应用8标iTRAQ技术结合2D LC-MS/MS分析大鼠再生肝的差异蛋白质组  

Differential Proteomic Analysis in Rat Liver Regeneration Using 8-Plex iTRAQ Labeling Coupled with 2D LC-MS/MS

作　　者：张一甫[1] 秦兆宇[1] 刘晓慧[2] 林成招[1] 贺福初[1,3] 

机构地区：[1]复旦大学生物医学研究院系统生物学实验室,上海200032 [2]复旦大学生物医学研究院蛋白质组研究中心,上海200032 [3]北京放射医学研究所北京蛋白质组研究中心蛋白质组学国家重点实验室,北京102206

出　　处：《分析测试学报》2010年第5期421-429,共9页Journal of Instrumental Analysis

基　　金：国家自然科学基金资助项目(30900257);上海市自然科学基金资助项目(09ZR1404100);北京蛋白质组学国家重点实验室自主课题资助项目(SKLP-K200808)

摘　　要：首次应用8标iTRAQ(Isobaric tags for relative and absolute quantitation)技术结合2D LC-MS/MS(Two-dimensional liquid chromatography-tandem mass spectrometry),以部分肝切除的大鼠为模型,全面分析了再生早期(6、12 h)、中期(24、48 h)和晚期(72、120 h)肝脏蛋白质的表达差异。共鉴定357种蛋白质,6个时间点共有60种蛋白质的表达量与对照组(手术切下的正常肝组织)有显著差异。其中,ADH1(Alcohol dehy-drogenase 1)和PRDX1(Peroxiredoxin 1)等的表达变化趋势与已有报道相同,GO(Gene ontology)的蛋白质功能注释提示ADH1、CAT(Catalase)、ENO1(Enolase 1)和APOA1(Apolipoprotein A-Ⅰ)等可能通过影响细胞凋亡和能量供给等方式参与肝再生调控。聚类分析表明,术后12 h和24 h蛋白质的整体表达模式相似,6 h与此二者相近,48 h和72 h相似,而120 h与这些时间点相差最远。该研究验证了8标iTRAQ是并行处理多组样本的有效技术,适合于生理和病理过程多时间点蛋白质组的定量差异分析,可以监控关键分子的动态变化,更易发掘重要的调控靶点分子。该文为进一步探讨肝再生机理提供了有价值的线索。A comparative proteomic analysis in rat liver regeneration after partial hepatectomy was firstly performed using 8-plex iTRAQ（Isobaric tags for relative and absolute quantitation） technology coupled with 2D LC-MS/MS（Two-dimensional liquid chromatography-tandem mass spectrometry）.Differentially expressed proteins in the early stage（6 h,12 h）,middle stage（24 h,48 h） and late stage（72 h,120 h） were profiled.Among 357 distinct proteins identified,60 proteins were up-or down-regulated with significance at one time point or more.ADH1（Alcohol dehydrogenase 1）and PRDX1（Peroxiredoxin 1）had similar expression patterns as previous reports.The outcome from functional analysis with GO（Gene ontology） annotation system suggested that ADH1,CAT（Catalase）,ENO1（Enolase 1） and APOA1（Apolipoprotein A-Ⅰ）,affecting apoptosis and glycolysis during regeneration,might be the regulation targets.Hierarchical clustering analysis showed that proteins expression patterns of 6 h was similar to those of 12 h and 24 h,that of 48 h similar to 72 h,and of 120 h distinct from all other time points.The work demonstrated that 8-plex iTRAQ was an ideal technology for comparative proteomic study of multiple samples,which could figure out a dynamically quantitative expression pattern of proteins.In addition,the results also provided some useful clues for further research on mechanism of liver regeneration.

关 键 词：ITRAQ 2D LC-MS/MS 肝脏再生 差异蛋白质组 

分 类 号：O657.63[理学—分析化学] O629.73[理学—化学]