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目的:研究毒热平注射液对流感病毒感染的小鼠腹腔巨噬细胞株Ana-1细胞TIR(Toll/IL-1 receptors)信号传导通路的影响。方法:用100 TCID50流感病毒亚甲型鼠肺适应株A/FM/1/47(H1N1)感染小鼠腹腔巨噬细胞株Ana-1后换用10,1 mg.L-1含药维持液继续培养,分别于12,24 h弃细胞上清液并收集细胞,提取巨噬细胞RNA,进行实时定量PCR反应(RT-PCR),动态测定毒热平注射液对病毒感染前后巨噬细胞TIR信号传导通路中各信号蛋白:Toll样受体7(TLR7)、髓样分化因子88(MyD88),IL-1受体相关激酶4(IRAK4)、肿瘤坏死因子相关激酶6(TRAF6)和核因子κB(NF-κB)mRNA表达水平的影响。结果:毒热平注射液可剂量依赖性地下调病毒感染巨噬细胞TLR7,MyD88,IRAK4,TRAF6,NF-κB mRNA的表达水平。结论:毒热平注射液能通过调节TIR信号传导通路各信号蛋白的活性发挥抗病毒感染作用。
OBJECTIVE: To study the effect of Fureiping injection on TLR (Toll/IL-1 receptors) signaling pathway in influenza virus-infected mouse macrophage cell line Ana-1. METHODS: Infection of mouse peritoneal macrophage cell line Ana-1 with 100 TCID50 influenza virus subtype mouse lung adapted strain A/FM/1/47 (H1N1) was replaced with 10,1 mg.L-1 drug-containing maintenance solution. After incubation, cell supernatants were discarded at 12, 24 h and cells were harvested. Macrophage RNA was extracted and real-time quantitative PCR (RT-PCR) was performed to dynamically determine the TIR of macrophage cells before and after viral infection with Fulingping Injection. Signaling pathways in signaling proteins: Toll-like receptor 7 (TLR7), myeloid differentiation factor 88 (MyD88), IL-1 receptor-associated kinase 4 (IRAK4), tumor necrosis factor-associated kinase 6 (TRAF6), and nuclear factor Effect of NF-κB mRNA expression level. RESULTS: DPP could down-regulate the expression levels of TLR7, MyD88, IRAK4, TRAF6 and NF-κB mRNA in virus-infected macrophages in a dose-dependent manner. Conclusion: Tofuping injection can exert antiviral effect by regulating the activity of each signal protein in TIR signaling pathway.