论文部分内容阅读
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。
Aims: To investigate the effect of Shufeng Xuanfei Recipe and Xiebailingli Recipe on the apoptosis of A549 cells infected with influenza virus H1N1. Methods: Human lung adenocarcinoma A549 cells and A549 cells were infected with influenza A virus H1N1 and then treated with Tamiflu as drug control group. The signal transduction pathway of apoptosis was analyzed by KEGG pat hway assay using gene chip technique. To study the changes of gene transcription in the signal path of apoptosis in Shufeng Xuanfei Fanghexibiaoqingliufang and the expression of Fas ligand (Fas), Fas ligand (Fas) in each group of cells by fluorescence quantitative PCR L) and caspase-3, -8, -9 (Caspase-3, -8, -9) m RNA expression were detected by Western blotting assay Changes in the protein in the cell. Results: Compared with the cell control group, the expression of Casp3, Casp8, Casp9, Fas and Fasl were significantly up-regulated in H1N1 infection group. The expression of Casp8, Casp7, The results of q RT-PCR showed that the m RNA expression of Caspase-3, -8, -9, Fas and Fas L were significantly increased in H1N1 infection group compared with that in control group (P <0.01); Compared with H1N1 infection group, m RNA expression of Caspase-3, -8, -9, Fas and Fas L were significantly decreased in Shufeng Xuanfei group (P <0.05) The m RNA expression of Fas and Fas L decreased significantly (P <0.05). At the same time Shufeng Xuanfei group treatment better than the solution in the clear group. The results of Westen blot analysis showed that the number of Fas and Fas L protein in H1N1 infection group was significantly higher than that in control group (P <0.05). Compared with H1N1 infection group, the expressions of Fas and Fas L were significantly decreased in both prescriptions (P <0.05). The result of q RT-PCR was basically consistent with that of gene chip. CONCLUSIONS: Shufeng Xuanfei Fang and Xiedu Qingliufang can regulate the expression of Caspase-3, -8, -9, Fas and Fas L after influenza A virus infection, which can resist the apoptosis after influenza virus infection.