Pick病的蛋白质组研究

目的为深入理解Pick病分子机制并寻找诊断治疗该疾病的蛋白质标记物，选取3例经病理证实的Pick病患者与4例正常老年人尸检脑标本进行蛋白质组学研究。方法死亡24h内新鲜取材的尸检额叶、颞叶蛋白质以固相pH梯度等电聚焦为第一向，十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）为第二向进行双向电泳（2-DE），图像分析软件Image Master 2D Elite分析电泳图谱，基质辅助激光解析／电离-飞行时间（MALDI-TOF）质谱或MALDI-TOF／TOF串联质谱鉴定蛋白质。结果15种蛋白质的表达量在Pick病患者与正常老年人显著不同。8个在Pick病表达量上调的蛋白质被鉴定为热休克蛋白60、Mn超氧化物歧化酶、α-烯醇化酶、3-磷酸甘油醛脱氢酶、胶质纤维酸性蛋白、硫氧还蛋白过氧化物酶1、RNA结合蛋白调节亚基、P25α；7个在Pick病表达量下调的蛋白质被鉴定为pemxiredoxin 5、cofilin、铁蛋白H链、血清白蛋白前体、二氢嘧啶酶相关蛋白2、丙酮酸激酶、碳酰还原酶[NADPH]1。结论氧自由基与抗氧化蛋白的平衡受损及重要代谢酶、神经原纤维缠结相关蛋白的变化同Pick病发病密切相关。

Proteomic study of Pick''''s disease

Objective To investigate the molecular mechanisms of Pick's disease.Methods A comparative proteomic analysis of brain proteins was employed to 3 patients with Pick's disease and compared to 4 controls.The brains of subjects who died without clinical or pathological involvement of nervous system and brains of Pick's disease patients were obtained at autopsy.The brain proteins extracted with mixture of urea,CHAPS,DTT,immobilized pH gradient(IPG) buffer and protease inhibitors were run by IPG isoelectric focusing electrophoresis as the first dimension,and then by run vertical SDS-PAGE as the second dimension.The maps were visualized by using silver staining or colloidal coomassive blue and analysised with Image Master 2D Elite software.The proteins of interest were in-gel digested and identified using MALDI-TOF mass spectrometry or MALDI-TOF/TOF tandem mass spectrometry.Results All 15 protein spots were differentially expressed as compared with age-matched nondemented control brains.And 8 up-regulated proteins in Pick's disease brains were identified as Heat shock 60 000 protein 1,Mn-superoxide dismutase,alpha enolase,glyceraldehyde 3-phosphate dehydrogenase,glial fibrillary acidic protein,t hioredoxin peroxidase 1,RNA-binding protein regulatory subunit,P25 alpha.And 7 down-regulated proteins in Pick's disease brains were identified as human peroxiredoxin 5,cofilin,ferritin heavy chaio,serum albumin precursor,dihydropyrimidinase-related protein 2,pyruvate kinase,carbonyl reductase 1.Conclusions It suggested that the mechanism of Pick's disease should be closely related with the damage of important metabolic pathway,the increasing of free radicals and the changes of neurofibrillary tangle(NFT)-related proteins.