H7N9禽流感病毒感染A549细胞生物标志物研究

丁小满; 王婷; 彭博; 胡鹏威; 段永翔; 房师松; 俞慕华

doi:10.11847/zgggws1114088

H7N9禽流感病毒感染A549细胞生物标志物研究

A potential biomarker of influenza H7N9 virus infection in A549 cells

摘要

摘要:
目的通过建立H7N9禽流感病毒和H1N1pdm09流感病毒（对照病毒）感染A549细胞模型，分析H7N9禽流感病毒感染A549细胞的特征蛋白，筛选其生物标志物，初步揭示H7N9禽流感病毒致细胞病变的蛋白分子机制。
方法感染复数为0.001的H7N9和对照病毒分别感染A549细胞24、48、72 h后提取细胞总蛋白进行荧光双向差异凝胶电泳（2D-DIGE），筛选H7N9禽流感病毒感染A549细胞的可能生物标志物并进行蛋白印迹（WB）验证。
结果血小板活化因子乙酰水解酶Ⅰb亚基β（PAFAH1B2）蛋白在H7N9感染A549细胞72 h后表达量急剧下降，而在空白对照组和H1N1pdm09感染A549细胞组的表达量未出现明显变化。
结论 PAFAH1B2可作为H7N9感染A549细胞后期的生物标志物，其表达量的变化可能与H7N9感染患者的症状相关。

Abstract:
Objective To analyze specific protein and screen biomarkers of H7N9 avian influenza virus infection in A549 cells by establishing a A549 cell model of H7N9 avian influenza virus and H1N1pdm09 influenza virus (the control) infection for exploring the pathogenic mechanism of H7N9 avian influenza virus infection.
Methods The A549 cells were infected by H7N9 avian influenza virus with the multiplicity of infection of 0.001 and the control virus. Then the total protein of the infected cells was extracted and analyzed with two-dimensional difference gel electrophoresis (2D-DIGE) 24, 48, and 72 hours after the infection. Potential biomarkers of H7N9 avian influenza virus infection were screened and validated via Western blot.
Results Compared with that in the H1N1pdm09 infection, the platelet-activating factor acetylhydrolaseⅠb subunit beta (PAFAH1B2) was significantly down-regulated in A549 cells 72 hours after the H7N9 avian influenza virus infection. But the expression of PAFAH1B2 showed no obvious variation in A549 cells of the blank control and infected by H1N1pdm09 influenza virus.
Conclusion PAFAH1B2 could be a biomarker for later period H7N9 avian influenza virus infection in A549 cells and the variation in PAFAH1B2 expression may be associated with clinical symptoms of H7N9 avian influenza virus infection.

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