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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细胞核因子-κB(NF-κB)活性的影响。方法:用100TCID50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10mg/L和1mg/L浓度含药维持液继续培养,分别于12h和24h弃细胞上清液并收集细胞,提取巨噬细胞RNA和核蛋白,进行实时定量PCR反应(RT-PCR)和Western blot实验,动态测定毒热平注射液对病毒感染前后巨噬细胞Toll样受体7(TLR7)、髓样分化因子88(MyD88)mRNA和NF-κBp65mRNA及其蛋白表达水平的影响。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、MyD88mRNA的表达、NF-κBp65mRNA及其蛋白表达水平。结论:毒热平注射液能通过调节MyD88依赖的信号传导通路下调细胞核因子-κB活性发挥抗病毒感染作用。
Objective: To study the effect of Tachienping Injection on the activity of nuclear factor-κB (NF-κB) in mouse peritoneal macrophage cell line Ana-1 infected with influenza virus. Methods: The murine peritoneal macrophage cell line Ana-1 was infected with 100TCID50 influenza A virus-adapted lung strain A / FM / 1/47 (H1N1) and then switched to 10mg / L and 1mg / L drug- The cells were harvested and the cells were harvested at 12 h and 24 h, respectively. Macrophages RNA and nuclear proteins were extracted, and real-time quantitative PCR (RT-PCR) and Western blot were performed to determine the effect of Tachien-Ping injection on viral infection (TLR7), myeloid differentiation factor 88 (MyD88) mRNA and NF-κBp65 mRNA and protein levels in macrophages. Results: Tachikuiping injection can down-regulate the expression of TLR7, MyD88 mRNA and the expression of NF-κBp65 in macrophages infected by virus in a dose-dependent manner. Conclusion: TDP can exert anti-viral infection by regulating MyD88-dependent signal transduction pathway to downregulate nuclear factor-κB activity.