疏风宣肺方和解表清里方对流感病毒H1N1感染人肺腺癌上皮细胞A549中TLR3/7信号通路作用的研究  

Effect of Shufeng Xuanfei Formula and Jiebiao Qingli Formula on TLR3 /7 Signal Pathway in Human Pulmonary Carcinoma Cell A549 and Its Mechanism

作　　者：葛世杰[1] 卢娜娜[1] 刘晓婷[1] 张沂[1] 顾立刚[1] 吴珺[1] 邱泽计[1] 张洪春[2] 晁恩祥[2] 

机构地区：[1]北京中医药大学基础医学院中医药抗病毒重点实验室,北京100029 [2]北京中日友好医院,北京100029

出　　处：《中医药学报》2015年第2期23-26,共4页Acta Chinese Medicine and Pharmacology

基　　金：国家自然科学基金资助项目(81173371)

摘　　要：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 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.

关 键 词：疏风宣肺方 解表清里方 流感病毒 人肺腺癌A549细胞 TLR3/7信号通路 

分 类 号：R285.5[医药卫生—中药学]