清透邪热方对甲型H1N1流感病毒感染Ana-1细胞TLR-7、MyD88及NF-κB mRNA及蛋白表达的影响

Effects of Qingtouxiere Decoction on Expression of TLR-7,MyD88 and NF-κB mRNA and Protein in Ana-1 Cells of Influenza A H1N1 Influenza Virus

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、0.024 25mg/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0.024 25mg/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0.048 5mg/mL组抑制H1N1病毒感染组TLR-7mRNA、MyD88mRNA表达显著(P<0.01);清透邪热方0.024 25mg/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0.048 5mg/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。

OBJECTIVE To observe the effect mechanism of Qingtouxiere Decoction on anti-influenza A HIN1 influenza vi- rus in vitro. METHODS The macrophages were stimulated with FM1 in the lungs of influenza A virus, respectively, with dif- ferent concentrations of 0. 388 mg/mL, 0. 194 mg/mL, 0. 097 mg/mL, 0. 048 5 mg/mL and 0. 024 25 mg/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 HIN1 virus was decreased in 0. 024 25 mg/mL group （P〈0.01）. The level of TNF-α was significantly lower in the group of Qingtouxiere De- coction 0. 097 mg/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. 048 5 mg/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 HIN1 virus group （P〈0.01）. The expression of TLR-7 protein in HIN1 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 inthe H1N1 infected group was significantly higher than that in the control group （P〈0.01）. CONCLUSION The activation of TLR-7/MyD88/NFκB signaling pathway induced by H1N1 influenza virus can inhibit the expression of downstream inflammatory factors and play the antiviral effect.