摘要
目的比较海马注射淀粉样蛋白β(Aβ)大鼠与正常大鼠脑蛋白质组双向电泳(2-DE)图谱差异,从蛋白质水平初步探索阿尔茨海默病(AD)发病机制。方法以固相pH梯度等电聚焦为第1向,SDS-PAGE垂直电泳为第2向进行2-DE。以图像分析软件ImageMaster 2D-Elite分析电泳图谱。结果海马注射Aβ大鼠与正常大鼠脑组织2-DE图谱分别检出496和491个蛋白点。对2张电泳图进行匹配后,发现有11个蛋白点仅在海马注射Aβ大鼠脑蛋白2-DE图谱中表达,而有6个蛋白点只在正常对照大鼠检测到。部分蛋白在2组大鼠脑组织中含量发生了明显变化。结论初步建立了AD动物模型比较蛋白质组学的技术方法;差异点的发现为深入理解AD发病机制及研发新药提供了有益的线索。
Objective To investigate the molecular mechanisms of Alzheimer's disease by comparing global protein patterns in two-dimensional electrophoregram (2-DE) of the brain of rats with intrahippocampal amyloid β injection and normal rats. Methods From adult SD rats with intrahippocampal injection of amyloid β, 200 μg brain proteins were extracted with 9 mol/L urea, 4% CHAPS, 1% DTT, 0.5% CA and a cocktail of protease inhibitors. Immobilized pH gradient (IPG) isoelectric focusing electrophoresis of the extracted proteins was performed to obtain the electrophoretogram of the first dimension, with the second dimension obtained by vertical SDS-PAGE. The electrophoretograms were visualized using silver staining and analyzed with ImageMaster 2D-Elite software. Results On average, 496 and 491 protein spots could be obtained in the electrophoregraphs for rats with amyloid β and the control rats, respectively, and 30 of these spots exhibited quantitative changes. Another 11 and 6 spots were exclusively shown on the protein maps for amyloid β-treated rats and control rats, respectively. Conclusion The differentially displayed proteins in the brain identified between the rats with intrahippocampal amyloid β injection and control rats may provide further insight into the pathogenesis of Alzheimer's disease and useful clues for developing new drugs for its treatment.
出处
《第一军医大学学报》
CSCD
北大核心
2004年第5期553-555,共3页
Journal of First Military Medical University
