定量蛋白质组学技术分析猪繁殖与呼吸综合征病毒感染肺泡巨噬细胞的差异表达蛋白质  

Analysis of Differential Expression Proteins in Porcine Reproductive and Respiratory Syndrome Virus Infected Pulmonary Alveolar Macrophage by Quantitative Proteomics

作　　者：刘龙[1] 陈红英[1] 

机构地区：[1]西北农林科技大学生命科学学院,杨凌712100

出　　处：《畜牧兽医学报》2016年第6期1222-1231,共10页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：中央高校基本科研业务费创新重点项目(QN2011065)

摘　　要：作者采用定量蛋白质组学方法,对猪繁殖与呼吸综合征病毒(PRRSV)感染猪肺泡巨噬细胞(PAMs)和正常PAMs的蛋白质组进行差异分析。通过高效液相色谱和电喷雾串联质谱仪定量分析PRRSV感染细胞与未感染对照细胞间的差异表达蛋白质,并运用Max-Quant 1.0.7.4软件结合蛋白质数据库PaxDb进行鉴定。与对照组相比,感染组共鉴定出39个差异表达蛋白质,其中30个蛋白质表达上调,9个蛋白质表达下调。通过Western blot和ELISA验证了感染的PAMs中SPP1、IFIT3、STAT3及IL-8的表达情况与质谱鉴定结果一致。通过液相色谱—串联质谱联用(LC-MS/MS)技术,筛选得到多个与PRRSV感染相关的差异表达蛋白质,为PRRSV发病的分子机制分析提供了新依据。The purpose of our research was to study the differential expression proteins of Porcine reproductive and respiratory syndrome virus（PRRSV）infected porcine alveolar macrophage（PAMs）relative to the un-infected control PAMs.Differential proteins in response to PRRSV infection were analyzed by High Performance Liquid Chromatography coupled with tandem mass spectrometry,and then identified using software Max-Quant 1.0.7.4combining with the protein database PaxDb.Thirty-nine differential proteins were identified in the PRRSV infected group,including thirty up-regulated proteins and nine down-regulated proteins.The up-regulation of SPP1,IFIT3 and IL-8,and down-regulation of STAT3 in PRRSV infected PAMs were verified by Western blot and ELISA.This study provides new evidences for further studies on the molecular mechanism of PRRSV pathogenesis by using LC-MS/MS.

关 键 词：猪繁殖与呼吸综合征病毒 猪肺泡巨噬细胞 差异表达蛋白质 感染 

分 类 号：S858.285.3[农业科学—临床兽医学]