摘要
目的建立急性脊髓损伤双向电泳图,分析损伤后脊髓内蛋白质的差异表达。方法使用裂解液提取脊髓蛋白,以双向电泳技术分离蛋白质,凝胶染色、图象分析后选取蛋白点,通过质谱技术获取肽质量指纹谱,在蛋白质数据库中查询蛋白质。结果成功建立重复性较好的脊髓蛋白质组双向电泳图,找出13个表达明显变化的蛋白点并获得这些蛋白质的肽质量指纹谱,成功鉴定出11个蛋白质。结论成功构建大鼠正常脊髓与损伤脊髓双向电泳图,用质谱技术鉴定了部分蛋白质。这些蛋白质的有些功能最近才被人们认识。相信在脊髓损伤后的继发性损伤中这些蛋白质可能会起到重要的作用。
Objective To establish two dimensional eleetrophoresis （2DE） gels of acute spinal cord injury （SCI） of rats and analyze differential expressions of proteins in spinal cord. Methods Protein extraction of spinal cord was gained by lysis buffer and 2DE technology was adopted. After Coomassie Brilliant Blue B-250 staining and ImageMaster software analysis, matrix assisted laser desorption ionization-time of flight-mass spectrometry （MALDI-TOF-MS） and database search were used to identify the interest protein spots in 2DE gels. Results 2DE gels of acute SCI with good reproducibility have been suecessfuUy established. 13 proteins were found to express differentially and their poptide mass fingerprints （PMFs） were also obtained. 11 proteins were successfully identified. 10 proteins were up-regulated, while 1 protein was down-regulated. Conclusion 2DE gels of normal spinal cord and injured spinal cord have been successfully established. Some new functions of the identified proteins have just been recognized. These proteins may play important part in the secondary injury after SCI .
出处
《脊柱外科杂志》
2008年第2期99-103,共5页
Journal of Spinal Surgery
基金
国家自然科学基金资助项目(30572165)
关键词
大鼠
脊髓损伤
电泳
凝胶
双向
光谱分析
质量
Rats
Spinal cord injuries
Electrophoresis, gel, two-dimensional
Spectrum analysis, mass
