疏风宣肺方与解表清里方对流感病毒H_1N_1感染的A549细胞凋亡的影响  

Influence of ShuFeng XuanFei Formula and JieBiao QingLi Formula on the Apoptosis of A549 Cells Infected by Influenza Virus H_1N_1

作　　者：葛世杰[1] 刘晓婷[1] 张沂[1] 卢娜娜[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

出　　处：《西部中医药》2015年第5期7-10,共4页Western Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(编号81173371)

摘　　要：目的:探讨疏风宣肺方与解表清里方对流感病毒H1N1感染的A549细胞凋亡的影响。方法:培养人肺腺癌上皮细胞A549,甲型流感病毒H1N1感染A549细胞后,以达菲为药物对照组,利用基因芯片技术通过KEGG pat hway分析涉及细胞凋亡信号传导途径。研究疏风宣肺方与解表清里方凋亡信号通路中基因转录的变化;同时采用荧光定量PCR检测各组细胞中的肿瘤坏死因子受体超家族成员6(Fas)、Fas配体(Fas L)及天门冬氨酸特异性半胱氨酸蛋白-3、-8、-9(Caspase-3、-8、-9)m RNA表达的变化;West er n bl ot t i ng法检测各组细胞中蛋白的变化。结果:与细胞对照组相比,H1N1感染组差异表达基因Casp3、Casp8、Casp9、Fas、Fasl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Casp8、Casp7、Fas、Fasl明显下调;q RT-PCR结果显示,与细胞对照组比较,H1N1感染组Caspase-3、-8、-9、Fas、Fas L的m RNA表达均显著升高(P<0.01);与H1N1感染组比较,疏风宣肺组Caspase-3、-8、-9、Fas、Fas L的m RNA表达均明显降低(P<0.05),解表清里组Caspas e-3、-9、Fas、Fas L的m RNA表达均明显降低(P<0.05)。同时疏风宣肺组治疗效果优于解表清里组。Wes t er n bl ot t i ng结果显示,H1N1感染组Fas、Fas L蛋白细胞较对照组显著升高(P<0.05)。与H1N1感染组比较,2种方药的Fas、Fas L表达均显著降低(P<0.05);q RT-PCR结果与基因芯片结果基本一致。结论:疏风宣肺方与解表清里方均可调控甲型流感病毒感染后的Caspase-3、-8、-9、Fas、Fas L表达,可对抗流感病毒感染后的细胞凋亡。Objective：To explore the influence of ShuFeng XuanFei formula and JieBiao QingLi formula on the apoptosis of A549 cells induced by influenza virus H1N1 infection. Methods：After human lung adenocarcinoma epithelial cells A549 was cultured and infected by influenza A H1N1, Tamiflu was taken as the control group of medicine, Cellular apoptosis signal transduction pathway was involved in the analysis of KEGG pathway by using microarray technology. microarray analysis was used to study the changes of genetic transcription in apoptosis sig-naling pathway regulated by both formulas;at the same time, realtime fluorescence quantitative PCR was adopted to detect the expressions of tumor necrosis factor receptor （TNFR） super family members 6 （Fas）, Fas ligand （FasL）, Caspase-3,-8,-9 mRNA in the cells of different groups;Western blotting method was applied to observe the change of the protein in the cells. Results：Compared with the control group of the cells, differential genes expressions of Casp3, Casp8, Casp9, Fas and FasL increased obviously in H1N1 infection group, while Casp8, Casp7, Fas and FasL decreased significantly in oseltamivir group, ShuFeng XuanFei formula group and JieBiao QingLi formula group;qRT-PCR results showed that, compared with the control group of the cells, mRNA expressions of Casp3, Casp8, Casp9, Fas and FasL in H1N1 infection group improved notably （P〈0.01）;compared with H1N1 infection group, the mRNA expressions of Casp3, Casp8, Casp9, Fas and FasL in ShuFeng XuanFei formula group reduced remarkably （P〈0.05）, while the mRNA expressions of Casp3, Casp9, Fas and FasL in JieBiao QingLi formula group significant-ly（P〈0.05）. Curative effects of ShuFeng XuanFei formula group were better than that of JieBiao QingLi formula group. Western blotting results indicated that H1N1 infection group was improved more notable than the control group in Fas and FasL albuminous cell（all P〈0.05）. Compared with H1N1 infection group, the expressions of Fas and FasL decreased in

关 键 词：流感病毒 人肺腺癌A549细胞 细胞凋亡 疏风宣肺方 解表清里方 

分 类 号：R186[医药卫生—流行病学]