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为进一步了解流感病毒(IFZ)是否像呼吸道合胞病毒(RSV)一样在感染过程中可以上调呼吸道上皮细胞粘蛋白1(MUC1)的表达,进而限制炎症的发展。我们利用qRT-PCR和Western Blot检测两种呼吸道单负链RNA病毒感染人呼吸道上皮细胞系——A549细胞后对MUC1的表达调控。分别利用人喉上皮细胞系——HEp-2细胞系和MDCK细胞系培养并收集RSV和IFZ。相同滴度的两种病毒分别感染A549细胞24h后,分别裂解各组细胞,收集总RNA,应用qRT-PCR检测MUC1mRNA的表达情况;或相同滴度的两种病毒分别感染A549细胞24h和48h后,裂解细胞收集总蛋白,同时收集细胞培养上清,应用Western Blot检测MUC1蛋白的表达情况,应用ELISA检测上清中TNF-α水平。结果显示,虽然两种病毒都能上调TNF-α水平,但只有RSV可以上调呼吸道上皮细胞MUC1表达,并呈剂量效应,而IFZ不能上调呼吸道上皮细胞MUC1的表达。本研究首次探讨两种常见呼吸道单负链RNA病毒感染呼吸道上皮细胞后对MUC1表达调控的差异,初步证实临床上IFZ感染后病情自限性的机制不同于RSV感染,与MUC1的表达上调无关。
To further understand whether influenza virus (IFZ) can upregulate the expression of respiratory epithelial mucin 1 (MUC1) during infection, like respiratory syncytial virus (RSV), thereby limiting the development of inflammation. We used qRT-PCR and Western Blot to detect the expression of MUC1 in two human respiratory tract epithelial cell line-A549 cells after infection with two negative respiratory RNA viruses. RSV and IFZ were cultured and collected using human laryngeal epithelial cell line -HEp-2 cell line and MDCK cell line, respectively. A549 cells were infected with the same titer of the two viruses respectively 24h later, the cells were lysed and the total RNA was collected. The expression of MUC1 mRNA was detected by qRT-PCR or A549 cells were infected with the same titer for 24h and 48h respectively , Lysed cells collected total protein, while collecting cell culture supernatant, the application of Western Blot detection MUC1 protein expression, the application of ELISA detection of supernatant TNF-α levels. The results showed that although both viruses could upregulate the level of TNF-α, only RSV could up-regulate MUC1 expression in respiratory epithelial cells in a dose-dependent manner, whereas IFZ failed to up-regulate the expression of MUC1 in airway epithelial cells. This study is the first to investigate the difference of MUC1 expression between two common respiratory negative single-stranded RNA viruses infected with respiratory epithelial cells. It is preliminary confirmed that the mechanism of disease self-limiting in IFZ infection is different from RSV infection and has no relation with MUC1 up-regulation.