文档介绍：大鼠局灶性脑缺血脑组织的比较蛋白质组学研究
王继生1,邱宗荫* *绵阳市科委资助课题(编号:06S042-7)
作者简介:王继生(1974~),男,药理学博士,研究方向:新药开发的药理学研究及蛋白质组学,Email:wjs327@
2,夏永鹏2,李惠芝2
(1绵阳市第三人民医院绵阳 621000;2 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。
[关键词] 脑缺血蛋白质组学
PARATIVE PROTEOME STUDY ON FOCAL CEREBRAL ISCHEMIC TISSUE OF RAT BRAIN
Wang Ji-Sheng1, Qiu Zong-Yin2, Xia Yong-Peng2, Li Hui-Zhi2 ( 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 protein
s 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 :.parative 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 : Establishing a 2-DE technology was applied to protein analysis of brain tissue ,and found out th