大鼠局灶性脑缺血脑组织的比较蛋白质组学研究.docVIP

大鼠局灶性脑缺血脑组织的比较蛋白质组学研究 王继生，邱宗荫*，夏永鹏，李惠芝 （重庆医科大学药学院 重庆 400016）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。关键词 脑缺血 蛋白质组学 A Comparative proteome study on focal cerebral ischemic tissue of rat brain Wang Ji-Sheng, 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) [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. [KEY WORDS]： cerebral ischemi