扶芳藤提取物抗HIV-1活性及作用机制  

The anti-HIV-1 activity and mechanisms of Euonymus fortunei extract

作　　者：覃秋珍 蔡丹昭[2] 温奇龙 刘洁[1] 叶力[1] 李雪花 梁浩[1] 刘欣[1] QIN Qiuzhen;CAI Danzhao;WEN Qilong;LIU Jie;YE Li;LI Xuehua;LIANG Hao;LIU Xin(Guangxi Key Laboratory of AIDS Prevention and Control Research,Institute of Sciences,Guangxi Medical University,Nanning,Guangxi 530021,China;Guangxi Key Laboratory of Biomedical Research,School of Basic Medical Sciences,Guangxi Medical University,Nanning,Guangxi 530021,China)

机构地区：[1]广西医科大学生命科学院,广西艾滋病防治研究重点实验室,广西南宁530021 [2]广西医科大学基础医学院,广西高校生物分子医学研究重点实验室,广西南宁530021

出　　处：《中国热带医学》2024年第8期967-971,1005,共6页China Tropical Medicine

基　　金：国家自然科学基金项目(No.31970167);广西自然科学基金项目(No.2024GXNSFBA010134,2023GXNSFBA026264);广西高校中青年教师科研基础能力提升项目(No.2022KY0090)。

摘　　要：目的探究扶芳藤提取物(Euonymus fortunei extract)的抗人类免疫缺陷病毒1(human immunodeficiency virus-1,HIV-1)活性及其作用机制,为研究与开发传统中草药资源治疗获得性免疫缺陷综合征(acquired immunodeficiency syndrome,AIDS)提供理论依据。方法建立外周血单个核细胞(peripheral blood mononuclear cells,PBMCs)-HIV-1_(bal)细胞模型,通过CellTiter-Glo法检测扶芳藤提取物对PBMCs的细胞活性影响,通过HIV-1p24抗原检测测定提取物对HIV-1_(bal)的抑制活性;通过time-of-addition实验及HIV-1关键酶活性抑制实验,探究扶芳藤提取物的抗HIV-1作用机制。结果在PBMCs-HIV-1_(bal)细胞模型中,扶芳藤提取物在最高实验浓度下对细胞活性无明显影响,半数中毒浓度(median cytotoxic concentration,CC_(50))>40μg/mL,在无细胞毒性浓度范围内,随着扶芳藤提取物浓度升高HIV-1 p24水平降低,半数抑制浓度(median inhibitory concentration,IC_(50))=0.084μg/mL,选择指数(selection index,SI)>476.19。Timeof-addition实验发现扶芳藤提取物在与病毒同时加入细胞时对HIV-1IIB活性的抑制率最高,在第2小时对HIV-1活性的抑制率下降,在第18~24小时逐渐失去对病毒的抑制作用,与已知HIV进入抑制剂恩夫韦肽(enfuvirtid,T-20)具有类似的抑制模式。HIV-1关键酶活性实验发现扶芳藤提取物对HIV-1逆转录酶活性无明显的抑制作用,呈现为剂量依赖性抑制HIV-1整合酶活性,随着提取物浓度升高,HIV-1整合酶活性下降(IC_(50)=0.18μg/mL);在一定浓度范围内,提取物可抑制HIV-1蛋白酶活性(IC_(50)=39.18μg/mL)。结论本研究发现扶芳藤提取物具有显著的抗HIV-1活性,可能通过作用于病毒进入细胞和抑制整合酶活性发挥抗HIV-1作用,在抗HIV-1多靶点新药研发中具有较好的应用前景。Objective To investigate the anti-human immunodeficiency virus-1(HIV-1)activity of Euonymus fortunei extract and its mechanism,providing a theoretical basis for the research and development of traditional Chinese herbal resources in the treatment of acquired immunodeficiency syndrome(AIDS).Methods The peripheral blood mononuclear cells(PBMCs)-HIV-1_(bal)cell model system was used as the experimental model,and the effect of Euonymus fortunei extract on the cell activity and anti-HIV-1 activity of PBMCs was investigated by CellTiter-Glo assay and HIV-1 p24 antigen assay.Additionally,the mechanism of anti-HIV-1 action of Euonymus fortunei extract was determined through the time-of-addition assay and HIV-1 enzymatic assays.Results In the PBMCs-HIV-1_(bal)cell model,Euonymus fortunei extract showed no significant effect on cellular activity at the highest concentration,with a median cytotoxic concentration(CC_(50))>40μg/mL.Within the non-cytotoxic concentration range,the level of HIV-1_(bal)decreased with the increase of the extract concentration,with the IC_(50)=0.084μg/mL,and the SI>476.19.Time-of-addition assay revealed that Euonymus fortunei extract had the highest inhibitory activity on HIV-1IIIB when it was added to cells at the same time as the virus,and the inhibitory rate of HIV-1 activity decreased at 2 h and gradually lost its inhibitory effect on the virus at 18-24 h,similar to the known HIV entry inhibitor Enfuvirtid(T-20).Key HIV-1 enzyme activity experiments found that the extract had no significant inhibitory effect on reverse transcriptase activity but exhibited dose-dependent inhibition of integrase activity,with integrase activity decreasing with increasing extract concentration(IC_(50)=0.18μg/mL);The extract could inhibit the activity of HIV-1 protease in a certain concentration range(IC_(50)=39.18μg/mL).Conclusions In this study,it was found that Euonymus fortunei extract has a strong inhibitory activity against HIV-1 activity,which may exert anti-HIV-1 effects by acting on viral entry into ce

关 键 词：扶芳藤提取物 抗人类免疫缺陷病毒1活性 抗人类免疫缺陷病毒1关键酶 

分 类 号：R285.5[医药卫生—中药学]