Ouchi Masayuki, West Karen, Crabb John W, Kinoshita Shigeru, Kamei Motohiro
Department of Ophthalmology, Kyoto Prefectural University of Medicine, 465 Kawaramachi Hirokoji Kamigyo-ku, Kyoto 602-0841, Kyoto, Japan.
Exp Eye Res. 2005 Aug;81(2):176-82. doi: 10.1016/j.exer.2005.01.020.
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 were corrected from 20 eyes with pre-proliferative diabetic retinopathy associated with DME (DME group) and without DME (non-DME group). They were subjected to 2D gel electrophoresis, and the spot intensities were compared between the groups. Apparently visible spots were excised from the gel, and the proteins were identified by liquid chromatography tandem MS (LC MS/MS) sequence analysis. We identified 14 proteins from the DME group, and 15 proteins from the non-DME group. The intensity of eight spots was markedly higher in DME than non-DME samples and one spot was detected only in non-DME samples. From the eight spots, six proteins were identified, including PEDF, ApoA-4, ApoA-1, Trip-11, PRBP, and VDBP. On the other hand, Apo H was expressed only in non-DME. Certain vitreous proteins expressed exclusively in DME and lacked in DME. These chemical mediators in the posterior vitreous may play a role in the pathogenesis of DME.
使用二维凝胶（2D凝胶）电泳和质谱（MS）技术，整体鉴定和分析玻璃体内与糖尿病性黄斑水肿（DME）相关的蛋白质。从20只患有与DME相关的增殖前期糖尿病视网膜病变的眼睛（DME组）和未患DME的眼睛（非DME组）采集玻璃体液样本。对样本进行2D凝胶电泳，并比较两组之间的斑点强度。从凝胶中切下明显可见的斑点，通过液相色谱串联质谱（LC MS/MS）序列分析鉴定蛋白质。我们从DME组中鉴定出14种蛋白质，从非DME组中鉴定出15种蛋白质。DME样本中8个斑点的强度明显高于非DME样本，并且仅在非DME样本中检测到1个斑点。从这8个斑点中鉴定出6种蛋白质，包括色素上皮衍生因子（PEDF）、载脂蛋白A-4（ApoA-4）、载脂蛋白A-1（ApoA-1）、Trip-11、孕激素受体结合蛋白（PRBP）和维生素D结合蛋白（VDBP）。另一方面，载脂蛋白H（Apo H）仅在非DME中表达。某些玻璃体内蛋白质仅在DME中表达而在非DME中缺乏。玻璃体后部的这些化学介质可能在DME的发病机制中起作用。
