【摘要】 目的探讨非对称性二甲基精氨酸(ADMA)上调大鼠主动脉诱导型一氧化氮合酶(iNOS)的表达是否是通过激活NF-κB实现的。方法 Wistar大鼠50只随机分为四组:①对照组(n=10):标准饲料喂养。②H组(n=12):高脂饲料喂养。③A+H组(n=14):予ADMA[0.2mg/kgd]灌胃,高脂饲料喂养。④P+A+H组(n=14):予吡咯烷二硫代氨基甲酸盐(pyrrolidine dithiocarbamate,PDTC)[40mg/kgd]腹腔注射、ADMA[0.2mg/kgd]灌胃、高脂饲料喂养。对照组、H组予等体积的生理盐水灌胃及腹腔注射、A+H组给予等体积的生理盐水腹腔注射。18周后麻醉大鼠、取主动脉。以实时荧光定量PCR和Westen blotting分别检测iNOS mRNA和蛋白表达,以电泳迁移率变动分析(EMSA)和增强化学发光法(ECL)检测NF-κB活性。结果①A+H组iNOS mRNA和蛋白表达量较对照组和H组增加(P<0.05),P+A+H组较A+H组iNOSmRNA和蛋白表达量降低(P<0.05)。②A+H组NF-κB活性较对照组和H组显著升高(P<0.05),P+A+H组NF-κB活性较A+H组明显降低(P<0.05)③相关分析:NF-κB活性与iNOS mRNA和蛋白表达量呈正相关(相关系数r分别为0.854、0.876,P<0.05)。结论 ADMA可能通过激活NF-κB途径上调iNOS表达,从而促进动脉粥样硬化的发生发展。更多还原

【Abstract】 Objective To investigate,whether the asymmetric dimethylarginine(ADMA) upregulated expression of iNOS in rat aortas is modulated by nuclear factor kappa B(NF-κB).Methods 50 Wistar rats were randomly divided into 4 groups:①Control group(n=10) fed with standard diet.②H group(n=12) fed with high fat diet.③ A+H group(n=14) administered intergastrically with ADMA [0.2mg/kgd],and fed with high fat diet.④ P+A+H group(n=14) given interperitoneal injection of pyrrolidine dithiocarbamate(PDTC) [40mg/kgd],administered intergastrically with ADMA [0.2mg/(kgd)],and fed with high fat diet.Both control and H groups were administered intergastrically and injected interperitoneally with the same volume of normal saline.The A+H group was injected interperitoneal,with the same volume of normal saline.18 weeks later,the rats were anesthetized,and aortas were gathered.The mRNA and protein expressions of iNOS were detected with real time fluorescent quantitative PCR(real-time PCR) and Westen Blotting respectively.The activity of NF-κB was detected with electrophorestic mobility shift assay(EMSA) and enhanced chemiluminescent(ECL) technique.Results ①The mRNA and protein expressions of iNOS in the A+H group were higher than those in control and H groups(P<0.05) decreased in P+A+H group than in the A+H group(P<0.05).②The NF-κB activity in the A+H group was extremely higher than in control and H groups(P<0.05);significantly lower in the P+A+H group than in the A+H group(P<0.05).③ The activity of NF-κB was positively correlated with the mRNA and protein expression of iNOS(P<0.05).Conclusion ADMA upregulates the iNOS expression through the NF-κB activity pathway in rat aortas,which might be one of the mechanisms of atherosclerosis.更多还原