慢重型乙肝患者血浆蛋白质组差异表达的初步研究  

Differential expression of plasma proteomics in severe chronic hepatitis B

作　　者：欧强[1] 彭霞[1] 殷科珊[1] 张丽军[1] 

机构地区：[1]上海市（复旦大学附属）公共卫生临床中心感染科,上海201508

出　　处：《公共卫生与临床医学》2009年第4期260-263,279,共5页Public health and dinical medicine

基　　金：上海市卫生局青年课题（2007041）;中国肝炎防治基金会血液净化人工肝攻关课题（200704）

摘　　要：目的探讨慢性重型乙肝病变的血浆蛋白质组变化，为研究慢性重型乙肝的发病机制及治疗提供新的线索。方法收集健康对照者，慢性重型乙肝患者血浆各10份，去除血浆中高丰度蛋白。应用双向凝胶电泳（2-DE）技术构建两组研究对象血浆蛋白2-DE图谱，经ImageMaster差异分析软件进行分析，寻找差异蛋白质点。用电喷雾离子串联质谱（ESI-MS—MS）对重要的差异蛋白质点进行鉴定。结果经过样品预处理后，血浆中白蛋白和免疫球蛋白IgG含量大大减少，低丰度蛋白得到较好的富集。ImageMaster 2D Platinum6．0软件将银染图谱分成2个类别进行找点、匹配等分析，正常健康组蛋白质点287个，慢性重型乙肝组蛋白质点297个，筛选出2倍以上的差异蛋白质点31个。对差异蛋白质点进行ESI—MS—MS分析，鉴定了8个慢性重型乙肝相关蛋白质，其中上调的蛋白质有α1-抗胰蛋白酶，载脂蛋白E；下调的蛋白质有补体因子B；CD5抗原相关蛋白；纤维蛋白原B链；α-1B糖蛋白；载脂蛋白AI；触珠蛋白。结论慢性重型乙肝患者血浆蛋白质组与正常人血浆对比存在明显差异，成功鉴定了8个慢性重型乙肝相关蛋白。这些差异蛋白质的功能涉及能量代谢、脂蛋白代谢、蛋白质分泌、补体旁路活化途径等过程。对这些重要蛋白质的结构和功能的进一步研究，对于进一步揭示慢性重型乙肝发病分子机制及寻找治疗靶标可能具有重要的意义。Objective To characterize plasma protein profile during severe chronic hepatitis B and to provide new clues for exploring the molecular mechanism and the new therapeutic strategies. Methods High abundance proteins were removed from plasmas which were collected from severe chronic hepatitis B （SCHB） patients and health control. The left plasma were separated by two-dimensionai electrophoresis. The gels were stained by silver nitrate or Coomassie blue and analyzed by Imagemaster software. The differently expressed proteins were identified by electrospray ionization mass spectrometry （ESI-MS-MS） . Results Through pretreatment, the high abundance proteins were largely removed and the low abundance proteins were enriched. More than 280 protein spots were separated in Coomassie blue stained gels. 31 proteins with two-fold difference were observed through Imagemaster software analysis. 8 proteins were identified as SCHB specific proteins, in which Alpha-1-antitrypsin precursor and Apolipoprotein E precursor were up-regulated, and CD5 antigen-like precursor, Fibrinogen beta chain precursor, Alpha-1B-glycoprotein precursor, Apolipoprotein A-I precursor, Haptoglobin precursor were down-regulated. Conclusions Protein profile of plasma from severe chronic hepatitis B patients was different from control; 8 proteins that are engaged in energy metabolism , lipoprotein metabolism and complement bypass were identified SCHB specific proteins. These observations have important implications for future research on the molecular mechanism and new therapeutic strategies for severe chronic hepatitis B.

关 键 词：慢性重型乙肝 血浆 蛋白质组学 双向凝胶电泳 质谱 

分 类 号：R512.62[医药卫生—内科学] R446.1[医药卫生—临床医学]