论文部分内容阅读
目的了解中草药或制剂槲皮素、甘草酸二铵和青蒿琥酯对艾滋病病毒复制的影响。方法在C8166T细胞被携带分泌型荧光素(Gaussia luciferase,G-Luc)的HIV病毒感染的同时或2h后加入3种不同浓度的中草药(槲皮素、甘草酸二铵和青蒿琥酯),2~3d后观察细胞形态变化及由病毒感染导致的膜融合所至合胞体的形成,并采用荧光素酶报告基因检测C8166T细胞培养上清中的病毒相关G-Luc活性。结果 C8166T细胞的G-Luc结果显示,与甘草酸二铵、槲皮素组相比,而青蒿琥酯组的荧光活性酶活性增幅最大(F=96.126,P<0.01)。形态学观察显示:经青蒿琥酯处理的由病毒感染导致的膜融合所形成的合胞体,与对照组相比数目明显增多,且体积更大。Western blot检测表明C8166T细胞内的HIV-1核衣壳蛋白P24水平增强;ELISA检测显示该细胞培养上清中的HIV-1P24的含量与对照组相比有明显升高(F=9.503,P<0.05),提示青蒿琥酯处理可增强病毒的表达或释放。在用青蒿琥酯处理静止期人外周血单核细胞(PBMCs)后,ELISA检测细胞培养上清中的P24含量显示,0.078μg/ml青蒿琥酯组的P24含量较对照组明显增加(t=-14.354,P<0.01),HIV-1潜伏感染明显增加。Real-time PCR结果表明,在C8166T细胞系中,0.078μg/ml青蒿琥酯组的病毒RNA明显多于对照组(t=115.086,P<0.01),证实青蒿琥酯能提高细胞内HIV-1基因转录水平。结论青蒿琥酯可提高C8166T细胞和人PBMC中HIV-1基因的转录,具有潜在的促进HIV感染的作用。
Objective To understand the effects of Chinese herbal medicine or quercetin, diammonium glycyrrhizinate and artesunate on HIV replication. Methods Three different concentrations of Chinese herbal medicine (quercetin, diammonium glycyrrhizinate and artesunate) were added simultaneously with or after 2 hours after infection of HIV-1 infected cells with C8166T cells by G-Luc. After 2 ~ 3d, morphological changes of cells and the formation of syncytium by membrane fusion were observed. The luciferase reporter gene was used to detect the activity of virus-related G-Luc in the culture supernatant of C8166T cells. Results The G-Luc results of C8166T cells showed that compared with diammonium glycyrrhizinate and quercetin group, the activity of fluorescent enzyme in artesunate group increased most (F = 96.126, P <0.01). Morphological observations showed that the syncytia formed by membrane fusion by virus-infected artemisinin treatment significantly increased in number and larger in volume than the control group. Western blot showed that the level of HIV-1 nucleocapsid protein P24 in C8166T cells was enhanced. ELISA assay showed that the content of HIV-1 P24 in the culture supernatant was significantly higher than that in the control group (F = 9.503, P < 0.05), suggesting that artesunate treatment can enhance the expression or release of the virus. After treatment of stationary human peripheral blood mononuclear cells (PBMCs) with artesunate, the detection of P24 in the cell culture supernatant by ELISA showed that the P24 content in the 0.078 μg / ml artesunate group was significantly increased compared to the control group t = -14.354, P <0.01), HIV-1 latent infection increased significantly. Real-time PCR results showed that in the C8166T cell line, the viral RNA of 0.078μg / ml artesunate group was significantly more than that of the control group (t = 115.086, P <0.01), confirming that artesunate can increase intracellular HIV -1 gene transcription level. Conclusion Artesunate can enhance the transcription of HIV-1 gene in C8166 T cells and human PBMCs, which has the potential to promote HIV infection.