脊髓慢性压迫减压后缺血再灌注损伤的蛋白质组学研究  

Proteome analysis in ischemia-reperfusion injury after decompression of chronic spinal cord compression

作　　者：华凯[1] 郭庆升[1] 张善勇[1] 

机构地区：[1]辽宁省人民医院骨科,沈阳市110016

出　　处：《中华全科医学》2015年第6期875-877,1037,共3页Chinese Journal of General Practice

基　　金：2011年辽宁省科学技术基金项目(201102104)

摘　　要：目的研究兔颈髓慢性压迫减压后脊髓组织中差异表达蛋白质的变化,探讨其在缺血再灌注损伤中的作用机制。方法将24只兔随机分为减压组(D组)和对照组(C组),建立慢性脊髓压迫模型,减压组在减压6 h后给予致命量的戊巴比妥钠,取出颈5椎体所对应的脊髓组织,对照组致死后直接取出颈5椎体所对应的脊髓组织,进行蛋白制备和荧光染料标记,利用双向荧光差异凝胶电泳(2D-DIGE)进行蛋白质分离,以DeCyder图像分析软件对DIGE图像进行分析找到差异蛋白质位点,以基质辅助激光解吸附电离串联飞行时间质谱仪(MALDITOF/TOF)进行质谱鉴定并检索相关数据库,对蛋白质组的变化进行分析。结果减压后脊髓损伤节段蛋白质表达有明显差异,在15个差异点中共鉴定出10种蛋白质。其中下调表达蛋白3个,分别为醛糖还原酶、胞质内顺乌头酸合成酶、醛脱氢酶。上调表达蛋白7个,分别为热休克蛋白70、蛋白质二硫键异构酶3前体、α1缝隙连接蛋白、水通道蛋白4、N-乙基马来酰胺敏感因子附着蛋白、生长相关蛋白43、神经丝蛋白M。它们分别涉及到神经元的能量代谢、应激反应、细胞连接、物质转运、神经再生等功能。结论这些蛋白质可能在脊髓慢性压迫减压后缺血再灌注损伤中起作用,为今后治疗脊髓缺血再灌注损伤提供新的思路。Objective To investigate the proteomics in ischemia-reperfusion injury（IRI） after decompression of the chro- nic cervical spinal cord compression in rabbits and the role of these proteins. Methods A total of 24 rabbits were divided into decompression group（ Group D） and control group（ Group C ）. An experimental animal model of chronic spinal cord compression was established. After the decompression of six hours in Group D,lethal amounts of sodium pentobarbital was given to both groups, and the spinal cord in C5 vertebral body was taken out to prepare the proteins and label with fluoro- chrome;two-dimensional fluorescence difference gel electrophoresis （2D-DIGE） was performed for separating proteins and DeCyder image analysis software was used for analyzing the image of D1GE, then matrix-assisted laser desorption ionization tandem time of flight mass spectrometer（MALDI-TOF/TOF） was used to identify the mass spectrum. Finally the proteins with different expression were analyzed through searching proteins database. Results There were significant differences in protein expression of the injured lesions of spinal cords before and after the decompression. And 10 kinds of differential proteins were identified from a total of 15 differential protein spots, of which 3 were down-regulated（ including aldose re- ductase, cytoplasmic aconitate hydratase and aldehyde dehydrogenase） and 7 were up-regulated（ including heat shock 70, protein disulfide isomerase-associated 3 precursor, gap junction alpha-1 protein, aquaporins 4, N-ethylmaleimide-sensitive factor attachment protein, growth associated protein 43, neurofilament proteins M） in Group D. They were involved in neu- ronal energy metabolism, stress, cell connection, material transport, nerve regeneration and other functions. Conclusion These identified proteins may play an important role in IRI after decompression of the chronic spinal cord compression.

关 键 词：脊髓 缺血再灌注损伤 动物实验 蛋白质组学 

分 类 号：R651.2[医药卫生—外科学] R338.21[医药卫生—临床医学]