急性运动轴索型神经病患者血清对培养脊髓运动神经元的影响-比较蛋白质组研究  

Comparative proteomics study of the effects of serum from AMAN patients on the motor neurons culture in vitro

作　　者：杨国锋[1] 纪建国[2] 魏晓姗[1] 李培[1] 赵景茹[1] 张锐利[1] 冯晓[1] 

机构地区：[1]河北医科大学第二医院神经科,河北石家庄050000 [2]北京大学生命科学学院蛋白质工程国家重点实验室,北京100871

出　　处：《中风与神经疾病杂志》2010年第10期871-874,共4页Journal of Apoplexy and Nervous Diseases

摘　　要：目的为深入理解急性运动轴索型神经病(acute motor axonal neuropathy,AMAN)分子机制,以蛋白质组技术比较AMAN患者与正常对照组血清对培养的大鼠脊髓运动神经元的影响。方法原代培养脊髓运动神经元蛋白质以固相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 Master2D Elite分析电泳图谱,MALDI-TOF/TOF串联质谱鉴定在AMAN患者血清的影响下,培养的神经元发生变化的蛋白质。结果 AMAN患者血清干预24h的脊髓运动神经元与正常对照组血清干预24h的脊髓运动神经元相比较,有13个蛋白的表达量降低,8个蛋白的表达量升高,这些蛋白与能为疾病血清中致病因子作用的靶蛋白。结论在急性运动轴索型神经病患者血清作用下,培养大鼠脊髓运动神经元的细胞凋亡相关蛋白、氧化应激蛋白、突触可塑性相关蛋白、细胞骨架蛋白表达发生变化,这些发现有助于我们深入理解该疾病的病理机制。通过进一步的研究,本文发现的差异表达蛋白质也可能对急性运动轴索型神经病的诊断治疗有一定的帮助。Objective To investigate molecular mechanisms of acute motor axonal neuropathy（AMAN）,proteomics was applied to evaluate the effect of GBS patients serum on spinal motor neuron of rats.Methods The proteins extracted with mixture of urea,CHAPS,DTT,IPG buffer and protease inhibitors were run immobilized pH gradient （IPG） 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 analysised with ImageMaster 2D Elite software.The proteins of interest were in-gel digested and identified using MALDI-TOF/TOF tandem mass spectrometry.Results We found 21 differently expressed proteins including cytoskeleton proteins,apoptosis related proteins and antioxidants proteins.They were identified as heterogeneous nuclear ribonucleoprotein H,tubulin,actin Delta（3,5）-Delta（2,4）-dienoyl-CoA isomerase,proteasome subunit alpha type 1,phosphatidylethanolamine-binding protein 1,14-3-3 protein epsilon,peptidyl-prolyl cis-trans isomerase A,prohibitin,endoplasmic reticulum protein ERp29 precursor,peroxiredoxin-6,60kDa heat shock protein,F-actin-capping proteins,hypoxanthine-guanine phosphoribosyltransferase,Ras-related protein Rab-7a,triosephosphate isomerase,the receptor for activated protein kinase C 1,glyceraldehyde-3-phosphate dehydrogenase,voltage-dependent anion-selective channel protein 1,stress-70 protein,and heat shock cognate 71 kDa protein.Conclusions We got a number of related-proteins of AMAN.Some of the proteins are quite useful for discovering the molecular mechanisms of AMAN,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.

关 键 词：急性运动轴索型神经病 蛋白质组学 血清 细胞模型 

分 类 号：R744[医药卫生—神经病学与精神病学]