抗纤益心方对去甲肾上腺素诱导H9c2心肌细胞凋亡及凋亡相关因子表达的影响

Effects of Kangxian Yixin Formula(抗纤益心方) on Norepinephrine-induced H9c2 Cardiomyocytes Apoptosis and Expression of Apoptosis-related Factors

目的探讨抗纤益心方对去甲肾上腺素(NE)诱导的H9c2心肌细胞凋亡的影响及可能作用机制。方法采用CCK-8法筛选抗纤益心方最佳给药浓度。大鼠H9c2心肌细胞常规培养,饥饿8 h后细胞进行分组,正常组未做处理,模型组加100μmol/L的NE 100μl,抗纤益心方组加入筛选最佳浓度抗纤益心方溶液2μl+100μmol/L的NE 100μl、卡托普利组加入2×10^-5mol/L卡托普利5μl+100μmol/L的NE 100μl,各组设3个复孔,作用24 h。流式细胞仪检测细胞凋亡率,并检测凋亡相关因子Bax、Bcl-2、Caspase-3 mRNA和蛋白的表达。结果最终筛选以0.25 mg/ml的抗纤益心方为干预浓度进行实验。与正常组比较,模型组细胞凋亡率升高,Bax、Caspase-3 mRNA和蛋白表达升高,Bcl-2 mRNA和蛋白表达降低(P<0.01)。与模型组比较,抗纤益心方组和卡托普利组细胞凋亡率降低,Bax、Caspase-3 mRNA和蛋白表达降低,Bcl-2 mRNA和蛋白表达升高(P<0.05或P<0.01)。结论抗纤益心方可抑制NE诱导的H9c2心肌细胞凋亡,其作用机制可能与调节凋亡相关因子Bax、Bcl-2、Caspase-3的表达有关。

Objective To investigate the effects of Kangxian Yixin Formula(抗纤益心方)on norepinephrine(NE)-induced apoptosis of H9c2 cardiomyocytes and its possible mechanism.Methods The optimal concentration of Kangxian Yixin Formula was screened by CCK-8 method.Rat H9c2 cardiomyocytes were cultured routinely,and cells were grouped after starvation for 8 h.The normal group was not treated.The model group was supplemented with 100μmol/L NE 100μl.The Kangxian Yixin Formula was added to the optimal concentration of Kangxian Yixin Formula solution 2μl+100μmol/L NE 100μl,captopril group 2×10-5mol/L captopril 5μl+100μmol/L NE 100μl,each group had 3 duplicate wells for 24 h.The apoptosis rate was detected by flow cytometry,and the expressions of apoptosis-related factors Bax,Bcl-2 and Caspase-3 mRNA and protein were detected.Results The final screening was performed at an intervention concentration of 0.25 mg/ml Kangxian Yixin Formula.Compared with the normal group,the apoptosis rate of the model group increased,the expression of Bax and Caspase-3 mRNA and protein increased,and the expression of Bcl-2 mRNA and protein decreased(P<0.01).Compared with the model group,the apoptosis rate of the Kangxian Yixin Formula group and the captopril group decreased,the expression of Bax and Caspase-3 mRNA and protein decreased,and the expression of Bcl-2 mRNA and protein increased(P<0.05 or P<0.01).Conclusion Kangxian Yixin Formula can inhibit NE-induced H9c2 cardiomyocyte apoptosis,and its mechanism may be related to the regulation of apoptosis-related factors Bax,Bcl-2 and Caspase-3.