达塞布韦抗黄热病毒感染和涉及初步机制的研究  

Antiviral activity of dasabuvir in yellow fever virus infection:A preliminary mechanistic study

作　　者：钱汐晶 刘燕 吴兵安 戚中田 赵平 Qian Xijing;Liu Yan;Wu Bing’an;Qi Zhongtian;Zhao Ping(Division of Biomedical Protection,Department of Naval Medicine,Naval Medical University,Shanghai 200433,China)

机构地区：[1]海军军医大学海军医学系生物医学防护教研室,上海200433

出　　处：《广州医科大学学报》2022年第2期18-23,共6页Academic Journal of Guangzhou Medical University

摘　　要：目的:检测达塞布韦抗黄热病毒(YFV)感染的效果并对其涉及的机制进行初步探索。方法:用不同浓度的达塞布韦处理人肝癌Huh7细胞,检测达塞布韦对细胞的毒性并计算细胞半数毒性浓度(CC50)。在安全浓度范围下,检测不同浓度的达塞布韦对YFV感染的作用并计算半数有效抑制浓度(IC50)。在YFV感染的不同时间段加入达塞布韦,检测其抗YFV的起效阶段。利用带有报告基因的YFV复制子,评价达塞布韦对YFV复制的影响。通过表达YFV NS5蛋白,利用BIAcore亲和力分析检测小分子达塞布韦与病毒NS5蛋白之间的相互作用。结果:达塞布韦细胞毒性低(CC50:346.1μM),能有效抑制YFV感染(IC50:7.253μM,P<0.01)。动力时间窗实验表明,达塞布韦主要在感染后期起效,能有效干扰YFV复制子在靶细胞内的复制效率(P<0.01)。BIAcore检测进一步证实了达塞布韦通过与YFV NS5蛋白相互作用抑制了病毒的复制。结论:达塞布韦可阻断YFV感染靶细胞,作用机制为通过与YFV NS5相互作用,干扰复制关键酶的活性,抑制病毒复制。Objective:To investigate the effect of dacebuvir against yellow fever virus(YFV)infection and to preliminarily explore the mechanism involved.Methods:Human hepatoma Huh7 cells were treated with different concentrations of dasabuvir,and thereby,the cytotoxicity of dasabuvir was measured,and the half maximal cytotoxic concentration(CC50)was calculated.Within the safety concentration range,the effect of different concentrations of dasabuvir on YFV infection was examined and the half maximal inhibitory concentration(IC50)was calculated.Dasebuvir was added and incubated during different periods of YFV infection to detect the time window of its anti⁃YFV action.Using a YFV replicon with a reporter gene,the effect of dasabuvir on YFV replication was evaluated.The YFV NS5 protein was expressed and BIAcore affinity analysis was used to detect the interaction between small molecules of dasabuvir and viral proteins.Results:Dasebuvir showed low cytotoxicity(CC50:346.1μM)and could effectively inhibit YFV infection(IC50:7.253μM,P<0.01).The effect analysis time window showed that the onset of dasabuvir activity largely occurred during the late stage of infection,and interfered effectively with the replication efficiency of YFV replicon in target cells(P<0.01).BIAcore assay further confirmed that dasabuvir inhibited viral replication by interacting with YFV NS5 protein.Conclusion:Dasebuvir can inhibit the YFV infection of target cells via interaction with NS5,thereby interfering with the activities of critical enzymes that relate to the virus replication.

关 键 词：黄热病毒 聚合酶抑制剂 抗病毒药物 达塞布韦 

分 类 号：R373[医药卫生—病原生物学]