糖尿病性白内障大鼠晶状体蛋白质的质谱分析

发布时间：2018-03-13 20:12

本文选题：糖尿病性白内障　切入点：晶状体蛋白　出处：《吉林大学》2012年硕士论文　论文类型：学位论文

【摘要】：糖尿病性白内障是糖尿病患者眼部最常见的并发症之一，也是糖尿病患者致盲的重要原因。与年龄相关性白内障相比，糖尿病患者发病较早，且进展迅速。在基因水平上对白内障形成因素的研究较多,而有关蛋白质组学的研究则很少见,但是只局限于基因水平来研究白内障的形成是远远不够的,通过研究由基因编码和翻译的蛋白质,才可能真正揭示白内障的发病机制。后基因组时代,蛋白质组学作为功能基因组学研究的核心技术之一,已成为生物医学领域强大的技术支撑,并为白内障疾病的研究提供了一个新的平台。目的：本实验研究采用蛋白质组学方法对糖尿病性白内障晶状体核不同组分晶状体蛋白的组成进行研究，探讨糖尿病性白内障的发病机制。方法：运用小剂量链脲佐菌素（STZ）+特殊膳食喂养法饲养同一周龄的ＳD大鼠20只，，每组10只，另取10只同周龄SD大鼠作对照组。后分别取造模后6周、20周及对照组大鼠晶状体各10只作为样本。提取晶状体核中的水溶性蛋白，应用蛋白质组学手段，即荧光差异凝胶电泳（2D-DIDG）、图像分析、MALDI TOF MS分析及数据库搜索，进行差异表达蛋白质的研究。结果：荧光差异凝胶电泳结果显示,糖尿病性白内障晶状体核中水溶性蛋白大部分为相对分子质量14400~97400, pI3~9。高丰度蛋白质斑点相对分子质量分布在20000~31000, pI4~7区域内。糖尿病性白内障晶状体蛋白二维电泳图识别出65个差异点，其中上调表达39个，下调表达26个。其中差异明显的点（差异量达3倍以上）共19个。选取重复性及分辨率较好的图谱进行后续的图谱分析。鉴定出5种蛋白质,其中2种蛋白质表达上调(β-A3晶状体蛋白、β-A4晶状体蛋白)，3种蛋白质表达下调(β-B2晶状体蛋白、α-A2晶状体蛋白、α-B2晶状体蛋白)。结论：本研究获得了糖尿病性白内障大鼠蛋白质差异凝胶电泳图谱，并成功鉴定了部分蛋白斑点,其中2种蛋白质表达上调(β-A3晶状体蛋白、β-A4晶状体蛋白)，3种蛋白质表达下调(β-B2晶状体蛋白、α-A晶状体蛋白、α-B晶状体蛋白)，研究有关糖尿病性白内障形成过程中蛋白质表达的差异，从新的角度探讨糖尿病性白内障形成的病理机制,以期为白内障的预防和药物治疗提供更多的依据和途径。
[Abstract]:Diabetic cataract is one of the most common complications in the eyes of diabetic patients, and it is also an important cause of blindness in diabetic patients. There are many studies on the factors of cataract formation at the gene level, while proteomics studies are rare, but it is far from enough to study cataract formation only at the gene level. By studying proteins encoded and translated by genes, it is possible to truly reveal the pathogenesis of cataract. In the post-genome era, proteomics is one of the core technologies in functional genomics. Has become a powerful technical support in the field of biomedicine, and provides a new platform for the research of cataract diseases. In this study, proteomics was used to study the composition of lens nuclei of diabetic cataract, and to explore the pathogenesis of diabetic cataract. Twenty Sprague-Dawley rats of the same week age were fed with a special diet with low dose streptozotocin (STZ) for 10 rats in each group. In addition, 10 SD rats of the same age were taken as control group. The lens samples were taken from the rats 6 weeks to 20 weeks after model making and 10 rats from the control group respectively. The water-soluble protein in the lens nucleus was extracted, and the proteomics method was used to extract the water-soluble protein from the lens nucleus. The results showed that: fluorescence differential gel electrophoresis 2D-DIDGG, image analysis, MALDI TOF MS analysis and database search were used to study the differential expression of proteins. The results of fluorescence differential gel electrophoresis showed that, Most of the water-soluble proteins in the lens nucleus of diabetic cataract were relative molecular weight 14400 ~ 97400 and Pi _ 3N _ 9. The distribution of relative molecular weight of high abundance protein spots was in the region of 20000n31000, pI4~7. Two-dimensional Electrophoretic Mapping of Diabetic Cataract Crystalloprotein. Don't make 65 differences, Among them, 39 were up-regulated and 26 were down-regulated. Among them, there were 19 points with obvious difference (the amount of difference was more than 3 times). Two proteins were up-regulated (尾 -A3, 尾 -A4, 尾 -A4) and three proteins were down-regulated (尾 -B2, 伪 -A2, 伪 -B2, 伪 -B2, 伪 -B2). In this study, the protein differential gel electrophoresis patterns of diabetic cataract rats were obtained, and some protein spots were successfully identified. Two proteins were up-regulated (尾 -A3, 尾 -A4, 尾 -A4, 尾 -A4, 尾 -A4, 尾 -B2, 伪 -A, 伪 -B, 伪 -B, etc.) in order to study the difference of protein expression in diabetic cataract formation, and the expression of 尾 -A3, 尾 -A4 and 尾 -A4 were down-regulated (尾 -B2, 伪 -A, 伪 -B, 伪 -B). To explore the pathological mechanism of diabetic cataract from a new point of view, in order to provide more evidences and approaches for the prevention and drug treatment of cataract.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R776.1;R587.1

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