金叶败毒颗粒抑制人巨细胞病毒vMIA蛋白表达及作用的实验研究

Experimental study on the inhibiting effect of Jinyebaidu granules on the expression of human cytomegalovirus vMIA protein

目的:通过研究金叶败毒颗粒对人巨细胞病毒vMIA蛋白表达及作用的调节,探讨金叶败毒颗粒体外对HCMV感染的预防、阻断和治疗效应及其机制。方法:采用细胞形态观察测定病毒的感染性。应用免疫荧光检测vMIA蛋白在病毒感染过程中表达的规律,在病毒感染不同阶段(病毒吸附前、吸附时和吸附后)应用金叶败毒颗粒,观察药物对HCMV vMIA蛋白表达水平的影响。转染pcDNA UL37xl质粒,经G418筛选,Western blot鉴定vMIA的表达。应用流式细胞术检测药物对TNFα+CHX攻击后细胞凋亡指数的影响。结果:HCMV感染后4 h vMIA开始表达,在病毒感染48h表达最显著。金叶败毒颗粒可显著抑制HCMV vMIA蛋白的表达,其作用较更昔洛韦弱。金叶败毒颗粒可以恢复被vMIA抑制的TNFα诱导的凋亡,但更昔洛韦无此功效。结论:金叶败毒颗粒可显著抑制HCMV vMIA蛋白的表达及其作用,这可能是金叶败毒颗粒抗HCMV效应的主要作用机制之一。

Objective：To explore the preventive,inhibiting and therapeutic effects of Jinyebaidu granules on human cytomegalovirus（HCMV） infection and the mechanism by researching the regulatory effect of Jinyebaidu granules on expression of HCMV vMIA protein.Methods：The infectivity of HCMV were observed by cytological morphologic method.Immunofluorescence was used to detect the expression regularity of vMIA protein in the course of HCMV infection,Jinyebaidu granules were used at different periods of HCMV infection（before,during,and after adsorption of HCMV）,the effect of Jinyebaidu granules on expression level of HCMV vMIA protein was observed.After transfected by pcDNA UL37xl plasmid and G418 screening,Western blot was used to identify the expression of vMIA protein.Flow cytometry was used to detect the effect of Jinyebaidu granules on cell apoptosis index after attacked by TNF-α＋CHX.Results：At 4 hours after HCMV infection,vMIA expression begun,then peaked at 48 hours after HCMV infection.Jinyebaidu granules inhibited the expression of HCMV vMIA protein significantly,and its effect was weaker than that of ganciclovir.Jinyebaidu granules could restore TNF-α-induced apoptosis inhibited by vMIA,but ganciclovir didn＇t have the effect.Conclusion：Jinyebaidu granules can inhibit the expression of HCMV vMIA protein significantly,which may be one of the main mechanisms of anti-HCMV effect of Jinyebaidu granules.