急性重型脑外伤后脑蛋白表达变化的蛋白质组研究

The research of acute protein expression change after severe traumatic brain injury with proteome technique

目的应用蛋白质组学技术,研究重型人脑外伤后的脑皮层蛋白质组表达变化的情况。方法提取脑挫伤部位皮层的总蛋白。通过双向电泳,分离蛋白。应用胶内酶切、生物质谱,鉴定由图像分析软件所得出的具有表达差异的蛋白质点。结果在急性期8h内,目前已发现138个蛋白质点,表达水平具有显著性差异变化。在已鉴定出的83个蛋白点中,属于64种蛋白质。依其功能可分为:细胞骨架、代谢反应、核酸蛋白合成与更新、信号传导、氧化应激反应、功能未知等几类。在急性期,大多数差异蛋白表达水平呈波动变化。结论蛋白质组技术作为一个有力的大规模、高通量的分析蛋白质混合物工具,可快速的建立脑蛋白表达谱。在急性期,脑蛋白质表达呈剧烈变化主要为参与细胞代谢反应、结构修复等组织代偿反应。

Objective To elevate the acute protein expressions level of human traumatic brain injury with proteomics technique. Methods 11 appropriate injured samples were selected in this study, the total proteins were extractel in all of them. The total proteins mixture extracted in protein lysate was separated with two dimensional gel electrophoresis. The differential protein spots analyzed by ImageMaster 5.0 software were identified by matrix associated matrix-assisted laser desorption-time of flight mass spectrometry (MALDI-TOF MS). Results After analyzing 2-D images with software, we found 138 protein spots showed significant expression levels change. 83 protein spots identified by mass spectroscopy belonged to 64 products of protein, which were divided into several functional group: cytoskeleton, intermediary metabolism, protein synthesis and turnover, and unknown function etc. Conclusions The proteomics technique including 2D-electrophoresis and MS provide us a high-throughout tool to research the global protein in brain. The human cortical proteome map provides an analytical reference for the protein expression in the brain area in health and disease. In acute phase of post-trauma, the significant changes happen in the impaired brain area, especially intermediary metabolism enzyme proteins.