【摘要】：通過建立H7N9和H1N1流感病毒(H1N1pdm09)感染人肺癌上皮細(xì)胞(A549)模型,研究病毒感染細(xì)胞后細(xì)胞蛋白質(zhì)組學(xué)差異變化,探討H7N9流感病毒感染人類致病機(jī)制。將感染復(fù)數(shù)(MOI)為0.001的H7N9、H1N1pdm09流感病毒感染A549細(xì)胞24h、48h、72h后提取細(xì)胞總蛋白進(jìn)行熒光雙向差異凝膠電泳(2D-DIGE)和基質(zhì)輔助激光解析串聯(lián)飛行時(shí)間質(zhì)譜(MALDI-TOF-MS/MS)分析鑒定差異蛋白。質(zhì)譜共鑒定出H7N9和H1N1pdm09流感病毒感染A549細(xì)胞24h、48h、72h上調(diào)或下調(diào)的差異蛋白分別為11、12、33個(gè)。對差異蛋白進(jìn)行功能分析發(fā)現(xiàn)與H1N1pdm09感染組相比,(纖)絲狀肌動蛋白成帽蛋白α1(F-actin-capping protein subunit alpha-1,CapZ-α1)、鳥氨酸氨基轉(zhuǎn)移酶(Ornithine aminotransferase,OAT)、Poly(rC)-binding protein 1(PCBP1)、真核翻譯起始因子5A-1(Eukaryotic translation initiation factor 5A-1,eIF5A)在H7N9感染A549細(xì)胞后表達(dá)量的下調(diào)加速了致細(xì)胞病變效應(yīng)。血小板活化因子乙酰水解酶Ⅰb亞基β(Platelet-activating factor acetylhydrolaseIb subunit beta,PAFAH1B2)在H7N9感染A549細(xì)胞后期表達(dá)量顯著降低可能與該病毒感染患者的臨床癥狀相關(guān)。
[Abstract]:By establishing the model of H7N9 and H1N1 influenza virus (H1N1pdm09) infecting human lung cancer epithelial cells (A549), we studied the proteomic changes of cells infected with H7N9 influenza virus, and discussed the pathogenesis of H7N9 influenza virus infection. The total protein was extracted from A549 cells infected with H7N9H1pdm09 influenza virus with a complex (MOI) of 0.001 for 24 h or 48 h and analyzed by fluorescence two dimensional differential gel electrophoresis (2D-DIGE) and matrix assisted laser desorption tandem time-of-flight mass spectrometry (MALDI-TOF-MS/MS). The up-regulated or down-regulated proteins of H7N9 and H1N1pdm09 influenza virus infected A549 cells for 24 h and 48 h / 72 h were 1112,33 respectively. Functional analysis of differentially expressed proteins revealed that filamentous actin cap protein 偽 1 (F-actin-capping protein subunit alpha-1,CapZ- 偽 1), ornithine aminotransferase (Ornithine aminotransferase,OAT) Poly (rC) binding protein 1 (PCBP1) and eukaryotic translation initiation factor 5A-1 (Eukaryotic translation initiation factor 5A-1e IF5A were infected with A549 cells after H7N9 infection. The down-regulation of expression accelerates the cytopathic effect. The decreased expression of platelet activating factor acetylhydrolase 鈪，

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