大鼠局灶性脑缺血脑组织的比较蛋白质组学研究

目的建立局灶性脑缺血模型,从蛋白质整体水平探讨脑缺血/再灌注的损伤机制。方法建立大鼠脑缺血2 h再灌注24 h的模型;提取脑组织总蛋白质,以双向电泳技术进行蛋白质分离,考马斯亮蓝染色,获得双向电泳图谱,利用专门的软件筛选出差异蛋白质,用MALDI-TOF-MS获得肽指纹图谱,MS-Fit数据库进行搜索,获得有关的蛋白质信息。结果局灶性脑缺血模型组与正常组脑组织的比较蛋白质组学研究。与模型组比较筛选出23个差异表达蛋白斑点,其中13个低表达,10个高表达,鉴定其中6个蛋白质,发现白细胞三烯A-4水解酶、促甲状腺激素释放激素降解胞外酶等与脑缺血相关的蛋白质。结论从蛋白质组学角度发现了一些与脑缺血相关的蛋白,有助于今后进一步研究脑缺血性损伤的保护机制。

Study on compared proteomics of focal cerebral ischemia in rats

Aim To establish a model of focal cerebral ischemic tissue of rat brain and explore the injury mechanism of focal cerebral ischemia reperfusion in whole level of proteins.Method The model was established with suture method by reperfusion 24 h after ischemic 2 h according to Koizumi′s method,total brain tissue proteins were extracted with Lysis buffer,proteins were separated by two-dimensional electrophoresis(2-DE),stained by Coomassie brilliant blue,the patterns were gotten,differential proteins were found out,PMS was obtained by MALDI-TOF-MS,and related information of proteins was gained by MS-Fit database.Results A comparative proteomic study of model and normal group was performed.Compared with model group,the normal group gained 23 differential protein spots,13 spots expressed lowly,and 10 spots high,6 protein spots were identified,the relative cerebral ischemic proteins such as Leukotriene A-4 hydrolase,Thyrotropin-releasing hormone-degrading ectoenzyme etc were found out.Conclusions Establishing a 2-DE technology is applied to protein analysis of brain tissue,and the relative proteins of cerebral ischemia are found from proteome aspect.This will contribute to the research on the injury mechanism of cerebral ischemic tissues.