黄芪总皂苷对三氧化二砷诱导H9c2细胞损伤的保护作用

Protective effect of total saponins of astragalus on H9c2 cell damage induced by arsenic trioxide

为探讨黄芪总皂苷(ASTs)对三氧化二砷(ATO)诱导H9c2细胞损伤的保护作用及其机制,采用6μg·mL^(-1) ATO孵育12 h建立H9c2细胞损伤模型.实验设空白组、模型组、ASTs低/中/高浓度组,ASTs组预孵育12 h,弃去含药培养基,继续用6μg·mL^(-1) ATO孵育12 h,终止实验.通过CCK-8法检测细胞存活率,Hoechst 33342染色法观察细胞凋亡程度,流式细胞术定量测定细胞凋亡率,RT-qPCR测定Bcl-2、Bax、Caspase-3、Caspase-9表达水平,Western Blot测其蛋白.结果表明:ATO能够显著降低心肌细胞存活率、增加凋亡率(P<0.05);与ATO模型组比较,ASTs各组能显著提高细胞存活率,降低细胞凋亡率(P<0.05),降低Caspase-3、Caspase-9、Bax mRNA表达水平,增加Bcl-2 mRNA表达水平(P<0.05),显著降低细胞中Caspase-3、Caspase-9蛋白表达水平(P<0.05),升高Bcl-2/Bax比值(P<0.05).故ASTs对ATO诱导H9c2细胞凋亡具有保护作用,其机理可能与抑制细胞凋亡有关.

To investigate the protective mechanism of total saponins of astragalus(ASTs)on H9c2 cell injury induced by arsenic trioxide(ATO),the H9c2 cells injury model was established by incubating with 6μg·mL^(-1) ATO for 12 hours,and the blank group,model group,ASTs low,medium and high concentration group were set up in the experiment.Each ASTs group was pre-incubated for 12 hours,then the drug-containing medium was discarded,and the experiment was terminated with 6μg·mL^(-1) ATO for 12 hours.Cell viability was measured by the CCK-8 method,and the degree of apoptosis was observed by the Hoechst 33342 staining method.The quantitative apoptosis rate was determined by flow cytometry,and the expression levels of apoptosis-related genes Bcl-2,Bax,Caspase-3,Caspase-9 were examined by RT-qPCR and these proteins were tested by Western Blot.The results indicated that ATO could significantly reduce the survival rate of cardiomyocytes and increase the rate of apoptosis(P<0.05);compared with ATO group,each ASTs group could significantly enhance the survival rate of cells,reduce the rate of apoptosis(P<0.05),decrease Caspase-3,Caspase-9,Bax mRNA expression levels,increase Bcl-2 mRNA expression level(P<0.05),decrease Caspase-3 and Caspase-9 protein levels in cells(P<0.05),and increase Bcl-2/Bax(P<0.05).Therefore,ASTs has a protective effect on ATO-induced H9c2 cells apoptosis,and the mechanism may be related to the inhibition of apoptosis.