Ⅱ型糖尿病大鼠神经视网膜组织中差异蛋白质分析  

Differential Proteomic Analysis of Neural Retina Proteins in Type 2 Diabetes Rats

作　　者：王亚冬[1] 童明庆[1] 崔毓贵[1] 张民英[2] 郭雪江[2] 王玲[2] 许志洋[2] 王富强[2] 

机构地区：[1]南京医科大学第一附属医院,南京210029 [2]南京医科大学基础医学院,南京210029

出　　处：《分析化学》2010年第10期1462-1468,共7页Chinese Journal of Analytical Chemistry

基　　金：江苏省教育厅自然科学基金(No.08KJB320005)资助

摘　　要：以高脂饮食联合小剂量链脲菌素(Streptozotocin,STZ)注射方法诱导大鼠II型糖尿病(T2DM)模型为研究对象,采用比较蛋白质组学技术鉴定和分析早期糖尿病神经视网膜差异表达蛋白。通过联用液氮内研磨、冰浴超声、高速离心提取全蛋白技术,提取每只大鼠神经视网膜全蛋白质总量约900μg。采用双向凝胶电泳(2-DE)分离技术,获得2500个以上可辩认的蛋白质斑点。通过蛋白质组学比对技术筛选出糖尿病状态下神经视网膜差异表达蛋白,用基质辅助激光解吸电离飞行时间串联质谱(MALDI-TOF-MS/MS)和肽质量指纹图谱(PMF)鉴定出20个差异蛋白点。按照Gene Ontology(GO)分类体系,对差异蛋白归类分析,揭示其亚细胞定位和分子功能。应用Pathway Studio软件对差异表达蛋白的生物学意义进行分析,认为凋亡是早期糖尿病视网膜病变(Diabetic retinopathy,DR)重要的细胞事件;AnnexinΙ、CRYAB、mtHsp70蛋白是与病理过程相关的关键蛋白,对研究DR致病机制有重要意义。The rats of model type 2 diabetes mellitus （T2DM） induced by high-fat diet and small dose injection of streptozotocin （STZ） were taken as the research subject.A comparative proteomics approach was adopted to identify and analyze proteins differentially expressed in early diabetic neural retina.The total retinal protein extracts were obtained by grinding tissue samples in liquid nitrogen combined with ice bath ultrasonic and ultra-centrifugation.The amount of protein extracted from retinas of each rat was approximately 900 μg, and more than 2500 protein spots were observed in the following two-dimensional gel electrophoresis （2-DE） analysis.The comparative proteomic data revealed differentially expressed protein spots in the diabetic neural retina.Of which,twenty differentially expressed protein spots were successfully identified by tandem mass spectrometry （MS/MS） as well as by peptide mass fingerprinting （PMF） .The identified proteins were then categorized based on their sub-cellular localizations and biological functions using Gene Ontology （GO） .Further analysis using Pathway Studio （Ariadne Genomics） indicated that apoptosis was a predominant cellular response in the pathological process of diabetic retinopathy （DR） .Key proteins involved in this pathological process,including Annexin Ι,CRYAB,and mtHsp70,were identified in this study.

关 键 词：大鼠神经视网膜 比较蛋白质组学 双向凝胶电泳 基质辅助激光解析电离飞行时间质谱 糖尿病 

分 类 号：R587.2[医药卫生—内分泌] R774.1[医药卫生—内科学]