APP转基因小鼠与正常小鼠脑蛋白质组双向电泳图谱的比较

Comparison of two-dimensional electrophore tograms of brain proteome between APP transgenic and control mice

目的探讨APP转基因小鼠与正常C57小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老年性痴呆发病机制。方法APP转基因小鼠与正常C57小鼠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,SDS-PAGE垂直电泳为第二向进行2-DE。图像分析软件ImageMaster 2D Elite分析电泳图谱。结果APP转基因小鼠与正常小鼠脑组织2-DE图谱分别检测出976和947个蛋白点。对两张电泳图进行匹配后,发现有16个蛋白点仅在APP转基因小鼠脑蛋白2-DE图谱检测到表达,而有7个蛋白点只在正常小鼠检测到。部分蛋白在2组小鼠脑组织中含量发生了明显变化。结论差异点的发现初步建立了差异表达蛋白质组学的技术方法;为研究阿尔茨海默病机制及研发新药提供了有益的线索。

Objective To compare the global protein patterns between APP transgenic mice and control mice brains by means of two-dimensional electrophoresis （2-DE）. Methods Two hundred micrograms of mouse brain proteins were extracted with 9 mol/L urea, 4% （w/v） CHAPS and 1% （w/v） DTT and a cocktail of protease inhibitors was run through immobilized pH gradient （IPG） isoelectric focusing electrophoresis as the first dimension, and then run through vertical SDS-PAGE electrophoresis as the second dimension. The map was visualized after silver staining and analysed with ImageMaster 2D Elite software. Results 976 and 947 protein spots were obtained in APP transgenic mice and control mice brain maps respectively, of which 39 spots increased or decreased in quantity. Other 16 and 7 spots were exclusively found in APP transgenic mice and control mice brain maps, respectively. Conclusion The differentially displayed proteins between APP transgenic mice and control mice brains are quite useful for diagnosis and treatment of Alzheimer＇ s disease.