广枣总黄酮对CVB_3病毒引起细胞凋亡的影响

The effect of TFC to cell apoptosis induced by CVB3 virus ABSTRACT

目的:建立病毒性心肌炎的体外模型,用新的化学方法提取到含量较高的广枣总黄酮,研究广枣总黄酮抗病毒的效果。方法:化学法提取广枣总黄酮,高效液相法测定其含量。培养HeLa细胞和乳鼠心肌细胞,在HeLa细胞和心肌细胞水平上对柯萨奇B3病毒进行毒力滴定,并测定广枣总黄酮的药物毒性,建立体外病毒性心肌炎模型,通过细胞病变抑制实验测定广枣总黄酮体外抗柯萨奇病毒能力。采用Hoechst33258荧光单染观察染色质凝聚,观察病毒性心肌炎发病过程中心肌细胞是否存在凋亡,以及运用流式细胞仪检测广枣总黄酮是否抑制细胞凋亡。统计学方法采用T检验。结果:成功建立体外病毒性心肌炎的模型,广枣总黄酮在HeLa细胞和乳鼠心肌细胞实验中可有效抑制CVB3病毒引起的细胞病变数(P<0.05),对病毒繁殖及细胞凋亡有抑制作用。结论:成功建立体外细胞模型,通过体外实验证实提取的广枣总黄酮对心肌细胞有保护作用,通过凋亡相关实验证实病毒性心肌炎发病过程中有心肌细胞凋亡,且广枣总黄酮能抑制心肌细胞凋亡作用。

[Objectives]To establish model of viral myocarditis in vitro,To distill TFC（Total flavones of choerospondias axillaries fructus） with a new method,To test the anti-viral effect of TFC.Method：Using chemical means to distill TFC and testcontent with HPLC method.TCID50 of CVB3（coxsackievirus B3） was determined with Reed-Muench method,the anti-viral effect of TFC in vitro was validated through cytopathic effect experiment.Cardiomyocyte Apoptosis was performed by using Hoechst 33258、Flow cytometry.Result：The viral myocarditis model in vitro was successfully established,TFC can inhibit CPE（P0.05）in CPE experiment.Virus-reproducing inhibiting experiment shows TFC can effectively inhibit coxsackievirus,Logarithm Value raise more than one unit,Apoptosis rate of cells was reduced.Conclusion：we established viral mycarditis model in vitro and proved TFC is an effective antiviral drug,TFC can reduce the rate of apoptosis induced by CVB3.