Ding Xiaoman, Lu Jiahai, Yu Ruoxi, Wang Xin, Wang Ting, Dong Fangyuan, Peng Bo, Wu Weihua, Liu Hui, Geng Yijie, Zhang Renli, Ma Hanwu, Cheng Jinquan, Yu Muhua, Fang Shisong
School of Public Health, Sun Yat-sen University, Guangzhou, China.
Southern Medical University, Guangzhou, China.
PLoS One. 2016 May 25;11(5):e0156017. doi: 10.1371/journal.pone.0156017. eCollection 2016.
A newly emerged H7N9 influenza virus poses high risk to human beings. However, the pathogenic mechanism of the virus remains unclear. The temporal response of primary human alveolar adenocarcinoma epithelial cells (A549) infected with H7N9 influenza virus and H1N1 influenza A virus (H1N1, pdm09) were evaluated using the proteomics approaches (2D-DIGE combined with MALDI-TOF-MS/MS) at 24, 48 and 72 hours post of the infection (hpi). There were 11, 12 and 33 proteins with significant different expressions (P<0.05) at 24, 48 and 72hpi, especially F-actin-capping protein subunit alpha-1 (CAPZA1), Ornithine aminotransferase (OAT), Poly(rC)-binding protein 1 (PCBP1), Eukaryotic translation initiation factor 5A-1 (EIF5A) and Platelet-activating factor acetylhydrolaseⅠb subunit beta (PAFAH1B2) were validated by western-blot analysis. The functional analysis revealed that the differential proteins in A549 cells involved in regulating cytopathic effect. Among them, the down-regulation of CAPZA1, OAT, PCBP1, EIF5A are related to the death of cells infected by H7N9 influenza virus. This is the first time show that the down-regulation of PAFAH1B2 is related to the later clinical symptoms of patients infected by H7N9 influenza virus. These findings may improve our understanding of pathogenic mechanism of H7N9 influenza virus in proteomics.
一种新出现的H7N9流感病毒对人类构成了高风险。然而,该病毒的致病机制仍不清楚。采用蛋白质组学方法(二维差异凝胶电泳结合基质辅助激光解吸电离飞行时间串联质谱),在感染后24、48和72小时评估了感染H7N9流感病毒和甲型H1N1流感病毒(H1N1,pdm09)的原代人肺泡腺癌上皮细胞(A549)的时间反应。在感染后24、48和72小时分别有11、12和33种蛋白质表达存在显著差异(P<0.05),尤其是通过蛋白质免疫印迹分析验证了肌动蛋白封端蛋白α-1亚基(CAPZA1)、鸟氨酸转氨酶(OAT)、聚(rC)结合蛋白1(PCBP1)、真核翻译起始因子5A-1(EIF5A)和血小板活化因子乙酰水解酶Ⅰb亚基β(PAFAH1B2)。功能分析表明,A549细胞中的差异蛋白参与调节细胞病变效应。其中,CAPZA1、OAT、PCBP1、EIF5A的下调与H7N9流感病毒感染细胞的死亡有关。这是首次表明PAFAH1B2的下调与H7N9流感病毒感染患者的后期临床症状有关。这些发现可能会增进我们在蛋白质组学方面对H7N9流感病毒致病机制的理解。