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目的:观察二甲双胍对原代心肌细胞凋亡的影响。方法:利用乳鼠心室肌组织培养出大鼠原代心肌细胞。实验分为正常心肌细胞组、心肌细胞+过氧化氢(100μmol/L)损伤组、心肌细胞+过氧化氢(100μmol/L)+二甲双胍组(100μmol/L)。TUNEL试剂盒检测心肌细胞凋亡。Western blotting方法检测各组心肌细胞中凋亡相关蛋白3(Caspase-3)蛋白水平表达。结果:正常心肌细胞组中,心肌细胞凋亡指数为(5.99±0.61)%,活性氧使心肌细胞的凋亡率明显增加为(33.45±1.78)%,二甲双胍可明显抑制细胞凋亡,凋亡率减为(7.89±1.12)%,均差异有统计学意义(均P<0.05)。Western blotting结果显示与正常心肌细胞组比较,活性氧作用的心肌细胞内Caspase-3表达水平明显升高,二甲双胍组可显著抑制活性氧损伤的心肌细胞内Caspase-3蛋白表达(P<0.05)。结论:二甲双胍能明显减少活性氧造成心肌细胞的凋亡,这种抗心肌细胞凋亡的作用和抑制Caspase-3蛋白表达有关。
Objective: To observe the effect of metformin on primary cardiomyocyte apoptosis. Methods: Rat primary cardiomyocytes were cultured using neonatal rat ventricular myocytes. The experiment was divided into normal cardiomyocyte group, cardiomyocyte + hydrogen peroxide (100μmol / L) injury group, cardiomyocyte + hydrogen peroxide (100μmol / L) and metformin group (100μmol / L). TUNEL kit was used to detect cardiomyocyte apoptosis. Western blotting was used to detect the protein expression of apoptosis related protein 3 (Caspase-3) in each group. Results: The apoptotic index of cardiomyocytes was (5.99 ± 0.61)% in normal cardiomyocytes group and (33.45 ± 1.78)% in reactive oxygen species. Metformin significantly inhibited the apoptosis and apoptosis of cardiomyocytes The rate was reduced to (7.89 ± 1.12)%, both with statistical significance (all P <0.05). Western blotting showed that compared with normal cardiomyocytes, Caspase-3 expression was significantly increased in ROS treated cardiomyocytes, and Metformin significantly inhibited Caspase-3 protein expression (P <0.05). CONCLUSION: Metformin can significantly reduce the apoptosis of cardiomyocytes induced by reactive oxygen species (ROS), which is related to the inhibition of Caspase-3 protein expression.