蛋白质组学技术鉴定吉兰-巴雷综合征患者与正常人血清的差异表达蛋白

Proteomics study of the differential expression proteins in serum between Guillain-Barré syndrome and normal

目的为深入理解吉兰-巴雷综合征(Guillain-Barré syndrome,GBS)分子机制,以蛋白质组技术比较GBS患者与正常对照组血清的差异表达蛋白。方法血清蛋白质以固相pH梯度等电聚焦为第一向,SDS-PAGE垂直电泳为第二向进行双向电泳,图像分析软件Image Master2DElite分析电泳图谱,MALDI-TOF/TOF串联质谱鉴定差异表达蛋白。结果 24种蛋白质的表达量在吉兰-巴雷综合征患者与正常对照组显著不同,其中17种在吉兰-巴雷综合征患者血清中表达量上调的蛋白质被鉴定为补体C3、α-2-巨球蛋白、补体因子B、血浆铜蓝蛋白、补体因子H、补体C4-B、维生素D结合蛋白、血清淀粉样P物质、丛生蛋白、α-1-抗胰凝乳蛋白酶、触珠蛋白、Afamin、血红素蛋白、视黄醇结合蛋白4、补体C1s亚成分、补体C4-A、α-2锌糖蛋白;7种在吉兰-巴雷综合征患者血清中表达量下调的蛋白质被鉴定为α-1-抗胰蛋白酶、Igγ-1-C链、Igκ-C链、CD5类抗原、载脂蛋白E、血清铁传递蛋白、维生素D结合蛋白。结论 GBS患者血清中补体蛋白及急性期反应蛋白表达量发生了明显变化,这些结果加深了我们对GBS发病分子机制的理解。

Objective To investigate molecular mechanisms of Guillain-Barre＇ syndrome （GBS）,comparative proteomic analysis of serum proteins was employed to study 30 GBS patients compared to 30 controls.Methods The proteins extracted with mixture of urea,CHAPS,DTT,IPG buffer and protease inhibitors were run immobilized pH gradient isoelectric focusing electrophoresis as the first dimension,and then run vertical SDS-PAGE as the second dimension.The maps were visualized by silver staining or colloidal coomassive blue and analysed with ImageMaster 2D Elite software.The proteins of interest were in-gel digested and identified using MALDI-TOF/TOF tandem mass spectrometry.Results All 24 protein spots were differentially expressed as compared with age-matched control serum,And 17 up-regulated proteins in GBS were identified as Complement C3,Alpha-2-macroglobulin,Complement factor B,Ceruloplasmin,Complement factor H,Complement C4-B,Vitamin D-binding protein,Serum amyloid P-component,Clusterin,Alpha-1-antichymotrypsin,Haptoglobin,Afamin,Hemopexin,Retinol-binding protein 4,Complement C1s subcomponent,Complement C4-A,Zinc-alpha-2-glycoprotein.7 down-regulated proteins in GBS were identified as Alpha-1-antitrypsin,Ig gamma-1 chain C region,Ig kappa chain C region,CD5 antigen-like,Apolipoprotein E,Serotransferrin,Vitamin D-binding protein.Conclusions We got a number of related-proteins of GBS.Some of the proteins are quite useful for discovering the molecular mechanisms of GBS,and some of the proteins could be identified as disease-related protein markers to assist in diagnose and be selected as potential targets for specific drug therapy.