脑出血与脑缺血动物模型脾淋巴细胞的蛋白质组学研究

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 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。

Proteomic analysis of Splenolymphocyte in rat with cerebral hemorrhage and cerebral ischemia

ABSTRACT：Aim: To approach the relationship between cellular immune function and cerebrovascular disease(CVD) injury by estab1ish two-dimensiona1 electrophoresis(2-DE)maps of Splenolymphocyte in rat with intracerebral hemorrhage(ICH) and cerebral ischemia ,and identify the differentially expressed proteins．Methods: SD rats were randomly divided into normal group, intracerebral hemorrhage group (intracerebral hemorrhage induced by VII-collagenase and cerebral infarction group (a thread of a middle cerebral artery occlusion model). The Splenolymphocyte of rats were isolated and the total proteins were extracted and separated by 2-DE .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). Resuits：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、40S ribosomal protein SA were significantly increased, annexin III was significantly reduced in in rat with intracerebral hemorrhage and cerebral ischemia. Conclusions：The well-resolved and reproducible 2-DE pattern of Splenolymphocyte were established in rat with cerebral hemorrhage and cerebral ischemia and ten differentially expressed proteins were characterized.This approach may lay a foundation for the further investigation of the relationship between cellular immune function and cerebrovascular disease(CVD) injury.