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首页> 中文期刊> 《中医药学报》 >疏风宣肺方和解表清里方对流感病毒 H1 N1感染人肺腺癌上皮细胞 A549中 TLR3／7信号通路作用的研究

疏风宣肺方和解表清里方对流感病毒 H1 N1感染人肺腺癌上皮细胞 A549中 TLR3／7信号通路作用的研究

         
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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H1 N1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 receptor 3，TLR3）、Toll样受体7（Toll－like receptor 7，TLR7）、髓样分化因子88（myeloid differentiation factor 88， MyD88）、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、Nf-bk1、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、MyD88、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、MyD88、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， H1 N1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H1 N1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 investigate the regulatory effects of Shufeng Xuanfei and Jiebiao Qingli Chinese medicine formu-lae on TLR3/7 signal pathway in human pulmonary carcinoma cell A549 .Methods: Gene chips were used to screen these RNA samples in virus-infected A549.Select differentially expressed genes of TLR3/7 signal pathway.The mRNA and protein expressions of TLR3/7, MyD88, NF-κB were verified by Real-Time PCR and western-blot.Results:Tlr3, Tlr7, Myd88, Nfbk1, Mapk8, Mapk13, Ifna1 and Ifnβ1 were up-regulated in virus-infected group.Shufeng Xuanfei and Jiebiao Qingli formulae could down -regulate gene expressions of Tlr3, Tlr7, Myd88, Nfbk1, Mapk8, Mapk13, Ifna1 and Ifnβ1.Real-Time PCR and western blot experiments showed that two formulae could significantly decrease mRNA and protein expressions of TLR3/7, MyD88 and NF-κB (P<0.01), compared with the virus-infec-ted group.The results also figured that the interference efficacy of Shufeng Xuanfei formula was better than that of Jiebi-ao Qingli.As was expected, real-time PCR and western-blot data were in agreement with the microarray assay.Con-clusion:Shufeng Xuanfei and Jiebiao Qingli formulae can down-regulate the activity of NF-κB by regulating MyD88 in TLR3/7 signal pathway, thus fighting against influenza virus in vitro.

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