大鼠缺血性脑卒中脑皮层蛋白质组学分析

摘要

Objective To explore brain cortex proteomics changes during ischemic cerebral stroke (ICS),and to seek the biomarkers associated with ICS.Methods A total of 42 male SD rats were randomly divided into sham group (n =12) and ICS group (n =30).ICS model was induced by distal middle cerebral artery occlusion.Brain cortex tissues were collected at 1,6,24 and 48 h after ICS,respectively,after cerebral ischemia.Samples from different groups were subjected to tandem mass tag (TMT)-based quantitative proteomics analysis for identification of differentially expressed proteins.Gene Ontology enrichment analysis was utilized to classify the differentially expressed proteins.Western blot was used to verify the quantitative proteomics results.Results Compared with sham group,25 proteins were considered to be differentially expressed in ICS group.Gene Ontology enrichment analysis revealed that the differentially expressed proteins were related to acute-phase response,inflammatory response,lipoprotein metabolic process,complement activation,classical pathway and innate immune response.The expression change of heat shock protein 27 (HSP27) was basically consistent with the quantitative proteomics results.Conclusions These 25 differentially expressed proteins may serve as potential biomarkers for ICS.%目的 探讨缺血性脑卒中(ischemic cerebral stroke,ICS)后脑皮层蛋白质组的变化,寻找与其相关的生物标志物.方法 42只雄性SD大鼠分为假手术组(sham组,n=12)和脑缺血组(ICS组,n=30).利用远端大脑中动脉梗阻(distal middle cerebral artery occlusion,dMCAO)构建ICS模型,分别在脑缺血后1、6、24和48 h取脑皮层组织.对不同组的样品进行基于串联质谱标签(tandem mass tag,TMT)标记的定量蛋白质组学分析,鉴定表达有差异的蛋白质;利用Gene Ontology富集分析对差异表达蛋白质进行分类;利用Western blot验证定量蛋白质组学的结果.结果 与sham组比较,ICS组共鉴定到25个差异表达蛋白质.Gene Ontology富集分析显示差异表达蛋白质主要与急性期反应、炎症反应、脂蛋白代谢过程、补体激活经典途径及固有免疫反应有关.热休克蛋白27(heat shock protein 27,HSP27)的表达变化与定量蛋白质组学的结果基本一致.结论 这25个差异表达蛋白质可能作为ICS潜在的生物标志物.