目的 研究蒲葵籽提取物的体外抗HIV-1活性,对有活性粗提物进行初步机制研究。方法 采用细胞毒性、合胞体抑制、HIV-1感染细胞保护实验和HIV-1 p24抗原测定等实验对蒲葵籽提取物体外抗HIV-1活性进行筛选和确认;采用重组HIV-1逆转录酶和蛋白酶活性抑制实验,融合阻断实验初步探讨活性粗提物的作用机制。结果 蒲葵籽的醋酸乙酯(P3)提取物具有较强的体外抗HIV-1活性,P3抑制HIV-1诱导C8166细胞形成合胞体的EC50为5.64μg/mL,对C8166细胞的毒性较小,CC50大于200μg/mL,治疗指数(TI)大于35.46;P3抑制HIV-1急性感染中p24抗原表达的EC50为23.04μg/mL,抑制正常C8166细胞与慢性感染细胞H9/HIV-1 B融合的EC50为8.00μg/mL;P3在质量浓度为200μg/mL时,对HIV-1逆转录酶的抑制率为28.86%;P3抑制重组HIV-1蛋白酶活性的EC50为1.77μg/mL。结论 蒲葵籽的醋酸乙酯提取物(P3)具有较强的体外抗HIV-1活性,其作用机制可能主要为阻断病毒进入和抑制HIV-1蛋白酶活性。

Objective ?To evaluate in vitro anti-HIV-1 activities of the extracts from Livistona chinensis?seeds.Methods ?To screen and confirm the anti-HIV-1 activities of the?exracts from L.chinensis seeds.?thecytotoxicity assay,syncytium reduction assay,protection for HIV-1 induced lytic assay,and ELISA?for HIV-1 p24 antigen?assay were used.The?inhibition assay of recombinant HIV-1 RTactivityand?protease activity,and co-cultivation assaywere used tostudy the primarymechanisms.Results ?Theethyl?acetate extract(P3)from L.chinensis seeds inhibited HIV-1 potentlywith atherapeutic index(TI)over?35.46.The P3 showed the anti-HIV-1activitywith EC50 of5.64/19/mLinsyncytiaformationassayand?exerted cytotoxicity againstC8166 cells withCC50>200弘g/mL.Inacute infection,the P3 inhibited HIV-?1p24 antigen expressionwithEC50of23.04弘g/mL.The P3 also inhibited cell-to-cell H9/HIV-1IBfusion?in CO-culture with ECso of 8.00pg/mL.TheP3 at?a concentration of 200 pg/mL weaklyinhibited the?recombinant HIV-l RT activity by 28.86%.Italso inhibitedsignificantlythe recombinant HIV-1 protease?activitywith?EC50 of?1.77 pg/mL.Conclusion ?The P3?is?effectiveagainstHIV-1.The anti-HIV-1?mechanism of P3mightrelate to the blockade of HIV-1 entry into cells in order?to inhibit the activity of?HIV-1 protease.?

国家科技攻关计划资助项目(2004BA719A14);中国科学院知识创新工程重要方向资助项目(KSCX1-YW-R-15KSCX1-YW-R-24);云南省科技攻关计划资助课题(2004NG12)