冠脉搭桥手术心脏停跳后心肌缺血损伤蛋白质组学研究.DOC

人低温离体灌注心脏心肌缺血损伤早期蛋白组学研究 李继元1,2 李洪1 杨天德1 (400037 重庆，第三军医大学新桥医院麻醉科1；024053 内蒙古 赤峰，内蒙古赤峰市65657部队卫生队2) [摘要] 目的：分析离体心脏灌注早期心肌缺血损伤释放出的蛋白质组成，并探讨这些蛋白的生理学过程和功能，为进一步寻找心肌缺血损伤早期标志物奠定基础。方法：收集人离体心脏停跳后即刻、30min和60min后的4℃灌注液，样品首先用MARS柱去除高丰度蛋白，用10 KD分子筛脱盐浓缩、溶液内酶解，得到肽段用mRP柱进一步脱盐，通过液相-芯片-离子阱质谱（HPLC-Chip-MS/MS）技术进行鉴定和生物信息学分析，建立离体心脏灌注早期心肌缺血释放蛋白全谱图，通过GO注释和EBI数据库对这些蛋白进行功能和定位分析以及BLAST比对。结果：利用Agilent Spectrum Mill 蛋白数据库专业统计分析处理软件进一步研究HPLC-Chip-MS/MS；生物信息学分析 Study of proteomics in early ischemic myocardium of isolated human heart with hypothermia-perfusion Li Jiyuan1,2, Li Hong1, Yang Tiande1．（1Department of Anesthesiology, Xinqiao Hospital ,Third Military Medical University, Chongqing ,400037；2Medical Team of PLA No. 65657 Unit ,Inner Mongolia Chifeng ,024053,China） [Abstract] Objective To analyze the component of proteins released from early ischemic myocardium of isolated human heart with hypothermia perfusion，and to explore the physiological process and function of these proteins by GO annotation for further searching of early markers of myocardial ischemia injury. Methods Perfusates from human hearts were harvested instantly, at 30 min and 60 min after cardioplegic solution perfusion. To remove high abundance proteins from samples with MARS columns, and to desalinize with 10 KD molecular sieve to digested in solution, and the obtained peptide fragments were desalinized by mRP columns, then all peptide fragments were identified by HPLC-Chip-MS/MS and Bioinformatics was used to analyze the identified Proteins. All released Proteins of early myocardial injury were Picked out and analyzed by GO and EBI analysis. Results we obtained 176 high confidence proteins (Score ≧8, SPI ≧ 70%) by Using Agilent Spectrum Mill Home software , after GO annotation , these proteins mainly participated in cell process, biological regulation, metabolism and other physiological processes, with blinding function, catalytic activity and activity of enzyme regulation; through the EBI database and we foun