连翘苷对H1N1病毒株感染小鼠的作用

Effect of Forsythin on H1N1 Virus Strain-Infected Mice

目的研究连翘苷对H1N1病毒株感染小鼠的影响。方法60只H1N1病毒株感染小鼠经随机数字法分为感染组、连翘苷组、连翘苷+罗唑利宾(Loxoribine)[Toll样受体7(TLR7)/髓样分化因子88(MyD88)信号通路激活剂]组,各20只;20只健康小鼠滴鼻等体积0.9%氯化钠溶液,设为正常组。流式细胞仪检测外周血CD4+、CD8+T淋巴细胞占比;实时荧光定量聚合酶链反应(qRT-PCR)法检测肺组织病毒相对表达量;Western blot法检测肺组织TLR7、MyD88蛋白表达量。结果与感染组比较,连翘苷组外周血CD8+T淋巴细胞占比、肺组织病毒相对表达量、TLR7、MyD88蛋白表达量降低,外周血CD4+T淋巴细胞占比、CD4+/CD8+升高(P<0.05);与连翘苷组比较,连翘苷+Loxoribine组外周血CD8+T淋巴细胞占比、肺组织病毒相对表达量、TLR7、MyD88蛋白表达量升高,外周血CD4+T淋巴细胞占比、CD4+/CD8+降低(P<0.05)。结论连翘苷可抑制H1N1病毒株感染小鼠病毒复制,改善肺功能及免疫功能,可能通过抑制TLR7/MyD88信号通路活性降低炎性细胞因子水平,从而保护H1N1感染小鼠。

Objective To study the effect of phillyrinon immune function indexes and inflammatory factors in mice infected with H1N1 virus strain.Methods Sixty H1N1 virus-infected mice were randomly divided into infection group,phillyrin group,and phillyrin+Loxoribine(TLR7/MyD88 signaling pathway activator)group,20 mice in each group;another 20 healthy mice were selected as the normal group by intranasal instillation of the same volume of normal saline.Flow cytometry was used to detect the proportion of CD4+and CD8+T lymphocytes in peripheral blood.The relative expression of virus in lung tissue was detected by real-time quantitative polymerase chain reaction(qRT-PCR).Western blot was used to detect the expression of TLR7 and MyD88 in lung tissue.Results Compared with the infection group,the proportion of CD8+T lymphocytes in peripheral blood,the relative expression of virus in the lung tissue,and TLR7,MyD88 protein expression were decreased,the proportion of peripheral blood CD4+T lymphocytes,CD4+/CD8+were increased in the phillyrin group(P<0.05).Compared with the phillyrin group,the proportion of CD8+T lymphocytes in peripheral blood,the relative expression of virus in the lung tissue,and TLR7,MyD88 protein expression were increased,the proportion of peripheral blood CD4+T lymphocytes,CD4+/CD8+were decreased in the phillyrin+Loxoribine group(P<0.05).Conclusion Phillyrin can inhibit H1N1 virus replication,improve lung function and immune function,and reduce the level of inflammatory factors,thereby protecting H1N1-infected mice by inhibiting the activity of TLR7/MyD88 signaling pathway.