Proteomic analysis of plasma proteins in diabetic retinopathy patients by two dimensional electrophoresis and MALDI-Tof-MS
Proteomic analysis of plasma proteins in diabetic retinopathy patients by two dimensional electrophoresis and MALDI-Tof-MS
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DOI：
10.1016/j.jdiacomp.2015.05.021
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发表时间：
2015-09-01
影响因子：
3
通讯作者：
Bhaskar, Anusha
中科院分区：
文献类型：
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作者：
Gopalakrishnan, Vidhya;Purushothaman, Parthiban;Bhaskar, Anusha
目的：糖尿病视网膜病变是糖尿病的一种高度特异的血管并发症，从以血管通透性增加为特征的轻度非增殖性异常发展到以视网膜血管生长为特征的中、重度增殖性糖尿病视网膜病变。本研究的目的是用双向凝胶电泳法鉴定糖尿病视网膜病变患者的差异表达蛋白质。方法：采集糖尿病患者(无视网膜病变)和糖尿病视网膜病变患者的血样，以EDTA为抗凝剂。白蛋白和免疫球蛋白免疫球蛋白被共同去除，以富含丰度较低的蛋白质。结果：正常对照组和糖尿病视网膜病变组分别有48个和61个蛋白质点表达下调，其中视黄醇结合蛋白1(retinol-bindingProtein 1，RBP1)、肌醇多聚磷酸酶3α(NUD10)、脑红蛋白(NGB)表达下调，而血红蛋白(Hb)和CD160抗原(CD160抗原)表达上调。切下这五个蛋白点，进行凝胶内胰酶消化，并用UltraFlex MALDI-TOF-MS确定它们的身份。结论：我们报道了一种基于患者的综合血浆蛋白质组学方法，用于糖尿病视网膜病变的筛查和检测。研究的意义：我们确定了5种不同的蛋白质，这些蛋白质在对照组糖尿病患者(无视网膜病变)的血浆中差异表达。在这5种蛋白中，脑红蛋白(NGB)蛋白的表达差异显着，可能成为糖尿病视网膜病变的潜在生物标志物。(C)2015 Elsevier Inc.保留所有权利。
Objective: Diabetic retinopathy is a highly specific vascular complication of diabetes mellitus and progresses from mild non-proliferative abnormalities characterized by increased vascular permeability to moderate and severe proliferative diabetic retinopathy characterized by the growth of blood vessels on the retina. The aim of the study was to identify the differentially expressed proteins in diabetic retinopathy using two-dimensional electrophoresis.Methods: Blood sample was drawn from subjects with diabetes mellitus (without retinopathy) who served as controls and patients with diabetic retinopathy in tubes containing EDTA as anticoagulant. Albumin and immunoglobulin IgG collectively removed to enrich proteins of lower abundance. 2de was carried out to see if there are any differentially expressed proteins.Results: Approximately 48 and 61 spots were identified in control and diabetic retinopathy respectively, of which three protein spots RBP1 (retinol-binding protein 1), NUD10 (Diphosphoinositol polyphosphohy-drolase 3 alpha), NGB (neuroglobin) were down regulated and HBG2 (hemoglobin) and BY55 (CD 160 antigen) were upregulated in diabetic retinopathy. These five protein spots were excised and were subjected to in-gel tryptic digestion, and their identities were determined by ultraflex MALDI-TOF-MS.Conclusion: We report a comprehensive patient-based plasma proteomic approach to the identification of potential biomarkers for diabetic retinopathy screening and detection. Significance of the study: We identified 5 different proteins that were differentially expressed in the plasma of control diabetic patients (without retinopathy). Among these five proteins the expression of neuroglobin (NGB) protein varied significantly and may be a potential biomarker in diabetic retinopathy. (C) 2015 Elsevier Inc. All rights reserved.