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目的研究白藜芦醇-Sirt1-Foxo1调控通路对缺氧心肌细胞的保护作用。方法将大鼠心肌细胞株H9C2培养48h后,随机分为5组,即20μmol/L白藜芦醇(resveratro,lRes)干预组、二甲基亚砜(dimethylsulfoxide,DMSO)对照组、40mmol/L尼克酰胺(nicotinamide,Nam)干预组、Nam空白对照组及正常对照组。培养24h后终止培养,用RT-PCR法和免疫细胞化学染色法检测Sirt1、Foxo1、p27、BimmRNA及其蛋白表达水平的变化;用TUNEL法及流式细胞仪(flowcytometer,FCM)分析细胞凋亡和细胞周期。结果20μmol/LRes干预组可使Sirt1mRNA及SIRT1蛋白表达的水平增加。Sirt1可通过抑制Foxo1mRNA的转录活性而调节其下游基因p27及Bim的表达。SIRT1的高表达可使细胞周期延长,细胞凋亡减少。结论Res干预可使SIRT1表达增加。Sirt1可能通过对Foxo1及其下游基因Bim、p27的调控,延长心肌细胞的细胞周期,减少其凋亡。Protectiveeffectofresveratrol-Sirt1-Foxo1pathwayonanoxiccardiocytes
Objective To study the protective effect of resveratrol-Sirt1-Foxo1 pathway on hypoxic cardiomyocytes. Methods H9C2 cells were cultured in vitro for 48 h. The rats were randomly divided into 5 groups: 20 μmol / L resveratrol (lRes) group, dimethylsulfoxide (DMSO) control group, 40 mmol / L Nicotinamide (Nam) intervention group, Nam blank control group and normal control group. After cultured for 24h, culture was terminated, and the changes of Sirt1, Foxo1, p27, BimmRNA and their protein expression were detected by RT-PCR and immunocytochemical staining; apoptosis was analyzed by TUNEL and flow cytometry (FCM) And cell cycle. Results The levels of Sirt1 mRNA and SIRT1 protein were increased in 20μmol / LRes intervention group. Sirt1 regulates the expression of its downstream genes p27 and Bim by inhibiting the transcriptional activity of Foxo1 mRNA. The high expression of SIRT1 can prolong cell cycle and reduce apoptosis. Conclusion Res intervention can increase SIRT1 expression. Sirt1 may be through the regulation of Foxo1 and its downstream genes Bim, p27, to extend the cell cycle of cardiac myocytes and reduce their apoptosis. Protective effect of resveratrol-Sirt1-Foxo1pathway anoxic cardiocytes