OBJECTIVE To observe the effect mechanism of Qingtouxiere Decoction on anti-influenza A H1N1 influenza virus in vitro.METHODS The macrophages were stimulated with FM1 in the lungs of influenza A virus, respectively, with different concentrations of 0.388mg/mL, 0.194mg/mL, 0.097mg/mL, 0.0485mg/mL and 0.02425mg/mL.The expression of TNF-α, IP-10 and IL-6, TLR-7, mRNA and protein expression of MyD88 and NF-κB.RESULTS The expression of TNF-α, IP-10 and IL-6 in macrophages of Ana-1 mice was inhibited by Qingtouxiere Decoction, and the infection of H1N1 virus was decreased in 0.02425mg/mL group (P<0.01).The level of TNF-α was significantly lower in the group of Qingtouxiere Decoction 0.097mg/mL (P <0.01).The expression of TLR-7, MyD88 and NF-κB mRNA and protein were increased after Ana-1 macrophage infection in H1N1 influenza virus group.The expression of TLR-7, MyD88 and NF-κB mRNA was increased by 0.0485mg/mL group (P<0.01).The expression of MyD88 mRNA in the H1N1 infected group was significantly higher than that in the control group (P <0.01).The expression of TLR-7 mRNA was significantly inhibited The expression of NF-κB mRNA in H1N1 virus group was significantly higher than that in H1N1 virus group (P<0.01).The expression of TLR-7 protein in H1N1 virus group was significantly higher than that in H1N1 virus group(P<0.01).The expression of NF-κB in the H1N1 infected group was significantly higher than that in the H1N1 virus group (P<0.01).The expression of NF-κB in the H1N1 infected group was significantly higher than that in the control group (P<0.01).CONCLUSION The activation of TLR-7/MyD88/NF-κB signaling pathway induced by H1N1 influenza virus can inhibit the expression of downstream inflammatory factors and play the antiviral effect.%目的 研究清透邪热方体外抗甲型H1N1流感病毒效应机制.方法 将清透邪热方制备成0.388、0.194、0.097、0.0485、0.02425mg/mL不同浓度,用H1N1甲型流感病毒感染小鼠Ana-1巨噬细胞,给予不同浓度清透邪热方及达菲进行干预,检测肿瘤坏死因子α(TNF-α)、干扰素诱导蛋白-10(IP-10)和白细胞介素-6(IL-6)表达,Toll样受体7(TLR7)及其下游髓样分化因子88(MyD88)、核因子-κB(NF-κB)mRNA和蛋白表达.结果 清透邪热方能够抑制甲型H1N1流感病毒感染Ana-1细胞TNF-α、IP-10、IL-6的表达;其中清透邪热方0.02425mg/mL组降低H1N1病毒感染组TNF-α、IL-6显著(P<0.01);清透邪热方0.097mg/mL组降低IP-10显著(P<0.01).清透邪热方能够抑制H1N1流感病毒感染后Ana-1细胞后TLR-7、MyD88及NF-κB mRNA及蛋白表达升高;其中清透邪热方0.0485mg/mL组抑制H1N1病毒感染组TLR-7 mRNA、MyD88 mRNA表达显著(P<0.01);清透邪热方0.02425mg/mL组抑制H1N1病毒感染组NF-κB mRNA表达显著(P<0.01);清透邪热方0.0485mg/mL组抑制H1N1病毒感染组TLR-7蛋白表达升高显著(P<0.01),清透邪热方0.388mg/mL组抑制H1N1病毒感染组MyD88蛋白、NF-κB蛋白表达显著(P<0.01).结论 清透邪热方通过抑制甲型H1N1流感病毒感染Ana-1细胞TLR-7、MyD88及NF-κB mRNA及蛋白表达升高,减少TNF-α、IP-10和IL-6表达,起到抗病毒作用.
展开▼
