灵芝体外抑制猴免疫缺陷病毒作用的研究

Study on Effects of Inhibition of Ganoderma lucidum on Simian Immunodeficiency Virus in vitro

目的:对灵芝孢子水提物(AEGLS)进行体外抗猴免疫缺陷病毒(SIV)活性分析并对其作用机制进行初步研究。方法:300 TCID50的SIV病毒稀释液和AEGLS稀释液同时加入到CEM×174细胞(CEM T淋巴细胞和174 B淋巴细胞杂交细胞)中培养4 d后计数合胞体数量并进行统计,研究AEGLS抗SIV活性。利用不同加药时间点实验、作用阶段分析实验、Western blot对其作用机制进行初步分析。结果:AEGLS具有良好的抗SIV活性,其半数抑制浓度IC50为(466.62±20.21)mg·L-1,并且其细胞毒性较低,半数细胞毒性浓度CC50为(3.13±0.11)g·L-1,选择指数为6.71。抗SIV机制研究显示AEGLS主要作用在SIV病毒感染的早期阶段,与抑制SIV病毒的吸附或者穿透细胞有关,Western blot实验显示AEGLS能降低SIV衣壳蛋白P27(SIV P27)的表达水平。结论:AEGLS具有良好的抗SIV作用,其主要作用在SIV病毒感染的早期阶段,与抑制SIV病毒的吸附或者穿透细胞有关,并且能降低SIV P27衣壳蛋白的表达水平。

Objective： This paper presents the effects of anti simian immunodeficiency virus （SIV） of the aqueous extract from Ganodorma lucidum spores （AEGLS） by using SIVmac and CEM ×174 cells as a model in vitro, and the mechanism were preliminarily studied. Method： The inhibitory effects of AEGLS on SIV were observed when SIV and CEM × 174 system were employed as the models, and the mechanism of action were preliminarily studied by using time course assay, effective stage analysis, Western blot. Result： AEGLS had a remarkable anti-SIV activity. The IC50 （50% inhibitory concentration） of AEGLS was （466.62 ± 20.21 ） mg. L^-1, and it was low cytotoxicity which its CC50 （50% cellular cytotoxicity） was （3.13 ±0. 11） g. L^-1 and selection index was 6.71. The results of anti-SIV mechanism showed AEGLS main acted on the early stage of SIV infection, and might contribute to affect the SIV attachment and/or penetration into the ceils. The results ofWestern blot showed AEGLS could decrease SIV P27 remarkable anti-SIV activity and is main acted on the the SIV attachment and/or penetration into the cells, capsid protein expression level. Conclusion： AEGLS had a early stage of SIV infection, and might contribute to affect and could decrease SIV P27 capsid protein expression level.