摘要
目的:探讨同型半胱氨酸硫内酯(homocysteine thiolactone,HcyT)对人脐静脉内皮细胞凋亡及caspase3基因表达的影响及抗氧化剂干预后上述指标的改变。方法:一定浓度HcyT及抗氧化剂作用细胞后,碘化吡啶染色流式细胞仪、琼脂糖凝胶电泳检测细胞凋亡,RT-PCR和Western blot免疫印记法测定caspase3mRNA和蛋白表达。结果:HcyT以剂量依赖性方式诱导内皮细胞凋亡且caspase3mRNA及蛋白表达随其作用浓度的增加而升高(P<0.05)。1mmol/L HcyT浓度时,应用抗氧化剂干预,N-乙酰胱氨酸、维生素E及核转录因子抑制剂吡咯烷二硫氨基甲酸使凋亡细胞百分率从(23.16±4.27)%分别降至(1.95±0.23)%、(11.01±2.62)%、(11.46±1.81)%(P<0.05)。结论:HcyT诱导内皮细胞凋亡过程中,caspase3发挥重要作用。抗氧化剂可能通过抑制氧化损伤,下调caspase3表达,对内皮细胞凋亡发挥防治作用。
Objective：To investigate the effects of homocysteine thiolactone （HcyT） on apoptosis and caspase3 expression in human umbilical vein endothelial cell （HUVEC） and study the role of some antioxidants （ N-acetylcysteine NAC, vitamin E VitE, and pyrrolidine dithiocarbamate PDTC） in the reduction of HcyT -induced apoptosis. Methods： Flow cytometer and agarose electrophoresis of DNA fragmentation were applied to detect and observe cell aoptosis, mRNA and protein expression of caspase3 were determined by RT-PCR and Western blotting. Results：HcyT could induce cell apoptosis in a concentration-dependent manner. The mRNA and protein expression of caspase3 were up-regulated in concordance with the increased HcyT concentration. NAC, VitE, or PDTC reduced HcyT-indued apoptosis, concurrently down-regulated caspase3 expression. Conclusion： HcyT exerted its genotoxic effects on HUVEC through an apoptotic pathway, in which caspase3 might play an important role. The strong antioxidant character may largely account for effects of NAC, VitE or PDTC on apoptosis reduction induced by HcyT.
出处
《军医进修学院学报》
CAS
北大核心
2006年第3期178-180,共3页
Academic Journal of Pla Postgraduate Medical School
