Title: Asymmetric dimethylarginine induces inducible nitric oxide synthase expression via NF-κB pathway in rat peritoneal macrophages
Abstract: Objective:To investigate whether asymmetric dimethylarginine(ADMA)upregulated the expression of inducible nitric oxide synthase(iNOS)in rat peritoneal macrophages is modulated by nuclear factor kappa B (NF-κB)pathway.Method:Primary cultured Wistar rat peritoneal macrophages were divided into four groups:① Control group:macrophages incubated in DMEM media with 10%fetal bovine serum(FBS)without any intervention;②oxLDL group:macrophages incubated in DMEM media with 10% FBS and oxidized low density lipoprotein(oxLDL,50mg/L);③A+O group:macrophages incubated in DMEM media with 10% FBS,ADMA (15 μmol/L)and oxLDL(50mg/L);④P+A+O group:macrophages incubated in DMEM media with 10% FBS, PDTC(25μmol/L),ADMA(15μmol/L)and oxLDL(50mg/L).The mRNA and protein expression of iNOS were detected by real time fluorescent quantitative PCR(real-time PCR)and Westen Blotting respectively.The activity of NF-κB was detected by electrophoretic mobility shift assay(EMSA)and enhanced chemiluminescent (ECL)technique.Result:①Compared with control group,the mRNA and protein expression of iNOS were significantly increased in oxLDL group and A+O group,especially in A+O group(P0.05);the mRNA and protein expression of iNOS in P+A+O group were significantly decreased than those in A+O group(P0.05).②The NF-κB activity in both A+O group and oxLDL group was extremely higher than that in control group,especially in A+O group(P0.05);and the NF-κB activity in P+A+O group was significantly lower than that in A+O group(P0.05).③Correlative analysis:The activity of NF-κB was positively correlated with the mRNA and protein expression of iNOS(P0.05).Conclusion:ADMA upregulates the iNOS expression through the NF-κB activity pathway in rat peritoneal macrophages,which might be one of mechanisms of foam cell formation and atherosclerosis.
Publication Year: 2010
Publication Date: 2010-01-01
Language: en
Type: article
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