益气温阳活血利水方对AngⅡ诱导H9c2心肌细胞线粒体凋亡途径的影响

Effect of Yiqi Wenyang Huoxue Lishui formula on mitochondria apoptosis pathway of H9c2 myocardial cells induced by AngⅡ

目的观察益气温阳活血利水方含药血清对AngII诱导H9c2心肌细胞线粒体凋亡途径相关Bax、Bcl-2、Caspase-9基因及蛋白的表达变化,探讨益气温阳活血利水方对AngII诱导H9c2心肌细胞凋亡的保护作用的机制。方法AngII诱导H9c2心肌细胞凋亡模型,采用血清药理学方法制备益气温阳活血利水方含药血清并进行干预,使用qRT-PCR、Western Blot、免疫组化方法检测Caspase-9 mRNA和蛋白及Bax、Bcl-2蛋白表达变化。结果益气温阳活血利水方含药血清可提高Bcl-2蛋白表达,降低Bax蛋白表达,下调Caspase-9基因及蛋白的表达水平,且含药血清高剂量组改善作用最好。结论益气温阳活血利水方含药血清对AngII诱导H9c2心肌细胞凋亡的保护作用是通过调节线粒体凋亡途径相关Bcl-2、Bax基因蛋白的表达,从而抑制Caspase-9的表达实现的。

Objective To observe the effect of drug-containing serum of Yiqi Wenyang Huoxue Lishui formula on the genes and proteins expression levels of Bax,Bcl-2 and Caspase-9 related to mitochondria apoptosis pathway of H9c2 myocardial cells induced by AngⅡ.And to explore the protective mechanism of Yiqi Wenyang Huoxue Lishui formula on H9c2 myocardial cells apoptosis induced by AngⅡ.Methods The model of H9c2 myocardial cells apoptosis induced by AngⅡ was established.Then the drug-containing serum of Yiqi Wenyang Huoxue Lishui formula was prepared with serum pharmacology method and the model was intervened with it.The methods of qRT-PCR,Western Blot and immunohistochemistry were used to detect the genes and proteins expression levels of Caspase-9 and the proteins expression levels of Bax、Bcl-2.Results The drug-containing serum of Yiqi Wenyang Huoxue Lishui formula could increase the protein expression level of Bcl-2,decreasing the protein expression level of Bax,and down-regulating the gene and protein expression level of Caspase-9.And the improvement effect was best in the high dose of drug-containing serum group.Conclusion Drug-containing serum of Yiqi Wenyang Huoxue Lishui formula have a protective effect on H9c2 myocardial cells induced by AngⅡ,which is achieved by regulating the gene and protein expression levels of Bcl-2 and Bax related to the mitochondrial apoptosis pathway,then inhibiting the expression of Caspase-9.