益气活血方对缺氧H9c2心肌细胞凋亡的影响

Effect of Yiqi Huoxue Decoction on Apoptosis of Hypoxic H9c2 Cardiomyocytes

目的:观察益气活血方对过氧化氢(H2O2)诱导心肌细胞凋亡的保护作用。方法:培养H9c2心肌细胞,设为空白对照组、模型组(200μmol·L^(-1))、益气活血方组(0.75 g·L^(-1))、辛伐他汀组(5μmol·L^(-1)),药物预干预16 h后,除空白对照组外,其余组细胞用H2O2处理6 h。MTT法测定细胞活力;AnnexinV-FITC双染法检测细胞凋亡水平;Western Blot法检测Bcl-2、Bax及Caspase-3蛋白表达。结果:与空白对照组比较,模型组细胞活力明显降低(P<0.05),细胞凋亡明显增加(P<0.05),明显降低Bcl-2/Bax水平,明显下调Caspase-3表达(P<0.05);与模型组比较,益气活血方组、辛伐他汀组细胞活力明显增高、细胞凋亡率明显减少,明显升高Bcl-2/Bax水平,明显上调Caspase-3表达(P<0.05)。结论:益气活血方可明显抑制H2O2诱导的H9c2心肌细胞活力,升高Bcl-2/Bax水平,下调Caspase-3蛋白表达,可能与抑制细胞凋亡有关。

Objective: To observe the protective effect of Yiqi Huoxue Decoction on cardiomyocyte apoptosis induced by hydrogen peroxide(H2O2).Methods: H9 c2 cardiomyocytes were cultured and divided into blank control group, model group(200 μmol·L^(-1)),Yiqi Huoxue Decoction group(0.75 g·L^(-1)) and simvastatin group(5 μmol·L^(-1)).After 16 hours of drug pre-intervention, the cells of other groups were treated with H2O2 for 6 hours except the blank control group.Cell viability was measured by MTT assay;Annexin V-FITC double staining was used to detect the level of apoptosis;the expressions of Bcl-2,Bax and Caspase-3 proteins were detected by Western Blot.Results: Compared with the blank control group, the cell viability of the model group decreased significantly(P<0.05),the apoptosis increased significantly(P<0.05),the level of Bcl-2/Bax decreased significantly, and the expression of Caspase-3 decreased significantly(P<0.05).Compared with the model group, the cell viability, apoptosis rate, Bcl-2/Bax level and Caspase-3 expression were significantly increased in Yiqi Huoxue Decoction group and simvastatin group(P<0.05).Conclusion: Yiqi Huoxue Decoction can significantly inhibit the apoptosis of H9 c2 cardiomyocytes induced by H2O2,increase the level of Bcl-2/Bax and down regulate the expression of caspase-3 protein, which may be related to apoptosis.