食源性肥胖大鼠下丘脑差异表达蛋白的蛋白质组学分析  

Proteomic Analysis of Differentially Expressed Proteins of Hypothalamus in Diet-induced Obesity's Rat

作　　者：王启明[1] 田德润[1] 魏中南[1] 蔡滢[1] 王飞[2] 韩济生[2] 

机构地区：[1]天津医科大学解剖学教研室,天津300070 [2]北京大学神经科学研究所,北京100191

出　　处：《中国生物化学与分子生物学报》2009年第8期740-746,共7页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金项目(No.3087791);天津市应用基础研究计划项目(No.07JCZD08100)~~

摘　　要：建立食源性肥胖大鼠模型,对正常大鼠和肥胖大鼠下丘脑全蛋白进行双向凝胶电泳,产生下丘脑蛋白双向凝胶电泳图谱.对图谱进行比对分析后,从凝胶上切取差异表达的蛋白点,经胶内酶解,通过基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)对酶解后的肽段进行分析,再经数据库(NCBInr)检索,对蛋白质进行鉴定.研究发现,正常组表达图谱可检测到1160±15(n=5)个蛋白点,肥胖组表达图谱可检测到1070±10(n=5)个蛋白点,与对照组相比,匹配率大于80%.并且成功鉴定了17种差异表达蛋白质,其中有7种在肥胖组表达上调,10种表达下调.它们分别属于代谢酶、细胞周期调控因子、抗氧化蛋白、信号传导蛋白、蛋白酶体相关蛋白、细胞骨架蛋白以及未知蛋白等.与正常对照组相比,肥胖组的蛋白质表达存在着较大差异,通过对差异表达蛋白的分析,提示了在肥胖发生的过程中,下丘脑神经中枢经历了一个非常复杂的信号活动和特定改变,为深入认识肥胖的发病机制奠定了基础.One model of diet-induced obesity （DIO） rat had been built initially. Proteins from hypothalamus of the DIO and normal rats were separated by two-dimensional gel electrophoresis （2-DE）, respectively. The proteins were stained with Coomassie brilliant blue to produce a high-resolution map of the rat hypothalamus proteome. The differentially expressed proteins from this map were digested with trypsin, then were analyzed by matrix-assisted laser desorption / ionization time of flight mass spectrometry （MALDI-TOF-MS）. The mass spectrometric data were used to identify the proteins through searches on line of the NCBInr protein sequence database. Through the analyses, 1 160 ± 15 （n = 5） protein spots were distinguishable in proteome map of normal rats, 1 070 ± 10 （ n = 5 ） protein spots were distinguishable in proteome map of DIO rats. The matching rate of protein spots in DIO rats were more than 80% compared with that of normal rats. By the analyses of match differentially displayed map, 17 prominent （7 up-regulated and 10 down-regulated） proteins, each being unique in DIO rats were identified. These identified proteins were classified into metabolism enzymes, modulins of cell cycle, cytoskeleton proteins, antioxidant proteins, signaling proteins, proteasome related proteins, specific proteins of neurocyte and unknown proteins. This study provides characterization of proteins of the rat hvpothalamus and will be used for future studies of differential protein expression in DIO rat models. The data would help illuminate the mechanisms involved in the progress of obesity in hypothalamus nervous centre.

关 键 词：下丘脑 食源性肥胖 蛋白质组学 双向凝胶电泳 质谱 

分 类 号：R322[医药卫生—人体解剖和组织胚胎学]