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目的:探讨三叶青提取物对人类免疫缺陷病毒(HIV)活性的抑制作用。方法:以不同HIV-1病毒株(实验株HIV-1_(IIIB)、HIV-1耐药株HIV-174V、HIV-1临床分离株HIV-1_(KM018))为研究对象,通过合胞体抑制等多种方法检测三叶青提取物对不同病毒株的抑制作用,MTT法、ELISA法及荧光法分别检测三叶青提取物体外细胞毒性、提取物对HIV-1逆转录酶及蛋白酶活性的抑制作用。结果:三叶青提取物对C8166、MT4、PBMC的体外细胞毒性作用CC50值分别为158.73μg/m L、96.42μg/m L、120.67μg/m L。三叶青提取物对不同HIV-1病毒株有不同程度的抑制作用,EC50值在3.54~78.56μg/m L之间,TI值在2.03~43.18之间;其中实验株HIV-1_(IIIB)急性感染C8166细胞抑制HIV-1 p24抗原表达EC50值最大,C8166细胞形成合胞体TI值最大。提取物对HIV-1慢性感染细胞与正常T淋巴细胞C8166融合阻断、重组HIV-1逆转录酶抑制的EC50值分别为14.79μg/m L、170.15μg/m L。结论:三叶青提取物体外对HIV-1活性有一定的抑制作用,且可能是通过抑制HIV-1病毒至细胞及HIV-1逆转录酶活性达到抗HIV活性的目的。
Objective: To investigate the inhibitory effect of clover leaf extract on human immunodeficiency virus (HIV) activity. Methods: HIV-1 (HIV-1_ (IIIB), HIV-1 HIV-174V and HIV-1 clinical isolates HIV-1_ And other methods to detect the inhibitory effect of clover leaf extract on different virus strains. MTT assay, ELISA and fluorescence were used to detect cytotoxicity of clover leaf extract in vitro, respectively. The effects of extract on HIV-1 reverse transcriptase and protease activity Inhibition. Results: The cytotoxicity of Clover chinense on C8166, MT4 and PBMC were 158.73μg / mL, 96.42μg / mL and 120.67μg / mL respectively. Trichosanthes extract against different HIV-1 strains have different degrees of inhibition, EC50 value between 3.54 ~ 78.56μg / m L, TI value between 2.03 ~ 43.18; experimental strains of HIV-1 (IIIB) The acute infection of C8166 cells inhibited the expression of HIV-1 p24 antigen EC50 maximum, C8166 cells formed syncytia TI maximum. The extracts blocked the fusion of C8166 in chronic infected HIV-1 cells and normal T lymphocytes, and the EC50 values of recombinant HIV-1 reverse transcriptase were 14.79μg / mL and 170.15μg / mL respectively. CONCLUSION: The extract of clover leaf can inhibit the activity of HIV-1 in vitro and may be an anti-HIV activity by inhibiting HIV-1 virus to HIV-1 reverse transcriptase activity.