Experimental Study On Effect Of AS Ⅳ On ADR-Enduced H9C2 Cardiomyocytes Apoptosis In Rats

Objective:Through observing the effect of myocardial cell apoptosis rate, apoptosis gene Bax, bcl-2 mRNA expression, apoptosis related factor of active oxygen free radicals (ROS) and mitochondrial membrane potential (ΔΨm) with adriamycin induced rat H9c2 myocardial cells,to explore the possible mechanism of astragaloside IV on H9c2 myocardial cell apoptosis induced by adriamycin.Methods:Different concentrations (100ng/ml、250ng/ml、500ng/ml)of adriamycin were used to screen out the best experimental concentration, and the model of adriamycin induced cell apoptosis was established by using H9c2 cells.Low, middle and high dose groups (10ug/ml, 25ug/ml,50ug/ml) were used in the model cell, and the concentration of astragaloside IV was selected in the experiment.The experimental cells were divided randomly into astragaloside IV group, model group and blank control group.After administration, annexin V/PI staining and flow cytometry was used to detect the cell apoptosis percentage, RT-PCR to detect the Bax, Bcl-2 mRNA expression level, flow cytometry to the level of oxygen free radicals (ROS) and mitochondrial membrane potential (ΔΨm).Results:1.Percentage of myocardial cell apoptosis:Compared with the normal control group (39.72%), the percentage of myocardial cell apoptosis in model group (91.06%) was significantly increased;Compared with the model group, the percentage of apoptosis in myocardial cells (60.01%) was significantly decreased.2. Expression of Myocardial apoptosis gene Bax,Bcl-2 mRNA:Bax mRNA:Compared with the normal control group (0.31+1.36), the expression of model group (3.01+1.43) was significantly higher;Compared with the model group, the expression of the AS IV group (1.86+0.32) was significantly lower than that of the model group(P<0.05).Bcl-2 mRNA:Compared with the normal control group (0.22+0.76), the expression of model group (1.20+0.38) was increased;Compared with the model group, the expression of AS Ⅳ group (1.81+0.25) was significantly increased(P<0.01).3.Percentage of ROS increased in cardiac myocytes:Compared with the normal control group (16.09%), the percentage of ROS increased significantly in model group (63.35%); Compared with the model group, the percentage of ROS increased significantly in the AS Ⅳ group (28.81%).4.The percentage of ΔΨm reduced in myocardial cells: compared with the normal control group(26.40%), model group (51.13%) was increased significantly;Compared with the model group, AS Ⅳ group (35.46%) was significantly reduced.Conclusion:1. Astragaloside Ⅳ can decrease the apoptosis rate of H9C2 induced by adriamycin in rats; 2. Astragaloside Ⅳ can reduce the expression level of Bax mRNA, and increase the expression level of Bcl-2 mRNA in H9C2 rat cardiomyocytes induced by adriamycin;3.Astragaloside IV can decrease the level of reactive oxygen free radicals and enhance the level of mitochondrial membrane potential in H9C2 rat cardiomyocytes induced by adriamycin; 4.The mechanism of Astragaloside IV on H9c2 myocardial cell apoptosis in rats may be associated with regulation of apoptosis gene Bax、bcl-2 mRNA expression and regulation of apoptotic factors ROS、ΔΨm level.