白藜芦醇对大鼠缺氧心肌细胞凋亡的保护作用及对SIRT1表达的影响

Effects of resveratrol on cardiomyocyte apoptosis induced by anoxia and the expression of SIRT1

目的研究白藜芦醇(Res)对大鼠缺氧心肌细胞凋亡的保护作用,以及对SIRT1表达的影响。方法选择褐鼠心肌细胞株H9c2培养48 h后随机分为5组:Res干预组,Res对照组,尼克酰胺(Nam)干预组,Nam对照组和正常对照组。24 h后终止培养,RT-PCR法检测SIRT1 mRNA表达水平的变化;免疫细胞化学法和Western blot法检测sIRT1蛋白表达水平的变化;TUNEL法及流式细胞仪PI染色法分析细胞凋亡。结果与Res对照组比较,Res干预组SIRT1 mRNA表达水平升高,蛋白表达增加;与Nam对照组比较,Nam干预组SIRT1 mRNA水平与蛋白表达减少,差异均有统计学意义(P<0.05,P<0.01)。与Res对照组比较,Res干预组细胞凋亡率降低,与Nam对照组比较,Nam干预组细胞凋亡率升高(P<0.01),正常对照组细胞凋亡率最低。结论 Res能减少缺氧诱导的心肌细胞凋亡,可能是通过上调SIRT1的表达发挥作用。

Objective To study the protective effect of resveratrol against cardiomyocyte apoptosis induced by hypoxia and the expression of SIRT1. Methods Cardiomyocytes of strain H9c2 were randomly divided into five groups after culturing for 48 h：20μmol/L resveratrol（Res）group,Res control group, 40 mmol/L nicotinamide （Nam） group, Nam control group, and normal control group. Twenty-four h later,the SIRT1 mRNA was examined by the RT-PCR. Immunocytochemical assay and Western blot were performed to examine the SIRT1 protein expression. Cell apoptosis was examined by terminal deoxynucleotidyltransferase-mediated dUTP nick-end lableling （TUNEL） and flow cytometry（FCM）. Results Compared with Res control group,Res promoted the expression of SIRT1 mRNA and protein （P〈0.05 ,P〈0.01）. However,the expression of SIRT1 mRNA and protein was lower in Nam group than in Nam control group （P〈0.05,P〈0.01）. The rate of cell apoptosis,which was 0.37% in the normal control group,was much lower in Res group （4.76%） than in Res control group （11.60%）. However,it was much higher in the Nam group （22.73%） than in the Nam control group （13.42%）. The same changes of cell apoptosis were observed by FCM analysis. Conclusion Resveratrol protects the cardiomyocytes from anoxia-induced apoptosis,which may be implemented by promoting the expression of SIRT1.