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目的:研究蟾酥两个主要活性成分对EV71-EGFP病毒感染性克隆的影响,筛选出能抑制EV71-EGFP病毒感染性克隆的天然产物,为治疗由EV71引发的手足口病提供候选预防、治疗药物。方法:分别将含有华蟾酥毒基和酯蟾毒配基的细胞培养液加入到培养RDS细胞的96孔板中,同时加入EV71-EGFP病毒感染性克隆感染。15 h后于倒置显微镜下观察细胞生长状态、荧光显微镜下观察荧光值,并通过多标记微孔板检测系统对EGFP值进行定量研究。结果:与细胞对照组相比,添加蟾酥的两种活性成分虽能减少由EV71-EGFP病毒感染所造成的细胞病变,但仍有部分细胞形态发生变化;能提高细胞的密度和活力;能有效地减少细胞内EV71-EGFP感染性克隆的数量。结论:蟾酥的主要活性成分对EV71-EGFP病毒感染性克隆有一定的抑制作用。
OBJECTIVE: To study the effects of two major active ingredients of toadstool on infectious clones of EV71-EGFP virus and to screen out natural products that inhibit the infectious clones of EV71-EGFP virus and provide candidate prevention and treatment of hand-foot-mouth disease caused by EV71 . METHODS: Cell culture medium containing cinobufagin and resibufogenin was added to 96-well plates of RDS cells, respectively, and infected with EV71-EGFP infectious clone. After 15 h, the growth of cells was observed under an inverted microscope. The fluorescence value was observed under a fluorescence microscope. The EGFP value was quantitatively determined by a multi-labeled microplate detection system. Results: Compared with the cell control group, the two active ingredients added toad could reduce the cytopathic effect caused by infection with EV71-EGFP virus, but some cell morphological changes were still observed; the cell density and vitality were improved; Reduce the number of intracellular EV71-EGFP infectious clones. Conclusion: The main active ingredient of Toadstool can inhibit the infection of EV71-EGFP virus.