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通过建立H7N9和H1N1流感病毒(H1N1pdm09)感染人肺癌上皮细胞(A549)模型,研究病毒感染细胞后细胞蛋白质组学差异变化,探讨H7N9流感病毒感染人类致病机制。将感染复数(MOI)为0.001的H7N9、H1N1pdm09流感病毒感染A549细胞24h、48h、72h后提取细胞总蛋白进行荧光双向差异凝胶电泳(2D-DIGE)和基质辅助激光解析串联飞行时间质谱(MALDI-TOF-MS/MS)分析鉴定差异蛋白。质谱共鉴定出H7N9和H1N1pdm09流感病毒感染A549细胞24h、48h、72h上调或下调的差异蛋白分别为11、12、33个。对差异蛋白进行功能分析发现与H1N1pdm09感染组相比,(纤)丝状肌动蛋白成帽蛋白α1(F-actin-capping protein subunit alpha-1,CapZ-α1)、鸟氨酸氨基转移酶(Ornithine aminotransferase,OAT)、Poly(rC)-binding protein 1(PCBP1)、真核翻译起始因子5A-1(Eukaryotic translation initiation factor 5A-1,eIF5A)在H7N9感染A549细胞后表达量的下调加速了致细胞病变效应。血小板活化因子乙酰水解酶Ⅰb亚基β(Platelet-activating factor acetylhydrolaseIb subunit beta,PAFAH1B2)在H7N9感染A549细胞后期表达量显著降低可能与该病毒感染患者的临床症状相关。
H7N9 and H1N1 influenza virus (H1N1pdm09) were used to infect human lung cancer epithelial cells (A549) to study the changes of cell proteome in virus-infected cells and to explore the mechanism of H7N9 influenza virus infection in human. A549 cells were infected with H7N9 and H1N1pdm09 influenza viruses of infection multiplicity (MOI) of 0.001 at 24, 48, and 72 hours respectively. Total cellular protein was extracted for 2D-DIGE and MALDI-TOF-MS. -TOF-MS / MS) to identify differentially expressed proteins. Mass spectrometry identified H7N9 and H1N1pdm09 influenza virus A549 cells infected 24h, 48h, 72h up or down the differential proteins were 11,12,33. The functional analysis of the differentially expressed proteins showed that compared with the H1N1pdm09-infected group, the fibroin-like actin subunit α-1 (CapZ-α1), ornithine aminotransferase The down-regulation of expression of eukaryotic translation initiation factor 5A-1 (eIF5A) after H7N9 infection in A549 cells was accelerated by increasing the expression of poly (rC) -binding protein 1 (PCBP1) and Ornithine aminotransferase (OAT) Cytopathic effect. The decreased expression of Platelet-activating factor acetylhydrolase Ib subunit beta (PAFAH1B2) in the late stage of H7N9-infected A549 cells may be related to the clinical symptoms of the virus-infected patients.