黄芪总黄酮对柯萨奇B3病毒感染心肌细胞内质网应激及促凋亡信号因子作用

Effect of total flavonoids of astragalus on endoplasmic reticulum chaperone,apoptosis signaling factor in mouse myocardium with myocarditis caused by coxsackievirusB3

目的:研究黄芪总黄酮对柯萨奇B3病毒感染心肌细胞内质网应激伴侣蛋白GRP78、GRP94及促凋亡信号分子JNK、p-JNK、Caspase 12表达的影响。方法:实验分3组,对照组(正常细胞)、柯萨奇病毒感染组(正常细胞+柯萨奇B3病毒)、黄芪总黄酮组(正常细胞+柯萨奇B3病毒+黄芪总黄酮)。采用免疫组织化学方法检测培养乳鼠心肌细胞α-SMA蛋白,采用Western blotting技术检测各组心肌细胞内质网应激伴侣蛋白GRP78、GRP94、p-JNK、JNK及Caspase 12表达。结果:1与对照组比较,柯萨奇B3病毒感染组心肌细胞GRP78、GRP94表达增多(P<0.01),黄芪总黄酮组心肌细胞GRP78、GRP94表达减少(P<0.01)。与对照组比较,柯萨奇B3病毒感染组心肌细胞p-JNK及Caspase 12表达增多(P<0.01),黄芪总黄酮组心肌细胞p-JNK及Caspase 12表达减少(P<0.01)。各组心肌细胞JNK表达无差异(P>0.05)。结论:柯萨奇B3病毒可引发心肌细胞内质网应激,并使促凋亡信号分子p-JNK、Caspase 12表达增多,引发心肌细胞凋亡;黄芪总黄酮抑制柯萨奇B3病毒感染心肌细胞内质网应激,并抑制心肌细胞凋亡,该实验结果可能是其对病毒性心肌炎的作用机制之一。

Objective：To investigate the effect of total flavonoids of astragalus on endoplasmic reticulum chaperone,apoptosis signaling factor.Method：The suckling mouse myocardium of primary culture were randomly divided into control group,coxsackievirusB3 infecton group and total flavonoids of astragalus group.Firstly,we identifyed the suckling mouse myocardium.α-SMA was monitored by immunohistochemical method.Then the expression change of endoplasmic reticulum chaperone GRP78,GRP94,p-JNK,JNK and Caspase-12 was detected by Western blotting.Result：1Compared with that of control group,the GRP78,GRP94,p-JNK and Caspase-12 expression in coxsackievirusB3 infecton group was improved（P〈0.01）.2Compared with coxsackievirusB3 infecton group,GRP78,GRP94,p-JNK and Caspase-12 expression was decreased in total flavonoids of astragalus group（P〈0.01）.JNK of each group was not different in expression（P〉0.05）.Conclusion：CoxsackievirusB3infection can cause endoplasmic reticulum stress of myocardium and induced mouse myocardium apoptosis.But total flavonoids of astragalus can control the effect of CoxsackievirusB3,inhibiting myocardium apoptosis and this may be the mechanism of total flavonoids of astragalus effects coxsackievirusB3.