis by NO synthase (NOS). Recently, we demonstrated that disturbed flow produced by partial carotid ligation decreases BH4 levels in vivo. We therefore aimed to determine whether atherosclerosis induced by disturbed flow is due to BH4 deficiency and NOS uncoupling and whether increasing BH4 would prevent endothelial dysfunction, plaque inflammation and atherosclerosis. Methods and Results We produced a region of disturbed flow in ApoE?/? mice using partial carotid ligation and fed these animals a high-fat diet. This caused eNOS uncoupling as characterized by increased vascular superoxide production, altered vascular reactivity and a change in eNOS migration on low-temperature gel. These perturbations were accompanied by severe atherosclerosis, infiltration of T cells and macrophages, and an increase in cytokine production. Treatment with BH4 recoupled NOS, decreased superoxide production, imporoved endothelium-dependent vasodilatation and virtually eliminated atherosclerosis. BH4 treatment also markedly reduced vascular inflammation and improved the cytokine milieu induced by disturbed flow. Conclusions Our results highlight a key role of BH4 deficiency and NOS uncoupling in atherosclerosis induced by disturbed flow, and provide insight into the effect of modulating vascular BH4 levels on atherosclerosis and inflammation at these sites of the circulation. PMID:21512164

Li, Li; Chen, Wei; Rezvan, Amir; Jo, Hanjoong; Harrison, David G.

2011-01-01

377

Nitric oxide-induced calcium release via ryanodine receptors regulates neuronal function  

PubMed Central

Mobilization of intracellular Ca2+ stores regulates a multitude of cellular functions, but the role of intracellular Ca2+ release via the ryanodine receptor (RyR) in the brain remains incompletely understood. We found that nitric oxide (NO) directly activates RyRs, which induce Ca2+ release from intracellular stores of central neurons, and thereby promote prolonged Ca2+ signalling in the brain. Reversible S-nitrosylation of type 1 RyR (RyR1) triggers this Ca2+ release. NO-induced Ca2+ release (NICR) is evoked by type 1 NO synthase-dependent NO production during neural firing, and is essential for cerebellar synaptic plasticity. NO production has also been implicated in pathological conditions including ischaemic brain injury, and our results suggest that NICR is involved in NO-induced neuronal cell death. These findings suggest that NICR via RyR1 plays a regulatory role in the physiological and pathophysiological functions of the brain. PMID:22036948

Kakizawa, Sho; Yamazawa, Toshiko; Chen, Yili; Ito, Akihiro; Murayama, Takashi; Oyamada, Hideto; Kurebayashi, Nagomi; Sato, Osamu; Watanabe, Masahiko; Mori, Nozomu; Oguchi, Katsuji; Sakurai, Takashi; Takeshima, Hiroshi; Saito, Nobuhito; Iino, Masamitsu

2012-01-01

378

The exudate from an arbuscular mycorrhizal fungus induces nitric oxide accumulation in Medicago truncatula roots.  

PubMed

Nitric oxide (NO) is a signaling molecule involved in plant responses to abiotic and biotic stresses. While there is evidence for NO accumulation during legume nodulation, almost no information exists for arbuscular mycorrhizas (AM). Here, we investigated the occurrence of NO in the early stages of Medicago truncatula-Gigaspora margarita interaction, focusing on the plant response to fungal diffusible molecules. NO was visualized in root organ cultures and seedlings by confocal microscopy using the specific probe 4,5-diaminofluorescein diacetate. Five-minute treatment with the fungal exudate was sufficient to induce significant NO accumulation. The specificity of this response to AM fungi was confirmed by the lack of response in the AM nonhost Arabidopsis thaliana and by analyzing mutants impaired in mycorrhizal capacities. NO buildup resulted to be partially dependent on DMI1, DMI2, and DMI3 functions within the so-called common symbiotic signaling pathway which is shared between AM and nodulation. Significantly, NO accumulation was not induced by the application of purified Nod factor, while lipopolysaccharides from Escherichia coli, known to elicit defense-related NO production in plants, induced a significantly different response pattern. A slight upregulation of a nitrate reductase (NR) gene and the reduction of NO accumulation when the enzyme is inhibited by tungstate suggest NR as a possible source of NO. Genetic and cellular evidence, therefore, suggests that NO accumulation is a novel component in the signaling pathway that leads to AM symbiosis. PMID:21744141

Calcagno, Cristina; Novero, Mara; Genre, Andrea; Bonfante, Paola; Lanfranco, Luisa

2012-05-01

379

COMPARISON OF INDUCIBLE NITRIC OXIDE SYNTHASE GENE EXPRESSION AND LUNG INFLAMMATION FOLLOWING INTRATRACHEAL INSTILLATION OF SILICA, COAL, CARBONYL IRON, OR TITANIUM DIOXIDE IN RATS  

Microsoft Academic Search

The pulmonary toxicity of the respirable dusts silica, coal, carbonyl iron, and titanium dioxide on alveolar macrophage (AM) and neutrophil (PMN) inducible nitric oxide syn thase (iNOS) gene expression and nitric oxide (NO) production was investigated. Rats were intratracheal^ instilled with 5 mg\\/100 g body weight of silica, coal, carbonyl iron, or titanium dioxide. The dust particles averaged less than

John A. Blackford Jr; William Jones; Richard D. Dey; Vincent Castranova

1997-01-01

380

The effect of free gallium and gallium in liposomes on cytokine and nitric oxide secretion from macrophage-like cells in vitro.  

PubMed

The aim of this study was to evaluate the effect of gallium nitrate, gallium-nitrilotriacetate (NTA) complex, and liposomal gallium-NTA on IL-6, TNF alpha, and nitric oxide (NO) release from activated macrophages. In addition, the expression of the inducible nitric oxide synthase (iNOS) was determined. Gallium inhibited dose-dependently the secretion of IL-6, TNF alpha, and NO from the LPS-induced macrophage-like RAW 264 cells. Encapsulation of gallium in negatively charged DSPG-liposomes increased its potency 10-50 times and 7-11 times compared to free gallium nitrate and gallium-NTA, respectively. Neither non-loaded liposomes nor NTA alone inhibited cytokine or NO secretion, demonstrating that the observed effects originated from gallium. Liposomal gallium-NTA inhibited the expression of iNOS by the macrophages, while other formulations of gallium had no effect. Thus, gallium, when delivered properly, suppresses macrophage functions by inhibiting the release of inflammatory mediators from the cells. PMID:8788232

Makkonen, N; Hirvonen, M R; Savolainen, K; Lapinjoki, S; Mönkkönen, J

1995-12-01

381

Absence of inducible nitric oxide synthase modulates early reperfusion-induced NF-kappaB and AP1 activation and enhances myocardial damage  

Microsoft Academic Search

The role of nitric oxide (NO) generated by the inducible NO synthase (iNOS) during myocardial ischemia and reperfusion is not understood. We investi- gated the role of iNOS during early reperfusion damage induced in genetically deficient iNOS (iNOS\\/) mice and wild-type littermates. In wild-type mice, ischemia (60 min) and reperfusion (60 min) induced an elevation in serum levels of creatine

BASILIA ZINGARELLI; PAUL W. HAKE; ZEQUAN YANG; ALVIN DENENBERG; HECTOR R. WONG

2002-01-01

382

Morphine-induced nitric oxide production in isolated, iris-ciliary bodies.  

PubMed

Considerable evidence suggests that the nitric oxide (NO)/cGMP signaling pathway plays an integral role in opioid receptor-mediated responses in the cardiovascular and immune systems. Previous studies in our laboratory and others have shown that nitric oxide (NO) plays a role in morphine-induced reduction of intraocular pressure (IOP) and pupil diameter (PD) in the New Zealand white (NZW) rabbit. The present study is designed to determine the effect of morphine on NO production in the isolated, iris-ciliary body (ICB), site of aqueous humor production, as this effect could be associated with morphine-stimulated changes in aqueous humor dynamics and iris function. ICBs obtained from normal NZW rabbits were utilized in these experiments. In some experiments, ICB samples were treated with morphine (1, 10 and 100 microM) for 1 h and later examined for changes in NO levels using a NO detection kit. In other experiments, tissue samples were pretreated with naloxone (non-selective opioid receptor antagonist), L-NAME (non-selective NO-synthase inhibitor) or GSH (sulfhydryl reagent) for 30 min, followed by treatment with morphine (10 muM). Morphine caused a concentration-dependent increase in the release of NO from ICBs. Levels of NO detected in the incubation medium of ICB samples increased from 1.49 +/- 0.19 (control) to 8.81 +/- 2.20 microM/mg protein (morphine-treated; 100 microM). Morphine-stimulated release of NO was significantly inhibited in tissues pretreated with 10 microM naloxone, L-NAME, or GSH. Results obtained from this study suggest that morphine stimulates NO release from the ICB through a mechanism that involves activation of NO-releasing opioid receptors. These results support the in vivo effects of morphine demonstrated in previous studies. PMID:19555685

Dortch-Carnes, Juanita; Randall, Karen Russell

2009-11-01

383

Catalytic Intermediates of Inducible Nitric-oxide Synthase Stabilized by the W188H Mutation*  

PubMed Central

Nitric-oxide synthase (NOS) catalyzes nitric oxide (NO) synthesis via a two-step process: l-arginine (l-Arg) ? N-hydroxy-l-arginine ? citrulline + NO. In the active site the heme is coordinated by a thiolate ligand, which accepts a H-bond from a nearby tryptophan residue, Trp-188. Mutation of Trp-188 to histidine in murine inducible NOS was shown to retard NO synthesis and allow for transient accumulation of a new intermediate with a Soret maximum at 420 nm during the l-Arg hydroxylation reaction (Tejero, J., Biswas, A., Wang, Z. Q., Page, R. C., Haque, M. M., Hemann, C., Zweier, J. L., Misra, S., and Stuehr, D. J. (2008) J. Biol. Chem. 283, 33498–33507). However, crystallographic data showed that the mutation did not perturb the overall structure of the enzyme. To understand how the proximal mutation affects the oxygen chemistry, we carried out biophysical studies of the W188H mutant. Our stopped-flow data showed that the 420-nm intermediate was not only populated during the l-Arg reaction but also during the N-hydroxy-l-arginine reaction. Spectroscopic data and structural analysis demonstrated that the 420-nm intermediate is a hydroxide-bound ferric heme species that is stabilized by an out-of-plane distortion of the heme macrocycle and a cation radical centered on the tetrahydrobiopterin cofactor. The current data add important new insights into the previously proposed catalytic mechanism of NOS (Li, D., Kabir, M., Stuehr, D. J., Rousseau, D. L., and Yeh, S. R. (2007) J. Am. Chem. Soc. 129, 6943–6951). PMID:23269673

Sabat, Joseph; Egawa, Tsuyoshi; Lu, Changyuan; Stuehr, Dennis J.; Gerfen, Gary J.; Rousseau, Denis L.; Yeh, Syun-Ru

2013-01-01

384

Inhibiting inducible nitric oxide synthase with rutin reduces renal ischemia/reperfusion injury  

PubMed Central

Background Nitric oxide (NO) seems to play an important role during renal ischemia/reperfusion (I/R) injury. We investigated whether rutin inhibits inducible nitric oxide synthase (iNOS) and reduces 3-nitrotyrosine (3-NT) formation in the kidneys of rats during I/R. Methods Wistar albino rats were nephrectomized unilaterally and, 2 weeks later, subjected to 45 minutes of left renal pedicle occlusion followed by 3 hours of reperfusion. We intraperitoneally administered L-N6-(1-iminoethyl)lysine (L-NIL; 3 mg/kg) for 30 minutes or rutin (1 g/kg) for 60 minutes before I/R. After reperfusion, kidney samples were taken for immunohistochemical analysis of iNOS and 3-NT. We measured plasma nitrite/nitrate and cyclic guanosine monophosphate (cGMP) to evaluate NO levels. Results Ischemia/reperfusion caused plasma cGMP to increase significantly. Similarly, plasma nitrite/nitrate was elevated in the I/R group compared with the control group. Histochemical staining was positive for iNOS and 3-NT in the I/R group. Pretreatment with L-NIL or rutin significantly mitigated the elevation of plasma cGMP and nitrite/nitrate. These changes in biochemical parameters were also associated with changes in immunohistochemical appearance. Pretreatment with L-NIL or rutin significantly decreased the incidence and severity of iNOS and 3-NT formation in the kidney tissues. Conclusion Our findings suggest that high activity of iNOS causes renal I/R injury, and that rutin exerts protective effects, probably by inhibiting iNOS. PMID:23187035

Korkmaz, Asli; Kolankaya, Dürdane

2013-01-01

385

Nitric oxide synthase promotes distension-induced tracheal venular leukocyte adherence.  

PubMed

The process of leukocyte recruitment to the airways in real time has not been extensively studied, yet airway inflammation persists as a major contributor to lung pathology. We showed previously in vivo, that neutrophils are recruited acutely to the large airways after periods of airway distension imposed by the application of positive end-expiratory pressure (PEEP). Given extensive literature implicating products of nitric oxide synthase (NOS) in lung injury after ventilatory over-distension, we questioned whether similar mechanisms exist in airway post-capillary venules. Yet, endothelial nitric oxide has been shown to be largely anti-inflammatory in other systemic venules. Using intravital microscopy to visualize post-capillary tracheal venules in anesthetized, ventilated mice, the number of adherent leukocytes was significantly decreased in eNOS-/- mice under baseline conditions (2±1 cell/60 min observation) vs wild type (WT) C57BL/6 mice (7±2 cells). After exposure to PEEP (8 cmH2O for 1 min; 5 times), adherent cells increased significantly (29±5 cells) in WT mice while eNOS-/- mice demonstrated a significantly decreased number of adherent cells (11±4 cells) after PEEP. A similar response was seen when thrombin was used as the pro-inflammatory stimulus. In addition, mouse tracheal venular endothelial cells studied in vitro after exposure to cyclic stretch (18% elongation) or thrombin both demonstrated increased p-selectin expression that was significantly attenuated by NG-nitro-L-arginine methyl ester, N-acetylcysteine amide (NACA) and excess BH4. In vivo treatment with the ROS inhibitor NACA or co-factor BH4 abolished completely the PEEP-induced leukocyte adherence. These results suggest that pro-inflammatory stimuli cause leukocyte recruitment to tracheal endothelium in part due to eNOS uncoupling. PMID:25181540

Moldobaeva, Aigul; Rentsendorj, Otgonchimeg; Jenkins, John; Wagner, Elizabeth M

2014-01-01

386

Nitric Oxide Synthase Promotes Distension-Induced Tracheal Venular Leukocyte Adherence  

PubMed Central

The process of leukocyte recruitment to the airways in real time has not been extensively studied, yet airway inflammation persists as a major contributor to lung pathology. We showed previously in vivo, that neutrophils are recruited acutely to the large airways after periods of airway distension imposed by the application of positive end-expiratory pressure (PEEP). Given extensive literature implicating products of nitric oxide synthase (NOS) in lung injury after ventilatory over-distension, we questioned whether similar mechanisms exist in airway post-capillary venules. Yet, endothelial nitric oxide has been shown to be largely anti-inflammatory in other systemic venules. Using intravital microscopy to visualize post-capillary tracheal venules in anesthetized, ventilated mice, the number of adherent leukocytes was significantly decreased in eNOS-/- mice under baseline conditions (2±1 cell/60 min observation) vs wild type (WT) C57BL/6 mice (7±2 cells). After exposure to PEEP (8 cmH2O for 1 min; 5 times), adherent cells increased significantly (29±5 cells) in WT mice while eNOS-/- mice demonstrated a significantly decreased number of adherent cells (11±4 cells) after PEEP. A similar response was seen when thrombin was used as the pro-inflammatory stimulus. In addition, mouse tracheal venular endothelial cells studied in vitro after exposure to cyclic stretch (18% elongation) or thrombin both demonstrated increased p-selectin expression that was significantly attenuated by NG-nitro-L-arginine methyl ester, N-acetylcysteine amide (NACA) and excess BH4. In vivo treatment with the ROS inhibitor NACA or co-factor BH4 abolished completely the PEEP-induced leukocyte adherence. These results suggest that pro-inflammatory stimuli cause leukocyte recruitment to tracheal endothelium in part due to eNOS uncoupling. PMID:25181540

Moldobaeva, Aigul; Rentsendorj, Otgonchimeg; Jenkins, John; Wagner, Elizabeth M.

2014-01-01

387

Fourier transform infrared spectroscopy study of ligand photodissociation and migration in inducible nitric oxide synthase  

PubMed Central

Inducible nitric oxide synthase (iNOS) is a homodimeric heme enzyme that catalyzes the formation of nitric oxide (NO) from dioxygen and L-arginine (L-Arg) in a two-step process. The produced NO can either diffuse out of the heme pocket into the surroundings or it can rebind to the heme iron and inhibit enzyme action. Here we have employed Fourier transform infrared (FTIR) pho