Department of Ophthalmology, Shanghai tenth People's Hospital, Tongji University School of Medicine, 301 Yanchang Road, Shanghai, 200072, China.
Tongji University School of Medicine, 1239 Siping Road, Shanghai, 200092, China.
Proteome Sci. 2012 Mar 5;10(1):15. doi: 10.1186/1477-5956-10-15.
Diabetes can lead to serious microvascular complications such as proliferative diabetic retinopathy (PDR), which results in severe vision loss. The diabetes-induced alterations in the vitreous protein composition in diabetic patients with PDR may be responsible for the presence of PDR. The vitreous humour can be utilised in a variety of studies aimed toward the discovery of new targets for the treatment or prevention of PDR and the identification of novel disease mechanisms. The aim of this study was to compare the protein profile of vitreous humour from diabetic patients with PDR with that of vitreous humour from normal human eyes donated for corneal transplant.
Vitreous humour from type 2 diabetic patients with PDR (n = 10) and from normal human eyes donated for corneal transplant (n = 10) were studied. The comparative proteomic analysis was performed using two-dimensional fluorescence difference gel electrophoresis (2-D DIGE). Differentially produced proteins (abundance ratio > 2 or < -2, p < 0.01) were identified by matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI-TOF MS) and MALDI-TOF tandem mass spectrometry. A total of 1242 protein spots were detected on the 2-D master gel of the samples, and 57 spots that exhibited statistically significant variations were successfully identified. The spots corresponded to peptide fragments of 29 proteins, including 8 proteins that increased and 21 proteins that decreased in PDR. Excluding the serum proteins from minor vitreous haemorrhage, 19 proteins were found to be differentially produced in PDR patients compared with normal subjects; 6 of these proteins have never been reported to be differentially expressed in PDR vitreous: N(G),N(G)-dimethylarginine dimethylaminohydrolase 1 (DDAH 1), tubulin alpha-1B chain, gamma-enolase, cytosolic acyl coenzyme A thioester hydrolase, malate dehydrogenase and phosphatidylethanolamine-binding protein 1 (PEBP 1). The differential production of pigment epithelium-derived factor (PEDF) and clusterin was confirmed by Western blot analysis.
These data provide an in-depth analysis of the human vitreous proteome and reveal protein alterations that are possibly involved in the pathogenesis of PDR. Further investigation of these special proteins may provide potential new targets for the treatment or the prevention of PDR.
糖尿病可导致严重的微血管并发症，如增生性糖尿病视网膜病变（PDR），从而导致严重的视力丧失。糖尿病患者 PDR 玻璃体蛋白组成的改变可能是 PDR 存在的原因。玻璃体可以用于各种旨在发现 PDR 治疗或预防新靶点以及鉴定新疾病机制的研究。本研究的目的是比较 PDR 糖尿病患者的玻璃体蛋白谱与正常供体角膜移植的人眼玻璃体蛋白谱。
研究了 10 例 2 型糖尿病 PDR 患者和 10 例正常供体角膜移植的玻璃体。采用二维荧光差异凝胶电泳（2-D DIGE）进行比较蛋白质组学分析。通过基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）和 MALDI-TOF 串联质谱鉴定差异表达的蛋白质（丰度比> 2 或< -2，p < 0.01）。在样本的 2-D 主凝胶上共检测到 1242 个蛋白点，成功鉴定出 57 个具有统计学意义的差异表达点。这些斑点对应于 29 个蛋白质的肽片段，其中包括在 PDR 中增加的 8 种蛋白质和减少的 21 种蛋白质。排除轻微玻璃体出血的血清蛋白后，与正常对照组相比，PDR 患者有 19 种蛋白质差异表达；其中 6 种蛋白质在 PDR 玻璃体中从未报道过差异表达：N(G)，N(G)-二甲基精氨酸二甲氨基水解酶 1（DDAH 1）、微管蛋白 alpha-1B 链、γ-烯醇酶、胞质酰基辅酶 A 硫酯水解酶、苹果酸脱氢酶和磷酸乙醇胺结合蛋白 1（PEBP 1）。PEDF 和载脂蛋白 E 的差异产生通过 Western blot 分析得到证实。
这些数据提供了对人玻璃体蛋白质组的深入分析，并揭示了可能参与 PDR 发病机制的蛋白质改变。对这些特殊蛋白质的进一步研究可能为 PDR 的治疗或预防提供潜在的新靶点。
