The effect of Shenfu on ischemia-reperfusion induced cardiomyocyte apoptosis in rats

Objective:To observe the effect of Shenfu injection on the ultrastructure and apoptosis of myocytes in rats with myocardial ischemia/reperfusion injuries,and to explore the protective mechanisms of Shenfu injection on myocytes.Methods:The myocardial ischemia /reperfusion models is established by ligating the left anterior descending coronary arteries(LAD) of rats for 30 min,and then reperfused for 10 min.A total of 46 rats were divided into 5 groups,(Ⅰ)Sham-operated group;(Ⅱ) myocardial ischemia 30 min + saline;(Ⅲ) myocardial ischemia 30 min/reperfusion 10 min + saline;(Ⅳ) treatment group myocardial ischemia 30 min + Shenfu injection;(Ⅴ) treatment group myocardial ischemia 30 min/reperfusion 10 min + Shenfu injection.Each group was collected the ventricular muscle tissue from the same part of the heart.Transmission electron microscopy was applied to observe the changes in myocardial ultrastructure,at 25,000-fold magnification,the changes of myocardial mitochondrial ultrastructure were analyzed by CIMAS image system.Myocardial cell apoptosis was determined by TUNEL method,the expression of Bcl-2,Bax protein in myocardial cells was detected with immunohistochemistry technique.The expression of Bcl-2,Bax protein and cellapoptosis were ana-lyzed by pathological image analysis system.Results:Compared to control group(Ⅰ),in Shenfu-treated group(Ⅳ、Ⅴ),the expression of Bcl-2 protein was increased significantly(P 0.01),and a slight increase of Bax protein expression in Ⅳand significantly reduced(P 0.05,P 0.05).Shenfu-treated group significantly reduced myocardial apoptosis(P 0.05);Myocardial cell ultrastructure shows inⅤtreatment group had significantly reduced the damage;Myocardial ultrastructure stereologic analysis showed that the control group compared with the sham-operated group had a significant change in mitochondria(P 0.01),Shenfu-treated group(Ⅳ、 Ⅴ) compared with the sham-operated group of mitochondrial degree of change was no significant difference(P 0.05).Conclusion:Shenfu injection could inhibit myocardial cell apoptosis in rats with myocardial ischemia/reperfusion and its mechanism may be through up-regulating Bcl-2 protein expression and down-regulating Bax protein expression.