淀粉样β蛋白前体转基因小鼠与正常小鼠脑蛋白质组蛋白质表达差异  

Difference of proteomic protein expression in brains between amyloid protein precusor transgenic mice and normal mice

作　　者：杨国锋[1] 彭立威[1] 姚建华[1] 何思志[2] 纪建国[2] 于东华[1] 李冬梅[1] 

机构地区：[1]河北医科大学第二医院神经科,河北省石家庄市050000 [2]北京大学蛋白质工程国家重点实验室,北京市100871

出　　处：《中国临床康复》2006年第18期59-62,共4页Chinese Journal of Clinical Rehabilitation

摘　　要：目的:以蛋白质组学技术研究淀粉样β蛋白前体转基因小鼠与正常小鼠脑蛋白质表达差异,从蛋白质分子水平探索老年性痴呆发病机制。方法:实验于2002-03/2003-10在河北医科大学第二医院、解放军总医院老年医学研究所、北京大学蛋白质工程国家重点实验室完成。淀粉样β蛋白前体转基因小鼠4只与正常C57小鼠4只脑组织经蛋白提取后,分别以固相pH梯度等电聚焦为第一向,十二烷基硫酸钠-聚丙烯酰胺凝胶垂直电泳为第二向进行双向电泳。图像分析软件Im-ageMaster2DElite分析电泳图谱。以MALDI-TOF质谱仪对差异蛋白进行鉴定。结果:8只小鼠均进入结果分析。11个蛋白点在淀粉样β蛋白前体转基因小鼠与正常小鼠脑组织表达量明显不同。他们分别被鉴定为热休克蛋白86ku-1、神经丝三联体L、氧化还原酶75ku亚单位前体、血清白蛋白前体、甘油-3-磷酸脱氢酶、三磷酸腺苷合酶α亚单位、α-烯醇化酶、γ-烯醇化酶、泛素结合酶E2、肌酸激酶、天冬氨酸氨基转移酶。结论:通过蛋白质组学技术,寻找并鉴定了淀粉样β蛋白前体转基因小鼠与正常小鼠脑差异表达蛋白质。AIM： To study the difference of protein expression in brain with the proteomic technique between amyloid protein precusor （APP） transgenic mice and normal mice, and investigate the molecular mechanisms of Alzheimer disease. METHODS： The experiments were carried out in the Second Hospital of Hebei Medical University, the Institute of Geriatrics, General Hospital of Chinese PLA, and the National Key Laboratory of Protein Engineering, Peking University between March 2002 and October 2003. After brain proteins were extracted in 4 APP transgenic mice and normal mice, the brain tissues were fixed with immobilized pH gradient （IPG） isoelectric focuslng electrophoresis as the first dimension, and then run vertical odium dodecyl sulfate-pelyacrylamide gel electrophoresis （SDS-PAGE） as the second dimension. The electrophoretogram was analyzed with ImageMaster 2D Elite imaging analytical software. The proteins of interest were in-gel digested and identified with MALDI-TOF mass spectrometry or MALDITOF/TOF tandem mass spectrometry. RESULTS： All the 8 mice entered the analysis of results. Eleven protein spots were differentially expressed in APP transgenic mice as compared with the control mice brains, which were identified as heat shock protein 86 ku-1, neurofilament triplet L, oxidoreductase 75 ku subunit precursor, serum albumin precursor, glyeeraldehyde 3-phosphate dehydrogenase, ATP synthase alpha subunit, alpha enolase, gamma enolase, ubiquitin-conjugating enzyme E2, creatine kinase and aspartate transaminase. CONCLUSION： The differentially displayed proteins between APP transgenic mice and control mice brains were found and identified with the proteomic technique.

关 键 词：阿尔茨海默病 淀粉样Β蛋白前体 小鼠 转基因 淀粉样Β蛋白 电泳 凝胶 双向 蛋白质组 

分 类 号：R749.4[医药卫生—神经病学与精神病学]