大鼠肝移植(LTx)急性排斥反应(AR)血清蛋白质组学分析  

Serum proteomic analysis of acute rat liver allograft rejection

作　　者：胡进武[1] 徐敏[1] 谭长军[1] 代智[1] 王征[1] 周俭[1,2] 樊嘉[1,2] 黄晓武[1] 

机构地区：[1]复旦大学附属中山医院肝癌研究所,上海200032 [2]上海市器官移植重点实验室-复旦大学附属中山医院,上海200032

出　　处：《复旦学报（医学版）》2014年第4期427-434,共8页Fudan University Journal of Medical Sciences

基　　金：国家自然科学基金(30700815;81272724;81225019);国家自然科学基金重大项目(81030038);国家重大科技专项项目(2012ZX10002-016);上海市自然科学基金(13ZR1406900)~~

摘　　要：目的探究肝移植(liver transplantation,LTx)急性排斥反应(acute rejection,AR)血清蛋白质组学的变化,寻找与排斥反应相关的血清标志物。方法建立Lewis→BN大鼠肝移植模型,以BN→BN大鼠肝移植为对照组,根据肝移植物AR的反应程度进行分组,取各组大鼠血清样本进行二维双向凝胶电泳(two-dimensional difference gel electrophoresis,2D-DIGE)及质谱(mass spectrometry,MS)分析,鉴定表达有差异的蛋白质。结果肝移植模型大鼠分为无排斥、轻度、中度和重度排斥4组,2D-DIGE共发现30个蛋白斑点,用MS分析排除重复蛋白斑点之后鉴定出14个蛋白质,根据功能可分为4类。第1类为与金属离子代谢相关的免疫调节蛋白,包括血红素结合蛋白、触珠蛋白前体、srprb Ba1-667和preproapo A-I;第2类为先天性免疫相关的蛋白成分,包括补体C3、补体C4a、MBP-A、LOC500183蛋白和LOC299458蛋白;第3类为免疫调节多肽,包括T-kininogen 2、α-1巨球蛋白和大鼠α-1巨球蛋白受体结合域A链晶体结构;第4类为未分类蛋白,包括ORF2和rCG36664。结论肝移植AR过程中血清多种蛋白质的含量发生改变,可能为重要的血清标志物。Objective To explore serum proteomic changes during the acute rejection （AR） after liver transplantation,and to seek the serum biomarkers associated with the AR.Methods A rat LTx model of acute allograft rejection was constructed from Lewis to Brown Norway （BN） rat,while BN to BN rats LTx served as syngeneic control.According to the severity of the AR,the recipients were grouped.Serum samples were taken from each group of recipients,and two-dimensional difference gel electrophoresis （2D-DIGE） and mass spectrometry （MS） analysis were performed to identify proteins displaying significant changes in abundance.Results The recipients were categorized into groups of no rejection,mild,moderate and severe rejection.Expressions of 30 protein spots were significantly changed.Ultimately,14 proteins were identified after the exclusion of repetitive proteins and classified into 4 types on the basis of their functions.Type 1 was metabolic immunoregulators,including hemopexin,preprohaptoglobin,Srprb Ba1-667 and preproapo A-I; Type 2 was components related to innate immune response,containing C3 complement,C4a complement,mannose-binding protein,LOC500183 protein and LOC299458 protein; Type 3 was immunoregulatory polypeptides,including T-kininogen 2,alpha-1-macroglobin and chain A crystal structure of rat alpha 1-M receptor binding domain; Type 4 was other proteins,including ORF2 and rCG36664.Conclusions The serum level of multiple proteins changes during AR after LTx,which may be important serum biomarkers.

关 键 词：肝移植(LTx) 急性排斥反应(AR) 二维双向凝胶电泳(2D-DIGE) 质谱(MS) 

分 类 号：R392.4[医药卫生—免疫学]