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大鼠局灶性脑缺血脑组织的比较蛋白质组学研究

A COMPARATIVE PROTEOME STUDY ON FOCAL CEREBRAL ISCHEMIC TISSUE OF RAT BRAIN

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【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。

【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.

【关键词】 脑缺血蛋白质组学
【Key words】 cerebral ischemicProteomics
【基金】 绵阳市科委资助课题(编号:06S042-7)
  • 【会议录名称】 2008年中国药学会学术年会暨第八届中国药师周论文集
  • 【会议名称】2008年中国药学会学术年会暨第八届中国药师周
  • 【会议时间】2008-10
  • 【会议地点】中国河北石家庄
  • 【分类号】R743
  • 【主办单位】中国药学会、河北省人民政府
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