白藜芦醇通过激活PI3K/Akt通路抑制心肌梗死大鼠细胞线粒体凋亡的作用研究

Study on the effect of Resveratrol in inhibiting mitochondrial apoptosis of rats with myocardial infarction by activating PI3K/Akt pathway

目的:探究白藜芦醇通过激活PI3K/Akt通路抑制心肌梗死大鼠细胞线粒体凋亡的作用机制。方法:选择45只成年雄性SD大鼠,将其随机分为对照组、模型组和白藜芦醇组,每组15只。将模型组和白藜芦醇组建立缺血-再灌注(I/R)模型,对白藜芦醇组大鼠舌静脉注射白藜芦醇进行干预,检测和比较大鼠心肌细胞损伤情况和凋亡情况。分别使用蛋白免疫印迹法(Western blot)和实时荧光定量聚合酶链反应(qRT-PCR)法检测磷脂酰肌醇3激酶-蛋白激酶B(PI3K/Akt)通路和线粒体凋亡基因表达水平。结果:模型组大鼠血清肌钙蛋白T(cTnT)和肌酸激酶同工酶(CK-MB)水平显著升高,白藜芦醇组大鼠血清cTnT和CK-MB水平显著低于模型组,其差异有统计学意义(t=27.129,t=14.971;P<0.05)。模型组大鼠的心肌细胞凋亡率显著升高,白藜芦醇组大鼠心肌细胞凋亡率显著低于模型组,其差异有统计学意义(t=12.687,P<0.05)。建模后心肌细胞中的PI3K/Akt通路被显著抑制,而白藜芦醇可显著促进PI3K/Akt通路活化。建模后大鼠心肌细胞B淋巴细胞瘤-2(Bcl-2)mRNA水平下调而细胞死亡调解子(BIM)、Bcl-2相关蛋白X(BAX)、半胱天冬酶9(Caspase9)、半胱天冬酶3(Caspase3)mRNA水平上调,白藜芦醇可显著抑制BIM、BAX、Caspase9以及Caspase3基因表达并促进Bcl-2基因表达。结论:白藜芦醇通过激活PI3K/Akt通路可抑制心肌梗死大鼠细胞线粒体凋亡,并减少I/R引起的心肌细胞损伤。

Objective:To explore the mechanism of Resveratrol in inhibiting mitochondrial apoptosis ofrats with myocardial infarction by activating phosphatidylinositol 3-kinase-protein kinase B(PI3K/Akt)pathway.Methods:Forty-five male SD rats were selected and they were randomly divided into control group,model group and Resveratrol group,15 rats in each group.The ischemia-reperfusion(I/R)model was established in the model group and the Resveratrol group,respectively.Resveratrol was injected in sublingual veinof rats in Resveratrol group for intervention.Detection and comparison of cardiomyocyte injury and apoptosis in rats were implemented.Western blot and real-time quantitative polymerase chain reaction(qRT-PCR)were used to detect the PI3K/Akt pathway and expression levels of mitochondrial apoptotic gene,respectively.Results:The levels of serum troponin T(cTnT)and creatine kinase isoenzyme(CK-MB)of model group were significantly increased,and serum cTnT and CK-MB levels of Resveratrol group were significantly lower than those of model group(t=27.129,t=14.971,P<0.05).The cardiomyocyte apoptosis rate of model group was significantly increased,the cardiomyocyte apoptosis rate ofResveratrol group was significantly lower than that of model group(t=12.687,P<0.05).The PI3K/Akt pathway in cardiomyocyte was significantly inhibited after model was established.Resveratrol could significantly promote the activation of the PI3K/Akt pathway.After model was established,the mRNA level of B lymphocyte tumor-2(Bcl-2)ofcardiomyocytesof rat was down-regulated,whilethe mRNA levels of Bcl-2 interacting mediator of cell death(BIM),Bcl-2-associated X protein(BAX),caspase 9(Caspase9)andCaspase3were up-regulated.Resveratrol could significantly inhibit the gene expressions of BIM,BAX,Caspase9,and Caspase3,and could promote the expression of Bcl-2 gene.Conclusion:Resveratrol can inhibit mitochondrial apoptosis ofrats with myocardial infarction by activating PI3K/Akt pathway and reduce myocardial cell damage caused by I/R.