【作者】 李琦；

【导师】 张学东；

【作者基本信息】 重庆医科大学 ， 眼科学， 2008， 硕士

【摘要】 目的对增生期糖尿病视网膜病变(Proliferative diabetic retinopathy,PDR)伴重度黄斑水肿的玻璃体蛋白质组学变化进行初步研究。方法在睫状体平坦部穿刺抽取27例PDR伴重度黄斑水肿及9例健康人的玻璃体液,用双向凝胶电泳( 2-DE)的方法得到2-DE图谱,PD Quest7.3.1软件对比图谱,获得差异蛋白点。将差异蛋白点取出、酶解后,用表面增强激光解吸离子化-飞行时间质谱(SELDI-TOF-MS)的方法对玻璃体差异蛋白质进行初步鉴定,得到肽质量指纹图(PMF),采用Mascot和MS-Fit软件进行数据检索,找到差异蛋白;用Western blot行差异蛋白质的蛋白质印迹验证。结果PDR伴重度黄斑水肿的玻璃体和正常人玻璃体的蛋白质图谱PDQuest软件比较,蛋白点强度相差2倍以上且有统计学意义(P<0.01)的共有30个,其中20个蛋白点上调,10个蛋白点下调。选取上调最明显的10个蛋白点进行质谱分析和数据库的检索及鉴定,其中4个明显差异的蛋白质即11-视黄醇结合蛋白、18-载脂蛋白D、20-载脂蛋白AIV、28-甲状腺激素结合蛋白。结论初步建立PDR伴重度黄斑水肿的玻璃体的蛋白质2-DE图谱。其蛋白质组发生了改变,本研究发现有四种蛋白质明显增加。更多还原

【Abstract】 PURPOSE: To identify and analyze proliferative diabetic retinopathy with heavy macular edema-related proteins in the vitreous.METHODS: The vitreous samples of 27 eyes active proliferative diabetic retinopathy with heavy macular edema patients with 9 normal eyes vitreous samples were analyzed by two-dimensional gel electrophoresis then compared the 2-DE map with PDquest to find the disparation proteins spots. The spots were extracted and been enzymolysis and progressing mass spectrogram identification. Then according the results of Peptide Mass Finger printing(PMF) to find the disparation proteins by searching internet using Mascot and Ms-Fit. One sample of the each group was enrolled to identify disparation proteins by Western blot.RESULTS: At least 30 proteins were up regulated or down- regulated by over two folds between two groups. 20 protein spots up regulated and 10 down- regulated .Four proteins (apolipoprotein A-IV precursor, Transthyretin precursor, apolipoprotein D , Retinol-binding protein I, cellular) in the 20 spots were expressed significantly differently between the normal and PDR with heavy macular edema patients. The four proteins were up regulated in the diabetic retinopathy with macular edema patients.CONCLUSIONS: We constructed vitreous protein profiles for proliferative diabetic retinopathy with heavy macular edema patients and identified four candidate proteins believed to be involved in the pathogenesis of proliferative diabetic retinopathy with heavy macular edema.更多还原