丹参饮提取物A通过PI3K-Akt信号通路抗细胞凋亡机制的研究

Study on Anti-apoptotic Mechanism of Danshen Yin Extract A through the PI3K-Akt Signaling pathway

目的探讨丹参饮提取物A(Danshen Yin's extracts A,DSYEA)抗心肌缺血大鼠心肌细胞凋亡的作用机制。方法采用大鼠冠脉结扎手术制备心肌缺血大鼠模型。取70只大鼠随机分为正常组、模型组、复方丹参滴丸组(含生药量78.7 g·kg^(-1))、尼可地尔组(含生药量2.91 g·kg^(-1))及丹参饮提取物A(DSYEA)低、中、高剂量组(含生药量3.88,7.76,15.52 g·kg^(-1)),正常组和模型组灌胃等量蒸馏水。给药30 d后,TUNEL法测定细胞凋亡指数;免疫组化法检测Bax/Bcl-2蛋白的表达;Westen Blot法检测Caspase-3、P-AKT、p-GSK-3β蛋白的表达。结果与模型组比较,DSYEA中、高剂量组可下调心肌细胞凋亡指数及Bax蛋白、Caspase-3蛋白的表达水平(P<0.05,P<0.01),促进Bcl-2蛋白、P-GSK-3β、P-Akt蛋白的表达(P<0.05,P<0.01)。结论DSYEA可降低心肌缺血大鼠心肌细胞的凋亡,其机制可能与通过PI3K-AKt信号通路调控凋亡蛋白有关。

Objective To explore the anti-apoptosis mechanism of Danshen Yin extract A （DSYEA） in myocardial ischemia in rats. Methods The rats of myocardial ischemia were developed by ligation of the coronary artery. Seventy rats were randomized into normal group, model group, Compound clans he n dripping pill group （78.7 g·kg^-1） , nieorandil group （2.91g·kg^-1） , and low-, medium-, high-close DSYEA groups （3.88, 7.76, 15.52g·kg^-1）. The normal group and model group were intragastric administered with water. After treatment for 30 days, apoptosis index is determined by TUNEL; the expression of Bax/Bcl-2 pathway proteins was detected by immunohistochemistry, and the expression levels of Caspase-3, p-AKT, p-GSK-3 [3 were detected by Western Blot. Results Compared with model group, medium- and high-dose DSYEA groups reduced myocardial apoptosis and the Bax, Caspase-3 expression levels （P 〈 0.05, P 〈 0.01） , enhanced the Bcl-2, p-GSK-3β, p-Akt expression levels （P 〈 0.05, P 〈 0.01）. Conclusion DSYEA might inhibit the cardiomyocyte apoptosis, and the mechanism might be related to regulation of the apoptosis proteins through the PI3K-Akt signaling pathway.