脑出血与脑缺血模型大鼠脾淋巴细胞的蛋白质组学研究  

Proteomic Analysis of Splenolymphocyte in Rat With Intracerebral Hemorrhage and Cerebral Ischemia

作　　者：熊新贵[1] 梁清华[1] 陈疆[2] 曾年菊[1] 区健刚[1] 范荣[1] 

机构地区：[1]中南大学湘雅医院中西结合研究所,长沙410008 [2]中南大学湘雅医院远程医疗中心,长沙410008

出　　处：《中国生物工程杂志》2008年第9期7-13,共7页China Biotechnology

基　　金：国家自然科学基金重大研究计划(90409002);湖南省自然科学基金(06JJ20012)资助项目

摘　　要：目的:通过比较正常与脑出血及脑缺血模型大鼠脾淋巴细胞蛋白质表达的差异,初步探讨细胞免疫功能与脑血管病之间的关系。方法:将SD大鼠随机分为正常组、脑出血模型组(采用VII型胶原酶诱导脑出血)和局灶性脑缺血模型组(采用线栓法造成大脑中动脉阻塞),分离大鼠脾淋巴细胞,提取总蛋白质后进行双向凝胶电泳,考马斯亮蓝染色,PDQUEST软件分析,对差异蛋白质点采用基质辅助激光解析电离质谱(MALDI-TOF-MS)技术进行鉴定并分析。结果:胶质细胞成熟因子γ等9个蛋白在脑出血和脑缺血模型组表达上调,膜联蛋白III在脑出血和脑缺血模型组表达下调。结论:建立了分辨率高重复性较好的脑出血及局灶性脑缺血脾淋巴细胞总蛋白的双向凝胶电泳图谱,并鉴定一些与脑血管病脑损伤相关的差异表达蛋白质,为深入研究脑血管病细胞免疫功能改变与脑血管病之间的关系奠定了基础。To primarily approach the correlation between cellular immune function and cerebrovascular disease（CVD） injury by analyzing the difference of protein expression in normal and intracerebral hemorrhage （ICH） as well as middle cerebral artery occlusion （MACO） rats of splenolymphocyte. Methods： SD rats were randomly divided into normal group, intracerebral hemorrhage group （intracerebral hemorrhage induced by Ⅶ- collagenase） and focal cerebral infarction group （middle cerebral artery occlusion made by a thread ）. The splenolymphocyte of rats were isolated and the total proteins were extracted and separated by two-dimensional polyacrylamide gel electrophpresis （2-DE） which was followed by Coomassie Brilliant Blue staining to visualize the protein spots in the 2-DE gels . The differentially expressed proteins were analyzed using PDQUEST analysis software,and then identified by peptide mass fingerprint （PMF） based on matrix- assisted laser desorption/ ionization time of flight mass spectrometry （MALDI-TOF-MS）. Results： Several proteins were up-regulated including Glia maturation factor gamma , Rho, GDP dissociation inhibitor （GDI） beta , Peroxiredoxin-Ⅱ , Hypothetical protein, Beta-actin, NADH dehydrogenase （ubiquinone） flavoprotein 2,60S acidic ribosomal protein P0 and 40S ribosomal protein SA and annexin Ⅲ was down-regulated in rat with. intracerebral hemorrhage and cerebral ischemia. The differentially expressed proteins were able to be divided into the following several groups based on their functions ： proteins related to signal tranduction, antioxidant, protein biosynthesis, metabolic enzymes,structual proteins et al. Conclusions：We obtained the 2-DE pattern with high resolution and better repeatability of splenolymphocyte in rat with cerebral hemorrhage and cerebral ischemia, subsequently, ten differentially expressed proteins were preliminarily identified. This approach may lay a foundation for the further investigation to the relationship between cellular im

关 键 词：蛋白质组学 动物模型 脑出血 脑缺血 脾淋巴细胞 

分 类 号：TQ93[化学工程]