双向电泳—质谱法探索实验性重症肌无力大鼠疾病相关蛋白  

Mass Spectrometric Proteome Analyses of Serum Obtained From Lewis Rats Suffering From Experimental Autoimmune Myasthenia Gravis

作　　者：刘萍[1] 丁旭峰[1] 章怡祎[1] 乔健[2] 

机构地区：[1]上海中医药大学附属龙华医院,上海200032 [2]复旦大学附属华山医院,上海200040

出　　处：《中华中医药学刊》2007年第12期2486-2489,共4页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(30472204)

摘　　要：目的:分离和鉴定实验性自身免疫性重症肌无力大鼠血清差异表达蛋白,寻找并比较疾病相关蛋白,以进一步阐明重症肌无力的发病机制。方法:采用二维蛋白电泳技术分离正常组和模型组大鼠血清差异表达蛋白,凝胶经银染显色后,Bio-Rad凝胶扫描仪扫描,Imagemaster图像软件分析,差异蛋白点用基质辅助激光解析电离-两级飞行时间串联质谱法进行鉴定。结果:通过双向电泳,建立血清蛋白质双向电泳图谱,重症肌无力组血清双向电泳图谱可见678个蛋白点,而正常对照组可见702个蛋白点。选取9个具有明显差异的蛋白点进行质谱鉴定,共鉴定出5种蛋白质。结论:正常组和模型组大鼠间存在一定的差异表达蛋白,该类蛋白为一些与机体免疫应答、信息调控相关的蛋白质。Objective ： To separate and identify the different serum - associated proteins of experimental autoimmune myasthenia gravis（EAMG） rats for understanding the pathogenesis of EAMG. Methods： The total proteins from either EAMG rats or normal ones were prepared by means of two-dimensional gel eleetrophoresis. After silver staining and apparatus scanning, the analysis of gel -image was performed by using Imagemaster. Then the differentially expressed proteins were identified by matrix assisted laser desorption time - of - flight time - of - flight mass spectrometry （ MALDI - TOF - TOF）. Results ： The two - dimensional polyacrylamide gel electrophoresis profiles of serum proteins were successful established by 2 - DE. The 2 - DE map of myasthenia gravls rats showed that 678 protein spots were visualized, while 702 protein spots were visualized in normal control ones. Ten of the significant differential proteins were selected and identified by MALDI - TOF - TOF. Five of them was finally identified which were correlated to immunoreaction, formulation, etc. Conclusion： The differentially expressed proteins can be observed in serum from EAMG and controls, which can be used to elucidate the pathogenesis of EAMG for further study.

关 键 词：重症肌无力 蛋白质组学 双向电泳 血清 差异蛋白 

分 类 号：R-33[医药卫生]