慢性重型乙型肝炎患者血浆蛋白质组差异表达的初步研究

The primary research on different expressing of plasma proteomics in chronic severe hepatitis B

目的探讨慢性重型乙型肝炎患者血浆蛋白质组变化,为研究慢性重型乙型肝炎的发病机制及治疗提供新的线索。方法收集慢性重型乙型肝炎患者和健康对照者血浆各20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含量大大减少,低丰度蛋白得到较好的富集。正常健康组蛋白质点287个,慢性重型乙型肝炎组蛋白质点297个,筛选出2倍以上的差异蛋白质点21个。鉴定出8个慢性重型乙型肝炎相关蛋白质,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抗原样蛋白、纤维蛋白原B链、α-1B糖蛋白、载脂蛋白AI、触珠蛋白。结论成功鉴定了8个慢性重型乙型肝炎相关蛋白质。这些差异蛋白质功能涉及能量代谢、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型肝炎发病分子机制及寻找治疗靶标可能具有重要的意义。

Objective To search for disease-associated serum proteins in chronic severe hepatitis B, and provide novel clues to identify the molecular mechanism and therapy of chronic severe hepatitis B. Methods The serum proteins from chronic severe hepatitis B patients and health control were profiled quantitatively, and two-dimensional electrophoresis （2DE） was applied to separate and compare with the pre-treated serum samples. The gels were stained by silver nitrate or Coomassie blue and analyzed by Imagemaster software. The different expressed proteins were identified by electrospray ionization mass spectrometry （ESI-MS/MS）. Results By comparing with normal and chronic severe hepatitis B serum samples, we found that at least 21 proteins were significantly changed in the sera of chronic severe hepatitis B. And 8 greatly altered proteins were identified to be Alpha 1-antitrypsin precursor and Apolipoprotein E precursor which were up-regulated, while CD5 antigen-like precursor, Fibrinogen beta chain precursor, Alpha-lB- glycoprotein precursor, Apolipoprotein A-I precursor, Haptoglohin precursor which were down-regulated.. Conclusion Protein profiles of sera from chronic severe hepatitis B patients were altered. Eight different proteins, which involve in energy metabolism, lipoprotein metabolism and complement bypass were identified. It might contribute to further research the molecular mechanism and therapy of chronic severe hepatitis B.