血浆置换对慢性重型乙型肝炎患者血浆蛋白质组影响的初步研究  

Primary study on changes of plasma proteomics in chronic severe hepatitis B treated with plasma exchange

作　　者：欧强[1] 殷科珊[1] 彭霞[1] 陈良[1] 张丽军[1] 

机构地区：[1]上海市公共卫生临床中心感染科,上海201508

出　　处：《中国血液净化》2009年第9期499-502,共4页Chinese Journal of Blood Purification

基　　金：上海市卫生局青年科研项目基金资助(2007Y41);中国肝炎防治基金会血液净化人工肝攻关科研项目基金资助(SLDALBP200701)

摘　　要：目的探讨慢性重型乙型肝炎血浆置换(P E)治疗前后的血浆蛋白质组变化,为研究PE治疗重型肝炎的作用机制及寻找治疗靶标提供新的线索。方法收集慢性重型乙型肝炎患者PE前后血浆各15份,去除血浆中高丰度蛋白质。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Image Master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等高丰度蛋白质含量明显减少,低丰度蛋白得到较好的富集。PE前组蛋白质点297个,PE后组蛋白质点305个,筛选出3倍以上的差异蛋白质点15个,鉴定了5个差异蛋白质,其中在PE后血浆中上调的蛋白质有补体因子B(Complement factor B precursor,CFB,点1)、CD5antigen-like蛋白(CD5antigen-like precursor,点2)、纤维蛋白原B链(Fibrinogen beta chain precursor,FGB,点3)、触珠蛋白(Haptoglobin precursor,点4),下调的蛋白质有载脂蛋白E(Apo-E,点5)。结论鉴定了5个PE治疗前后差异表达的蛋白质,这些蛋白表达的改变提示PE可以干预机体脂蛋白代谢、蛋白质分解合成代谢,并能调节机体免疫功能,补体旁路活化途径等。深入研究这些蛋白质结构和功能对于进一步揭示PE的治疗作用机制具有重要的临床意义。Objective To study plasma protein changes in chronic severe hepatitis B after plasma exchange （PE）, to provide new clues for the understanding of PE mechanism, and to seek the new therapeutic targets for patients with chronic severe hepatitis B. Methods Plasma samples were obtained from chronic severe hepatitis B patients before and after PE. After removal of high abundanceproteins in plasma, proteins in samples were separated by two-dimensional electrophoresis. The gels were then stained by silver nitrate or Coomassie blue, and analyzed by the Imagemaster software. Proteins in the spots showing remarkable intensity difference before and after PE were identified by electrospray ionization mass spectrometry （ESI-MS/MS）. Results After the pretreatment procedures, the high abundance proteins such as albumin and IgG in the samples were mostly removed, and the low abundance proteins were enriched. From the silver stained gels, 297 protein spots from samples before PE and 305 protein spots from samples after PE were separated. From these spot, 15 proteins were found to have 3-fold differences before and after PE, and 5 proteins were identified. After PE, complement factor B precursor, CD5 antigen-like precursor, fibrinogen beta chain precursor and haptoglobin precursor were up-regulated, and apolipoprotein E precursor was down-regulated. Conclusion We have identified 5 plasma proteins showing remarkable difference in expression level before and after PE. These proteins are involved in lipoprotein metabolism, protein synthesis and catabolism, and activation of complement bypass route. They are worthwhile to be further investigated for understanding the mechanism of PE.

关 键 词：慢性重型乙型肝炎 血浆置换 蛋白质组 双向凝胶电泳 质谱 

分 类 号：R318.14[医药卫生—生物医学工程]