大鼠烫伤模型肠黏膜蛋白质组的分离鉴定及其功能分析  

Separation,identification and functional analysis of the proteomics of intestinal mucosa in scalded rats

作　　者：王晓军[1] 孙永华[2] 王建宁[1] 宋国成[1] 高继东[1] 殷岳[1] 郑明非[1] 

机构地区：[1]北京市第六医院外二科,100007 [2]北京市积水潭医院

出　　处：《中华损伤与修复杂志（电子版）》2007年第4期219-224,共6页Chinese Journal of Injury Repair and Wound Healing(Electronic Edition)

基　　金：国家重点基础研究发展规划（973）项目（No：G19990542）

摘　　要：目的研究严重烧伤后早期肠黏膜蛋白质组的整体变化特性及功能,并对明确的蛋白质进行临床意义分析,为探讨严重烧伤后肠黏膜损害的发病机制和促进肠黏膜的修复提供理论依据。方法采用大鼠Ⅲ度烫伤模型,利用高分辨双向电泳(2-DE)对肠黏膜组织进行蛋白质分离,Im- ageMaster 2D Elite图像分析软件进行分析,应用生物质谱、蛋白质库及文献的分析等技术研究大鼠烫伤后肠黏膜蛋白质组的变化特性。结果(1)烧伤后6h和12h两组明确下调的蛋白质点数为34,进行鉴定及分析的蛋白质22个,分别参与线粒体的变化(如线粒体乌头酸酶、丙酰辅酶A羧化酶、肝脏F1ATP酶重链A、短链羟酰基辅酶A脱氢酶、P-电子转移[传递]黄素蛋白α亚基六种)、参与代谢(如磷酸丙糖异构酶1和细胞溶质环氧化物水解酶)、细胞骨架(如纤维单元素、类动力蛋白-5、肌钙蛋白-2和碱性肌浆球蛋白轻链3四种)、参与调控(如糖皮质激素诱导蛋白、核因子1-B2、BRCA1、雌二醇安息香酸盐转录因子、G-蛋白β-2亚基、N-甲基-D-天门冬氨酸受体1六种)和免疫调控(如T细胞受体-V-δ6和Ig重链V区蛋白1);(2)烧伤后6 h和12 h两组明确上调的蛋白质点数为12,进行检测及功能分析的蛋白质10个。参与了应激反应(如糖调控蛋白前体78、蛋白质二硫化物异构酶A3 (PDIA3)前体、基质金属蛋白酶11、α-烯醇酶和鸟氨酸转化酶)和其他蛋白质(如白蛋白、乳酸脱氢酶A、T细胞活化连接蛋白、辅酶Q和eps8捆绑蛋白)的变化。结论本研究从和线粒体有关的蛋白质的改变、骨架及基质蛋白的变化、代谢的调控、激素因子分泌的信号调控、免疫反应、应激反应和其他蛋白质的改变等7个方面揭示了严重烧伤后早期肠黏膜屏障的病理生理变化特性,以线粒体和对激素及因子的调控的病理生理改变最为突出。Objectives To explore the function and the features of the integral changes in the proteomics of intestinal mucosa after severe burn injury, and to analyze the clinical significance of the clarified proteins, so as to investigate the pathogenesis of the intestinal mucosal injury after severe burn injury and to provide theoretical basis for the repair of intestinal mucosa. Methods The rats inflicted by 30% TBSA of full-thickness scalding were employed as the model. The high resolution two-dimensional electrophoresis （2- DE） was employed to separate the proteins in the intestinal mucosal tissue. And the proteins were analyzed by ImageMaster 2D Elite image analysis software. The features of the changes of the proteomics of rat intestinal mucosa after scalding were studied by biological spectrometry, protein bank and reference article analysis technique. Results The down-regulated proteomics points were 34 in both groups at 6 and 12 postburn hours （PBHs）. Twenty-two kinds of proteins were identified and analyzed, which respectively took part in the change of mitochondria （ such as mitochondrial aconitase,alpha-propionyl -CoA carboxylase, heavy chain A, rat liver F1-Atpase ,troponin-1 ,hydroxylacyl-coenzyme A dehydrogenase, short chain and alpha-subunit of P- electron transfer flavoprotein（ETF） ） . Some participated in the metabolism （ such as triosephosphate isomerase 1 and cytosolic epoxide hydrolase）. Some took part in the formation of cytoskeleton （ such as fibromodulin, dynein-like protein 5 ,Troponin-2, myosin light chain 3 alkali（ MLC ） ） . Some participated in the modulation （such as glucocorticoid-inducible protein , NF1-B2, BRCA1, transcription factor EB （estradiol benzoate ） ,G-protein beta-2 subunit, N-methyl-D-aspartate receptor 1 （NMDAR））and some joined the immunological modulation （ such as T cell receptor Ⅴ delta 6 and Ig heavy chain V region （ V1 ） ） . The evident up-regulated proteomics points in both groups were 12 during 6 and 12 PBHs. There were 10

关 键 词：烧伤 肠黏膜 蛋白质组 双向凝胶电泳 

分 类 号：R644[医药卫生—外科学]