【摘要】 目的建立急性脊髓损伤双向凝胶电泳图谱,初步观察急性脊髓损伤组织中蛋白质组的变化与差异表达,为进一步探讨急性脊髓损伤分子机制奠定了基础。方法采用双向凝胶电泳分离脊髓总蛋白,银染显色,Image Master图象分析系统分析,从胶中选取分离蛋白质点,基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)分析获取肽质量指纹图谱(PMF),Mascot软件搜索MSDP、SWISS-PROT数据库鉴定蛋白质。结果获得重复性和分辨率较好的脊髓双向凝胶电泳图谱。质谱分析了12个蛋白质点,获取6张肽质量指纹图谱。通过查询数据库鉴定了9个蛋白质。通过对过表达脊髓损伤中蛋白质表达谱改变的研究,鉴定出9个差异表达蛋白质,这些蛋白质包括等。这些差异蛋白质涉及基因为上调蛋白钙蛋白酶Calpain、神经丝蛋白M Neurofilament tripletα1-糖蛋白α1-Acid-glycoprotein、Oxygen regulatedprotein 1在脊髓损伤组下调。钙蛋白酶抑制剂Calpain inhibitor,假象蛋白-10 Hypothetical protein cgi-10在脊髓损伤组消失。并在损伤组出现新蛋白预测蛋白Hypothetical 152.1 kDa protein和核内寡聚肽酶Endooligopeptidase A related protein。结论成功构建脊髓损伤与正常脊髓双向凝胶电泳图谱。并应用质谱技术鉴定了特异蛋白质点。在我们所鉴定的脊髓损伤过程中发生最显著变化的蛋白质中,有7种蛋白具有与一致的变化趋势,其中钙蛋白酶、神经丝蛋白、神经营养素低亲合力受体(p75NTR)、α1-糖蛋白、氧调节蛋白1、钙蛋白酶抑制剂、假想蛋白-10表达上调、下调与消失变化在脊髓损伤中居极其重要地位。更多还原

【Abstract】 Objective To establish the two-dimensional gel electrophoresis(2-DE) and Computer Image analysis is for the proteome research of acute spinal injury,and to compare and observe the its proteome expression with the acute spinal injury.Methods To establish animal model of the spinal injury,and its total protein were analysed and extracted by 2-DE,sliver staining,Image Master 2D 5,0 analysis software,matrix assisted laser desorption ionization-time of light mass spectrometry(MALDI-TOF-TOF-MS)and were used to separate and identification the proteome from protein in 2DE.Results We obtained proteins of the satisfactory 2DE patterns of the spinal cord.12 protein spots were mass chromatographic analysis.and 9 protein spots were identified by MALDI-TOF-TOF-MS.These up-regulation proteins in the spinal cord injury group present increase tendency,however,calpai,neurofilament proteinM,neurenergen lower-avidity acceptor(p75NTR),α1-Acid-glycoprotein and oxygen regulated protein 1 appeared decrease.tendency,and calpain inhibitor Hypothetical protein cgi-10 in the spinal cord injury group were disappear.Conclusion We have identified 7 kind of protein during the spinal cord inury,(include calpain,neurofilament protein,neurenergen lower-avidity acceptor α1-Acid-glycoprotein and oxygen regulated protein 1,and calpain inhibitor Hypothetical protein cgi-10 etc),their changed of the up-regulation,down regulation,and to disappear were play important roly in the spinal cord injury.更多还原