目的初步探讨表没食子儿茶素没食子酸酯(EGCG)抗甲型流感病毒的活性及其作用机制。方法采用H5N1假病毒检测体系,观察EGCG对A/Thailand/Kan353/2004H5N1毒株假病毒的抑制作用;采用神经氨酸酶抑制实验进一步分析其作用机制;采用H1N1 FM_1病毒检测体系,根据药物与病毒、细胞的不同作用,通过两种给药方式,评估EGCG对甲型流感病毒FM_1的抑制作用。结果 EGCG能特异性的抑制H5N1假病毒的进入,IC50为145.10μmol.L-1;EGCG对神经氨酸酶具有抑制作用,IC50为417.20μmol.L-1,EGCG对预防和治疗给药方式均有一定的抑制作用,其IC50分别为6.88μmol.L-1和14.83μmol.L-1,治疗指数分别为58.02和26.92。结论 EGCG在体外具有明显的抗甲型流感病毒作用,其作用机制包括抑制病毒的进入和神经氨酸酶活性。
Aim To explore the anti-influenza A virus activity of epigallocatechin gallate (EGCG) and its mechanism. Methods Inhibition of viral-entry by EGCG was determined with luciferase assay in MDCK cells infected with A/Thailand/Kan353/2004 H5N1 pseudovirus. Nneuraminidase inhibition assay was further used to investigate the mechanism of anti-influenza activity. Anti-influenza activity of EGCG was further confirmed in MDCK cells infected with influenza A virus FM_1 strain with different medications, including treated, prophylactic medications. Results EGCG inhibited the entry of H5N1 pseudovirus into MDCK cells with IC50 of 145.10 μmol · L^-1. EGCG also inhibited the activity of neuraminidase with IC50 of 417.20 μmol · L^-1. EGCG also showed anti-influenza viral effect on FM_1 virus in MDCK cells. ICso was 6. 88 μmol · L^-1 when added before inoculation with influenza A virus FM_ 1 strain, while IC50 was 14. 83 μmol · L^-1 when added after the virus inoculation. TI was 58.02 and 26.92, respectively. Conclusions EGCG can inhibit influenza A virus in vitro. The mechanism of EGCG includes at least the virus entry and neuraminidase activity inhibition.