摘要
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.
出处
《中医药导报》
2011年第3期87-89,共3页
Guiding Journal of Traditional Chinese Medicine and Pharmacy