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目的:研究疏风宣肺方和解表清里方体外对人肺腺癌上皮细胞A549中TLR3/7信号通路的影响。方法:利用基因芯片技术研究甲型流感病毒H1N1感染A549细胞后TLR3/7信号通路中基因转录的变化。采用荧光定量PCR和Western blotting法检测各组细胞中的Toll样受体3(Toll-like receptor3,TLR3)、Toll样受体7(Toll-like receptor 7,TLR7)、髓样分化因子88(myeloid differentiation factor 88,My D88)、激活核因子Kappa B(nuclear factor-Kappa B,NF-κB)的mRNA及蛋白表达的变化。结果:在TLR3/7相关信号传导通路中,与细胞对照组相比,H1N1感染组差异表达基因Tlr3、Tlr7、Myd88、Nfbk1、Mapk8、Mapk13、Ifna1、Ifnβ1明显上调,奥司他韦组、疏风宣肺组和解表清里组对差异表达基因Tlr3、Tlr7、Myd88、Nfbk1、Mapk8、Mapk13、Ifna1、Ifnβ1明显下调。qRT-PCR结果显示,与细胞对照组比较,H1N1感染组TLR3/7、My D88、NF-κB的mRNA表达均显著升高(P<0.01)。与H1N1感染组比较,疏风宣肺组的TLR3/7、My D88、NF-κB的mRNA表达均明显降低(P<0.05或P<0.01),解表清里方对TLR3/7、NF-κB的mRNA表达均明显降低(P<0.05或P<0.01)。同时,疏风宣肺组治疗效果优于解表清里组。Western blotting结果显示,H1N1感染组TLR3/7、NF-κB蛋白较正常组显著升高(P<0.05)。与H1N1感染组比较,两种方药的TLR3/7、NF-κB表达均显著降低(P<0.05或P<0.01)。结论:疏风宣肺方和解表清里方可以抑制流感病毒H1N1所诱导的TLR3/7信号通路活化,下调活性NF-κB的转录活性,发挥抗流感病毒的作用。
OBJECTIVE: To study the effect of Shufeng Xuanfei Fanghexibiaoqingliufang on TLR3 / 7 signal transduction in human lung adenocarcinoma A549 cells in vitro. Methods: The gene transcription in the TLR3 / 7 signaling pathway of A549 cells infected by influenza A virus H1N1 was studied by gene chip technique. The expression of Toll-like receptor 3 (TLR3), Toll-like receptor 7 (TLR7), myeloid 88 differentiation factor 88, My D88) and the activation of nuclear factor-kappa B (NF-κB) mRNA and protein expression. Results: Compared with the cell control group, the expression of Tlr3, Tlr7, Myd88, Nfbk1, Mapk8, Mapk13, Ifna1 and Ifnβ1 were significantly up-regulated in the H1N1 infected group compared with the cell control group in the TLR3 / 7 related signal transduction pathway. Wind Xuanfei group reconciliation group Qingliu group significantly downregulated differentially expressed genes Tlr3, Tlr7, Myd88, Nfbk1, Mapk8, Mapk13, Ifna1, Ifnβ1. The results of qRT-PCR showed that the mRNA expressions of TLR3 / 7, My D88 and NF-κB in H1N1 infection group were significantly increased compared with the cell control group (P <0.01). The expression of TLR3 / 7, My D88 and NF-κB in Shufeng Xuanfei group were significantly lower than those in H1N1 infection group (P <0.05 or P <0.01) MRNA expression were significantly decreased (P <0.05 or P <0.01). At the same time, Shufeng Xuanfei group is better than Xieqingli group. The results of Western blotting showed that the expression of TLR3 / 7 and NF-κB in the H1N1 group was significantly higher than that in the normal group (P <0.05). Compared with H1N1 infection group, TLR3 / 7 and NF-κB expression of two prescriptions were significantly decreased (P <0.05 or P <0.01). Conclusion: Shufeng Xuanfei Fang and Jiedu Qingrefang can inhibit the activation of TLR3 / 7 signaling pathway induced by influenza virus H1N1 and down-regulate the transcriptional activity of NF-κB, and play an anti-influenza virus effect.