蒙药漏芦花不同提取物体外抗甲型H1N1流感病毒活性研究

Study on the anti-influenza A(H1N1) virus activity of different extracts from Mongolian Rhaponticum uniflorum(L.) DC. in vitro

目的:研究蒙药漏芦花不同提取物体外抗甲型H1N1流感病毒的作用。方法:制备蒙药漏芦花不同浓度乙醇提取物,乙酸乙酯提取物和水煎液,利用SPR-Biacore技术筛选与H1N1病毒膜蛋白(HA)、核蛋白(NP)高耦联的漏芦花提取物,CCK8法测定其最大无毒剂量;给药奥司他韦及蒙药漏芦花不同提取物,ELISA法测定病毒抑制率,Probit法回归半数有效剂量(IC50)及治疗指数(TI);MacSynergyⅡ软件分析联合作用效果。结果:漏芦花乙酸乙酯在1.25mg/mL时抗病毒效果最佳,病毒抑制率为64.59%(P<0.01),联合用药实验表明奥司他韦与漏芦花乙酸乙酯提取物具有高度协同作用。结论:漏芦花的乙酸乙酯提取物具有较强体外抑制甲型流感H1N1病毒作用,与奥司他韦联合用药具有高度协同作用,对临床抗病毒药物配伍具有指导意义。

Objective: To study the effects of different extracts from Mongolian Rhaponticum uniflorum(L.) DC.(LLH) on anti-influenza A(H1 N1) virus in vitro. Methods: The different ethanol extraction, ethyl acetate extraction and aqueous extraction of LLH were prepared for test;SPR-Biacore technique was used to screen the different extractions of LLH, the maximum non-toxic dose was evaluated by CCK8 test. Based on the result of CCK8 test, MDCK cells were infected with H1N1 virus and administered with oseltamivir and different extractions of LLH, the viral inhibition rate was measured by influenza a virus antigen ELISA kit, the half of effective dose(IC50) and treatment index(TI) were calculated by the Probit method.Synergistic effect was analyzed by MacSynergy Ⅱ software. Results: Ethyl acetate extraction of LLH had the best antiviral effect at 1.25 mg/mL, with a virus inhibition rate of 64.25%(P<0.01). There is a high synergistic effect on the combination of Oseltamivir and Ethyl acetate extraction of LLH. Conclusion: The ethyl acetate extraction of LLH has a strong inhibitory effect on anti-influenza A(H1N1) virus in vitro, and has a highly synergistic effect with oseltamivir, which has a guiding significance for the compatibility of clinical antiviral drugs.