【作者】 王继生； 张芳； 邱宗荫； 李惠芝； 周成林；

【Author】 Wang Ji-Sheng1,Zhang-fang,Qiu Zong-Yin2,Li Hui-Zhi2,Zhou cheng-lin (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:采用蛋白质组学技术对尼莫地平对脑缺血再灌注大鼠脑组织差异蛋白进行研究,进一步揭示尼莫地平的脑保护作用及抗损伤修复机制。方法:建立脑缺血再灌注大鼠动物模型,分别提取蛋白质,2-DE电泳分离,应用相关软件比较分析,找出差异蛋白,酶切消化后进行肽指纹图谱分析,数据检索,初步鉴定差异蛋白质。结果:局灶性脑缺血模型组与尼莫地平干预组的比较蛋白质组学研究。与模型组比较,从尼莫地平干预组找到了25个差异表达蛋白斑点,其中10个低表达,15个高表达,鉴定其中6个蛋白质,发现激肽原-1前体、谷氨酸受体4前体等与脑缺血相关的蛋白质。结论:尼莫地平对脑缺血再灌注的保护作用是多个蛋白共同参与的结果。更多还原

【Abstract】 Objective:With comparative proteomics strategy and technique,The purpose of the research was to screen alteration in protein level in brain tissue between middle cererbral artery occlusion(MCAO) and nimodipine treated rats. to learn the mechanism of nerve-protecting effects of nimodipine after focal cerebral ischemia-reperfusion. Methods:We established the MCAO 2h reperfusion of 24h model in rats, 2-DE was applied to preparein step 1 for the separation of protein, imaging analysis of gels was done with Image-Master 2D(V2002.1)software. Different protein spots expressing between cerebral ischemic reperfusion (CIR)model and nimodipine group were cut from the gels for the MS analysis, Different protein spots expressing between cerebral ischemic tissue administered with nimodipine and not-administered were also cut from the gels for the MS analysis. MALDI-TOF-MS and database search:protein spots from step 2 were subject to in-gel digestion and MALDI-TOF-MS analysis, which resulted in the peptident mass fingerprint spectra(PMF). PMF were search against swiss-prot database using search program of Ms-Fit. Results:A comparative proteome study of model and normal group .Compared with model group, the normal group gained 25 differential protein spots,15 spots expressed lowly ,and 10 spots high., identified 6 protein spots,found out the relative cerebral ischemic proteins such as Kininogen-1 precursor and Glutamate receptor 4 precursor etc. Conclusion:Nimodipine on cerebral ischemia-reperfusion is the protective effect of the participation of a number of protein results.更多还原