大鼠急性脊髓牵张损伤相关蛋白质的筛选和鉴定  

Screen and identification of the relative protein expressed in the acute tractive spinal cord injury in rats

作　　者：王文岳[1] 杨天府[1] 刘雷[1] 雷鸣鸣[1] 

机构地区：[1]四川大学华西医院骨科,成都610041

出　　处：《中华外科杂志》2012年第8期737-743,共7页Chinese Journal of Surgery

基　　金：国家自然科学基金资助项目（0040205401265）

摘　　要：目的筛选并鉴定与大鼠急性脊髓牵张损伤相关的蛋白质。方法sD大鼠10只随机分为2组：假手术组（Sham组）和脊髓牵张损伤组（tractivespinalcordinjury，TSCI组），每组5只。术后1d取材，对脊髓组织蛋白进行提取制备、定量、分离，建立差异表达图谱，通过双向凝胶成像分析和人工比对，分别确定差异蛋白质斑点。采用纳升超高效液相色谱-电喷雾串联质谱技术鉴定差异蛋白，检索数据库，对差异蛋白进行分类。结果Sham组与TSCI组两者间完全相同的差异蛋白质点共22个，其中4个下调，18个上调；4个差异蛋白点新出现在TSCI组。鉴定出16种有意义的蛋白质，其中与凋亡相关的蛋白4种，神经信号转导相关的蛋白3种，参与代谢的蛋白6种，未知蛋白3种。结论鉴定出的蛋白质可能通过神经信号转导、调节细胞凋亡及细胞代谢等途径参与了TSCI的损伤及修复过程。Objective To screen and identify the relative protein expressed in the acute tractive spinal cord injury （TSCI） in rats. Methods Ten adult Sprague Dawley rats were randomly divided into Sham group and TSCI group, 5 rats in each group. Rats from Sham group and TSCI group at 1 day after surgery were sacrificed for harvesting T13-L2 spinal tissue specimens. The extraction and quantitation of protein in the spinal tissue was finished firstly. Proteins from spinal tissue were separated by two-dimensional eleetrophoresis （2-DE） and identified by mass spectrometry （MS）. The different expression map was established in each group, and proteins express differently was determined by comparing the level of each spot with gel imaging software and manually. Proteins were identified by High performance liquid chromatography-electrospray tandem （NanoUPLC-ESI-MS/MS） and peptide sequence tag with tandem MS combining with database respectively. After that, the function of these identified proteins was known and classified. Results There were 22 differential protein expression spots were found between Sham group and TSCI group. Among them, 18 spots were up-regulated and 4 were down-regulated. 4 differential protein expression spots were newly found in TSCI group. Sixteen significant proteins were identified by NanoUPLC- ESI-MS/MS. Four kind of proteins were related to apoptosis, 3 in nerve signal transduction and 6 in metabolism, respectively. Unnamed proteins were 3. Conclusions The differential expression proteins were found between Sham group and TSCI group. These identified proteins may play important role in the process of injury and recovery through transducting nerve signal, regulating nerve cells apoptosis and metabolism.

关 键 词：脊髓损伤 蛋白质组学 色谱法 高压液相 光谱法 质量 电喷雾电离 大鼠 

分 类 号：R651.2[医药卫生—外科学]