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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.
Establishment of food-induced obesity rat model, the normal rat and obese rat hypothalamic protein by two-dimensional gel electrophoresis, hypothalamic protein two-dimensional gel electrophoresis pattern generated after mapping analysis, from the gel cut The differentially expressed protein spots were digested by gel and analyzed by matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The peptides were digested by NCBInr, The results showed that 1160 ± 15 (n = 5) protein spots were detected in the normal group and 1070 ± 10 (n = 5) spots in the obesity group, compared with the control group Rate of more than 80%, and successfully identified 17 kinds of differentially expressed proteins, of which 7 were up-regulated in obesity group, 10 were down-regulated.These belong to metabolic enzymes, cell cycle regulators, antioxidant proteins, signal transduction proteins, proteases Body-related proteins, cytoskeletal proteins and unknown proteins, etc .. Compared with the normal control group, the protein expression of obese group there is a big difference, through the analysis of differentially expressed proteins, suggesting that in the process of obesity, hypothalamic nerve The center experienced a very complex signal activity and specific changes, which laid the foundation for understanding the pathogenesis of obesity.