急性脑外伤后的比较蛋白质组学研究

Comparative proteome analysis of human cortex protein expression after severe traumatic brain injury

目的 研究脑损伤前后人脑皮层蛋白质组分差异表达的情况。方法对10例重型脑外伤患者和4例对照的皮层组织进行比较蛋白质组学研究，即利用固相pH梯度（immobilized pH gradient，IPG）双向凝胶电泳（two—dimensional electrophoresis，2-DE）分离人脑外伤后皮层组织及对照的皮层组织的总蛋白质，蓝银显色后经图像分析识别差异表达的蛋白质，应用基质辅助激光解吸电离飞行时间质谱（matrix—assisted laserdesorption／ionization time—of—flightmass spectrometry，MALDI—TOF—MS）和电喷雾-四极杆-飞行时间串联质谱（electrospray ionization—quadrupole time—of—flight，ESI—Q—TOF）鉴定差异表达的蛋白质点。结果得到了分辨率较高、重复性较好的人脑外伤后皮层组织及对照的皮层组织的2-DE图谱；图像分析识别差异蛋白质点21个；通过质谱分析鉴定出一些与代谢反应、氧化应激反应、信号转导等有关的差异蛋白17个。结论成功鉴定17个与脑外伤相关的蛋白，为进一步阐明脑外伤后的病理生理作用机制及其诊断和治疗、预后监测提供了新的思路。

Objective To explore differential proteomic expression of human traumatic brain cortex tissue before and after severe traumatic brain injury （TBI）. Methods Comparative proteome analysis was conducted on brain cortex tissues from 10 cases with severe TBI and 4 normal controls. The total proteins of human traumatic cortex tissues and paired normal tissue were separated by means of immobilized pH gradient （IPG） -based two-dimensional gel electrophoresis （2-DE） and silver staining. The differential protein expression was analyzed and then identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry （MALDI-TOF-MS） and electrospray ionization-quadrupole time-offlight MS/MS（ESI-Q-TOF）. Results Well-resolved reproducible 2-DE patterns of human traumatic cortex tissues and normal ones were obtained. Twenty-one differentially expressed proteins were found by image analysis. Seventeen differential proteins were identified by mass spectroscopy, with some being the products of metabolic reaction or oxidative stress while others involving the regulation of cell cycle and signal transduction. Conclusion A total of 17 proteins associated with TBI are identified in this study, which provides a new clue for researches on post-traumatic mechanism, diagnosis, treatment and prognosis.