【作者】 王继生； 邱宗荫； 夏永鹏； 李惠芝；

【Author】 Wang Ji-Sheng1, Qiu Zong-Yin2, Xia Yong-Peng2, Li Hui-Zhi2 (1.The third hospital of mianyang, mianyang 621000; 2 Department of Pharmacy, ChongQing University of Medical Science. ChongQing 400016)

【机构】 绵阳市第三人民医院； 重庆医科大学药学院；

【摘要】 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/再灌注的保护作用机制。方法雄性、健康大鼠SD,随机分为正常组、脑缺血模型组参照Koizumi法并加以改进,建立MCAO大鼠脑缺血2h再灌注24h的模型;裂解液提取总蛋白质,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,从正常组筛选到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。更多还原

【Abstract】 Objective: To establish a model of focal cerebral ischemic tissue of rat brain and explore the protective mechanism of focal cerebral ischemia reperfusion in whole level of proteins. Method: Male and healthy SD rats , were classified them into normal , Model at random. Established the MCAO model with suture method by reperfusion 24h after ischemic 2h according to Koizumi,s method, extracted total proteins with Lysis buffer,separated proteins by two-dimensional electrophoresis(2-DE), stained proteins by Coomassie brilliant blue ,got the patterns ,found out differential proteins ,and obtained PMS by MALDI-TOF-MS, gained related the information of proteins by MS-Fit database.Result:.A comparative proteome study of model and normal group. Compared with model group, the normal group gained 23 differential protein spots, 13 spots expressed lowly, and 10 spots high, identified 6 protein spots,found out the relative cerebral ischemic proteins such as Leukotriene A-4 hydrolase, Thyrotropin-releasing hormone-degrading ectoenzyme etc.Conclusion: Establishing a 2-DE technology was applied to protein analysis of brain tissue ,and found out the relative proteins of from proteome aspect. This would profit from the protective mechanism of cerebral ischemic tissue.更多还原