补骨脂正丁醇部位抗HIV/SIV活性的体外实验研究

In vitro study on anti-HIV/SIV activity of the n-butyl alcohol extract of Psoralea corylifolia Linn

目的观察中药补骨脂乙醇提取物正丁醇部位体外抑制人免疫缺陷病毒(HIV)及猴免疫缺陷病毒(SIV)的活性及作用。方法运用CCK-8试剂检测补骨脂正丁醇部位不同浓度对MT2细胞的细胞毒性。分别以HIV-1BK132和SIVmac251感染MT2细胞为体外实验模型,以细胞病变效应(CPE)、套式反转录荧光定量聚合酶链反应(RT-PCR)法检测细胞内病毒核糖核酸(RNA)相对量和上清中病毒载量为主要评价指标,评估补骨脂正丁醇部位体外抑制HIV-1/SIV的作用及量效关系。结果 CCK-8比色法测定结果显示,补骨脂正丁醇部位最大无毒性浓度为32μg.mL^-1。CPE结果显示,当病毒对照组有大量合胞体(+++)形成时,仅补骨脂正丁醇部位32μg.mL^-1浓度组及16μg.mL^-1浓度组未见合胞体形成(-),与正常细胞对照组和阳性药对照组相当(-),其余各浓度组均有不同程度合胞体形成。RT-PCR结果显示,与病毒对照组相比,补骨脂正丁醇部位在8~32μg.mL^-1浓度范围时,对HIV-1/SIV感染MT2模型中细胞内HIV/SIV RNA的相对量及上清中HIV/SIV病毒载量均有显著的下调作用(P<0.01),且在16~32μg.mL^-1浓度差异尤为显著(P <0.001),与阳性药对照组相当(P> 0.05)。结论补骨脂正丁醇部位对HIV-1 BK132/SIVmac251诱导MT2细胞病变效应有较明显的抑制作用,同时对细胞内及上清液中的HIV/SIV量有显著的下调作用,且在8~32μg.mL-1浓度之间存在一定的量效关系。

Objective To observe the activity and efficiency of the n-butyl alcohol extract of Psoralea corylifolia Linn on inhibiting human immunodeficiency virus(HIV) or simian immunodeficiency virus(SIV) in vitro. Methods CCK-8 reagent was used to detect the cytotoxicity of different concentrations of Psoralen n-butanol extract on MT2 cells. MT2 cells were infected with HIV-1 BK132 and SIVmac251 respectively. To evaluate the inhibitory effect of Psoralen n-butanol extract on HIV-1/SIV in vitro and the dose-effect relationship, the in vitro experimental model was used to detect the cytopathic effect(CPE) and relative amount of viral RNA in the cells and the viral load in the supernatant by RT-PCR. Results The CCK-8 colorimetric assay showed that the maximum non-toxic concentration of Psoralen n-butanol extract was 32μg.mL-1. The results of cytopathic effect(CPE) showed that when the virus control group had a large number of syncytia(+++), no syncytium formation(-) could be found in the 32μg.mL-1 and 16μg.mL^-1 concentration groups of Psoralen n-butanol extract group, equivalent to the normal cell control group and the positive drug control group(-), and the other concentration groups had different degrees of syncytia formation. The results of RT-PCR showed that relative amount of viral RNA in the cells and the viral load in the supernatant were significantly down-regulated in the range of 8-32μg.mL^-1 compared with that of the virus control group(P < 0.01). The difference in concentration of 16-32μg.mL^-1 was particularly significant(P < 0.001), which was equivalent to the positive drug control group(P > 0.05). Conclusion The Psoralen n-butanol extract has a significant inhibitory effect on the pathological effect of HIV-1 BK132/SIVmac251 on MT2 cells. At the same time, there is a significant down-regulation of the amount of HIV/SIV virus in the cells and supernatant, and there is a dose-effect relationship between the concentrations of 8-32μg.mL^-1.