Proteomic analysis of vitreous from diabetic macular edema and development of its therapy.

目的：应用双向凝胶电泳和质谱技术对糖尿病性黄斑水肿（DME）患者玻璃体中的相关蛋白进行鉴定和分析。16例DME患者的黄斑前区采集玻璃体标本0.5ml。DME患者玻璃体中的蛋白质浓度显著高于非DME患者（1436 ± 337 vs. 967 ± 217 μ g/ml，p = 0.05）。对15 μ g样品进行2D凝胶电泳。用SYPRO-Ruby蛋白质染色后，使用图像分析软件确定斑点强度。从凝胶上切下清晰可见的斑点，用胰蛋白酶原位消化，并通过液相色谱串联MS（LC MS/MS）序列分析鉴定蛋白质。我们确定了8个点，显着高于非DME样品的DME。从8个点中，鉴定出6种蛋白质，即PEDF、ApoA-4、ApoA-1、Trip-11、PRBP和维生素D结合蛋白。其中一个斑点仅在非DME中表达，并被鉴定为Apo-H。我们认为，这些化学介质存在于后玻璃体中可能在DME的发病机制中发挥作用。

To identify and analyze diabetic macular edema (DME)-related proteins in the vitreous, en masse, using two-dimensional gel (2D gel) electrophoresis and mass-spectrometry (MS). Vitreous samples (0.5ml) were collected from the pre-macular portion of 16 DME cases. The protein concentration in vitreous from patients with DME was significantly higher than in non-DME patients (1436 ± 337 vs. 967 ± 217 μg/ml, p=0.05). Samples of 15-μg were subjected to 2D gel electrophoresis. After staining with SYPRO-Ruby protein stein, spot intensities were determined using image analysis software. Clearly visible spots were excised from the gel, digested in situ with trypsin, and the proteins were identified by liquid chromatography tandem MS (LC MS/MS) sequence analysis. We identified 8 spots that were markedly higher in DME than non-DME samples. From the 8 spots, 6 proteins were identified, i.e.PEDF, Apo A-4, ApoA-1, Trip-11, PRBP, and vitamin D-binding protein. One spot was expressed only in non-DME, and it was identified Apo-H. We posit that these chemical mediators presence in the posterior vitreous may play a role in the pathogenesis of DME.