黄芪甲苷对阿霉素诱导大鼠心肌细胞凋亡的抑制作用

Inhibition effect of astragaloside on adriamycin-induced rat cardiomyocyte apoptosis

目的探讨黄芪甲苷对阿霉素(ADR)诱导大鼠心肌细胞凋亡的抑制作用。方法采用ADR 100ng/ml(ADR1组)、250ng/ml(ADR2组)和500ng/ml(ADR3组)作用于大鼠心肌细胞株H9C2,筛查出250ng/ml为最佳实验浓度,建立ADR诱导细胞凋亡模型。用黄芪甲苷10μg/ml(黄芪甲苷1组)、25μg/ml(黄芪甲苷2组)、50μg/ml(黄芪甲苷3组)作用于ADR诱导凋亡模型细胞,筛查出50μg/ml为最佳实验浓度。培养正常H9C2细胞为对照组。测定各组细胞的凋亡率、活性氧(ROS)、线粒体膜电位(ΔΨm)水平及心肌细胞Bax、Bcl-2 mRNA的表达水平。结果与对照组比较,ADR2组细胞凋亡率升高(P<0.01);与ADR2组比较,黄芪甲苷3组细胞凋亡率降低(P<0.05)。与对照组比较,ADR2组和黄芪甲苷3组ROS水平和ΔΨm降低比例增加(P<0.05);与ADR2组比较,黄芪甲苷3组ROS水平和ΔΨm降低比例减少(P<0.05)。与对照组比较,ADR2组Bax和Bcl-2mRNA表达升高(P<0.05),Bcl-2/Bax比值下降(P<0.05);与ADR2组比较,黄芪甲苷3组Bax mRNA表达降低(P<0.01),Bcl-2 mRNA和Bcl-2/Bax比值升高(P<0.01)。结论黄芪甲苷能够抑制ADR诱导的大鼠心肌细胞的凋亡,可能是通过降低细胞内ROS水平,提高ΔΨm,降低Bax而升高Bcl-2来发挥作用。

Objective To investigate the inhibition effect of astragaloside on adriamycin（ADR）-induced rat cardiomyocyte apoptosis.Methods The rat myocardial cell line H9C2 was treated with the ADR 100ng/ml（group ADR1）,250ng/ml（group ADR2）and 500ng/ml（group ADR3）.The ADR 250ng/ml was screened as the optimum experimental concentration and ADR-induced apoptosis model was established.The ADR-induced apoptosis model was treated with astragaloside 10μg/ml（group H1）,25μg/ml（group H2）and 50μg/ml（group H3）.The astragaloside 50μg/ml was screened as the optimum experimental concentration.The normal H9C2 cells were cultured as the controls（group C）.The cell apoptosis,reactive oxygen species（ROS）,mitochondrial membrane potential（ΔΨm）and the mRNA expressions of Bcl-2 and Bax were detected.Results The rate of cell apoptosis was higher in group ADR2 than that in group C（P〈0.01）,which was lower in group H3 than that in group ADR2（P〈0.05）.Compared to group C,the percentages of ROS reduction and ΔΨm were more in group ADR2（P〈0.05）.Compared to group C,the mRNA expressions of Bcl-2 and Bax were higher and the ratio of Bcl-2 to Bax was lower in group ADR2（P〈0.05）.The mRNA expression of Bax was lower and that of Bcl-2 and the ratio of Bcl-2 to Bax were higher in group H3 than those in group ADR2（P〈0.01）.Conclusion Astragaloside can inhibit the rat cardiomyocyte apoptosis induced by ADR,which may be related to the reduction of ROS,increase of ΔΨm,decrease of Bax expression and increase of Bcl-2.