颅脑创伤后亚低温脑保护的蛋白质组研究

The differential proteomic studies of hippocampus following traumatic brain injury in rats with mild hypothermia

目的本研究应用差异蛋白质组学技术，探讨亚低温和常温条件下，创伤性颅脑损伤大鼠海马组织蛋白质的表达变化。方法采用侧方液压冲击装置，建立大鼠中度脑损伤模型，亚低温组（n=3）于伤后维持体温（33±0．5）℃持续3h，常温组（n=3）始终维持体温（37±0．5）℃。取大鼠海马组织，通过差异荧光双向凝胶电泳、分离蛋白，获得二维的蛋白质分离图谱，然后通过胶内酶切、抽提酶解肽段、基质辅助激光解吸飞行时间质谱分析差异的蛋白质点，鉴定出变化的蛋白质。结果通过DeCyder5．0（GE Heahhcare）软件分析报告发现差异1．5倍以上的蛋白点17个（P〈0．05）。通过对这些蛋白考染点进行质谱鉴定，鉴定出14个蛋白质，2个为同一种蛋白，实际差异蛋白数为13个。分别是细胞骨架蛋白、介导能量代谢的酶类、参与核酸合成、氧化应激反应的蛋白质、神经突触功能蛋白、细胞内信号传递蛋白及未知蛋白。结论脑损伤后亚低温及常温条件下，大鼠海马组织蛋白质存在表达差异，鉴定出的差异蛋白质可能与亚低温保护效应的潜在作用机制有关。

Objective To elevate the effects of mild hypothermia on protein expression of hippocampus following traumatic brain injury in rats by differential proteomics technique. Methods The hippocampus was taken from rats （ n = 3 each group） underwent moderate fluid-percussion （F-P） brain injury with and without hypothermic treatment （33 ± 0.5）℃/3h. The total proteins mixture extracted in protein lysate was separated with two dimensional differential fluorescence in-gel electrophoresis （ DIGE ）. The differential protein spots were identified by matrix-assisted laser desorption ionization time of flight mass spectrometry（MS-TOLF）. Results Seventeen protein spots were found with statistically significant change, its Ratio value was 〉 1.5. Fourteen products of protein spots were identified by mass spectroscopy, two of them were repeated, the actual products of proteins were thirteen. They included cytoskeletons, energy-related enzymes, nucleic acids synthetic proteins, oxidative and stress proteins, synapse function related proteins, intracellular signal transduction proteins, as well as unknown proteins. Conclusion Mild hypothermia has an evident effect on different expressions of hippocampus proteins following moderate traumatic brain injury , which may be related to the protective mechanisms of mild hypothermia. The protective role of some predicted proteins on mild hyperthermia were not reported.