益气活血中药复方对CVB_3病毒性心肌炎心肌细胞损伤的影响

Effect of Yiqihuoxue Chinese Herbal Compound on Myocardial Injury of CVB_3 Viral Myocaralitis

目的观察益气活血中药复方对CVB3病毒性心肌炎心肌细胞损伤的影响。方法以体外细胞培养方法,建立CVB3病毒性心肌炎心肌细胞感染模型。将细胞分为正常组、病毒组、中药组。采用荧光探针Hoechst33258染色法检测心肌细胞凋亡;采用激光共聚焦显微镜检测心肌细胞内钙离子浓度;采用SABC免疫组化法检测心肌细胞心肌肌钙蛋白(cTnI)表达;采用透射电镜检测各组心肌细胞超微结构。结果病毒组与正常组心肌细胞相比,细胞超微结构损伤严重,并可见明显的凋亡细胞核形态,而中药组心肌细胞形态及细胞器结构接近正常组,未见明显的凋亡细胞核形态;正常组心肌细胞cTnI呈阴性表达,病毒组cTnI呈强阳性表达,与正常组比较,差异有统计学意义(P<0.05),中药组cTnI呈弱阳性表达,与病毒组比较,差异有统计学意义(P<0.01);正常组心肌细胞经中药干预后,钙离子荧光强度无明显变化(P>0.05),而病毒组心肌细胞经中药干预后,钙离子平均荧光强度升高(P<0.01),与正常组比较,差异无统计学意义(P>0.05)。结论益气活血中药复方可抑制CVB3感染后细胞凋亡发生,抑制cTnI蛋白表达,防御并减轻CVB3造成的心肌损伤,具有修复损伤心肌细胞超微结构、改善心肌细胞舒缩功能和血液动力学作用。

Objective To observe the impact of Yiqihuoxue Chinese herbal compound on myocardial injury of CVB3 viral myocarditis.Methods In vitro cell culture,myocardial CVB3 viral infection model was established.Cells were divided into normal group,virus group and Chinese herb intervention group.Fluorescent probe staining Hoechst33258 was used to detect cardiomyocyte apoptosis.Confocal laser microscopy was used to determine myocardial intracellular calcium ion concentration.By SABC immunohistochemical method,expression of cTnI of myocardial cells was detected.Myocardial ultrastructure was observed by using transmission electron microscopy.Results Compared with normal group,cell ultrastructure injury of virus group was serious and showed obvious nuclear morphology of apoptosis.Myocardial morphology and organelle structure of Chinese herb intervention group was close to the normal group,no obvious nuclear morphology of apoptosis.cTnI expression of myocardial cells of normal group was negative.The virus group showed a strong expression of cTnI,the difference was significantly compared with the normal group（P〈0.05）.Chinese herb intervention group showed a weak positive cTnI expression,the difference was significantly compared with the virus group（P〈0.01）.Normal myocardial cells intervented by Chinese herb had no significant calcium fluorescence intensity change（P〉0.05）.After the Chinese herb intervention,mean fluorescence intensity of calcium increased in myocardial cells of the virus group（P〈0.01）,with no significant difference compared with the normal group（P〉0.05）.Conclusion Yiqihuoxue Chinese herbal compound can inhibit apoptosis,inhibit the expression of cTnI,defense and to reduce myocardial injury of CVB3 infected cells of caused by CVB3.It can repair the damage of myocardial ultrastructure,protect the myocardium and improve myocardial contractile function and hemodynamics.