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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: To study the in vitro anti-simian immunodeficiency virus (SIV) activity of Ganoderma lucidum spore water extract (AEGLS) and its mechanism of action. Methods: The number of syncytia was counted after adding TCID50 SIV virus dilution and AEGLS dilution to CEM × 174 cells (CEM T lymphocytes and 174 B lymphocyte hybrid cells) for 4 days. The numbers of syncytium in AEGLS anti-SIV active. Using different dosing time experiment, the role of stage analysis experiments, Western blot analysis of its mechanism of action. Results: AEGLS had good anti-SIV activity with IC50 of (466.62 ± 20.21) mg · L-1 and lower cytotoxicity. The half-cell cytotoxicity concentration (CC50) was 3.13 ± 0.11 g · L-1 , The selection index of 6.71. Anti-SIV mechanism studies show that AEGLS plays a major role in the early stages of SIV virus infection, and inhibition of SIV virus adsorption or cell penetration, Western blot showed that AEGLS can reduce the SIV capsid protein P27 (SIV P27) expression levels. CONCLUSION: AEGLS has a good anti-SIV effect and its main role in the early stages of SIV virus infection is related to inhibition of SIV virus adsorption or cell penetration, and can reduce the expression level of SIV P27 capsid protein.