荧光差异双向凝胶电泳法筛选大鼠脑缺血相关蛋白  

Cerebral ischemia related protein identified by fluorescence two-dimensional differential in-gel electrophoresis in rats

作　　者：马学玲[1] 刘亢丁[1] 江新梅[1] 文佳媚[1] 李桂玉[1] 李楠[1] 

机构地区：[1]吉林大学第一医院神经内科,吉林长春130021

出　　处：《中国老年学杂志》2007年第11期1025-1028,共4页Chinese Journal of Gerontology

基　　金：国家自然科学基金(30470588)

摘　　要：目的鉴定局灶性脑缺血相关蛋白并筛选早期神经保护蛋白。方法应用先进的差异蛋白质组学荧光差异双向凝胶电泳(2D DIGE)技术,比较大鼠大脑中动脉闭塞(MCAO)6h病灶侧大脑皮层和正常大鼠相应部位蛋白质变化;采用DeCyder-DIA软件、单因素方差分析ANO- VA,选择两组间蛋白表达量差异具有统计学意义(P<0.05)且AR>1.4的蛋白点;基质辅助激光解析/电离-飞行时间(MALDI-TOF)质谱鉴定差异蛋白。结果脑缺血6h组与正常对照组比较13个蛋白点符合统计学要求;经质谱分析仅鉴定出一个缺血组明显减少的蛋白点为α-微管蛋白。结论作为结构蛋白之一的α-微管蛋白在脑缺血早期即发生明显变化,是缺血性脑血管病早期相关蛋白。Objective To identify cerebral ischemia-relevant proteins and to screen early neuron-protection protein. Methods A comparative proteomic approach, fluorescence two-dimensional differential in-gel electrophoresis （2D DIGE） was used to compare the changes in protein between the left hemisphere cerebral cortex tissues following 6 h middle cerebral artery occlusion （MCAO） and the corresponding sites of healthy control rats. Protein spots expression levels showed statistically significant （ P〈0.05 ） and AR 〉1.4 on every gel were selected by DeCyder differential in-gel analysis （DIA） software and one-way analysis of variance （ANOVA）. Peptide mass fingerprinting was used by matrix-assisted laser desorption/ionization-time of flight （MALDI-TOF） mass spectrometry to identify all difference protein spots. Results There were 13 protein spots consistent with statistical requests in Cy3 and Cy5 labeled samples between ischemia 6 h group and control group. Only one protein was identified as α-tubulin significantly decreased in the ischemia rats. Conclusions α-tubulin as one of the cytoskeletal proteins whose expression levels are decreased in the early stage after MCAO focal ischemia, which is related proteins of cerebral ischemia at early stage.

关 键 词：蛋白质组学 差异双向凝胶电泳 Α-微管蛋白 大鼠 脑缺血 

分 类 号：Q51[生物学—生物化学] R734.31[医药卫生—肿瘤]