氧化苦参碱抗流感病毒药效及机制的研究-汕头科学技术协会.DOCVIP

氧化苦参碱抗流感病毒药效及机制的研究 陈小璇，代剑平(，万倩英，朱丹霞，王革非，李康生 汕头大学医学院微生物与免疫学教研室，广东 汕头 515041 目的：研究氧化苦参碱的抗流感病毒活性及其潜在机制。方法：空白对照（DMSO, 0.5%）阳性对照（利巴韦林, 200 μmol/L）氧化苦参碱(12.5、25、50、100 μmol/L)，每组重复6孔。空斑抑制法测定氧化苦参碱抗病毒药效；构建荧光素酶报告质粒，并用Western blotting和ELISA检测氧化苦参碱对流感病毒诱导的细胞TLR4、Myd88和TRAF6启动子转录、TLR4-Myd88- TRAF6- NF-κB信号通路活化及炎性因子释放的影响。结果：氧化苦参碱在12.5－100μmol/L范围内可显著流感病毒复制，其半数有效浓度（EC50）为19.95μmol/L启动子荧光素酶报告质粒检测氧化苦参碱（12.5μmol/L）可显著降低流感病毒诱导的TLR4、MyD88和TRAF6转录比较，。检测氧化苦参碱（12.5μmol/L）可显著降低流感病毒诱导的TLR4-Myd88- TRAF6-NF-κB通路的活化比较，检测氧化苦参碱（12.5μmol/L）可显著降低流感病毒诱导的IL-1β、IL-6、TNF-α、IL-8、IFN-β和IFN-γ释放比较，。结论：氧化苦参碱具抗流感病毒活性，其潜在机制可能与其抑制流感病毒诱导的TLR4-Myd88-TRAF6- NF-κB信号通路活化有关。 氧化苦参碱；流感病毒；TLR4信号通路；炎性因子 Study on the effect and mechanism of oxymatrine on anti- influenza A virus activity CHEN Xiao-xuan, DAI Jian-ping*, WANG Qian-ying, Zhu Dan-xia, WANG Ge-fei, LI Kang-sheng (Department of Microbiology and Immunology, Shantou University Medical College, Shantou 515041, Guangdong, China) [ABSTRACT] OBJECTIVE: This study is to explore the effect and mechanism of oxymatrine on anti-influenza A virus (IAV) activity. METHODS: MDCK and A549 cells were cultured and divided into 4 groups: blank control（DMSO, 0.5%）, virus-treated group, positive group (ribavirin, 200 μmol/L) and oxymatrine-treated group (12.5, 25, 50 and 100 μmol/L) , with 6 repeats of each group. The anti-IAV activity of oxymatrine was determined by plaque inhibition assay. The influence of oxymatrine on the IAV-induced transcription of TLR4, Myd88 and TRAF6, the activation of TLR4-Myd88-TRAF6-NF-κB signal pathway, and the release of inflammatory cytokins were determined by luciferase reporter plasmid, western blotting and ELISA assays, respectively. RESULTS: Plaque inhibition assay showed that oxymatrine, at a range of 12.5～100 μmol/L, could significantly inhibit the replication of IAV in vitro, and its EC50 was 19.95 μmol/L. Luciferase reporter assay showed that oxymatrine, at the concentration of 12.5 μmol/L, could significantly inhibit the transcriptio