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The application of diagnosis and treatment of Angiopoietin and Ephrin from retinal ischemia in the angiogenesis

血管生成素和Ephrin在视网膜缺血血管新生诊治中的应用

基本信息

批准号：
12671696
负责人：
KISHI Shouji
金额：
$ 2.18万
依托单位：
Gunma University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

项目摘要

Angiogenesis produces retinal ischemia by circulatory disturbance of retinal vessel. Many cytokines such as vascular endothelial grouth factor (VGEF) seem to be related to the angiogenesis. In order to induce rubeosis and neovascularization secondary to retinal ischemia, major retinal veins were occluded with laser photocoagulation in 6 eyes of 3 Japanese monkeys. Two eyes with rubeosis and neovascularization were enucleated one week, two weeks and four weeks respectively after photocoagulation. Immunohistochemical analysis was performed on fixed paraffin sections of the iris, trabeculum and retinal neovascularization using polyclonal antibody against Ephrin-B2, Eph-B4, Angiopoietin-2 and VEGF. Antibody localization was examined using anavidin-biotin reaction linked to aminoethylcarbabazole color reagent the trabecular meshwork and the Schlemms canal including 1, 2 and 4 weeks after treatment, showed positive staining for VEGF. Ephrin-B2 was positively stained in the section of 2 and 4 weeks after treatment but it was uncertain in the sections of one week. Ephrin- B4 and Angiopoietin-2 were not significant in any sections. All retinal neovascularization 1, 2 and 4 weeks after treatment, showed positive staining for VEGF. One case, Ephrin-B2, B4 and Angiopoietin-2 were positively stained in the section for only 2 weeks after treatment, but other case was uncertain for Ephrin- B4 and Angiopoietin-2. It was uncertain in the sections for one and four weeks. Expression of VEGF was followed by that of Ephrin-B2 in experimentally induced rubeosis and neovasucularization. The data suggests the expression of Ephrin-B2, an arterial differentiation, might play a role in grouth of newly formed vessels.

血管新生通过视网膜血管循环障碍引起视网膜缺血。许多细胞因子如血管内皮生长因子（VGEF）与血管生成有关。为了诱导继发于视网膜缺血的发红和新生血管形成，在3只日本猴的6只眼睛中用激光光凝法阻塞主要视网膜静脉。分别于光凝后1周、2周、4周行眼球摘除术。免疫组织化学分析进行固定石蜡切片的虹膜，小梁和视网膜新生血管，使用多克隆抗体Ephrin-B2，Eph-B4，血管生成素-2和VEGF。使用与氨乙基卡巴唑显色试剂连接的抗生物素蛋白-生物素反应检查抗体定位，治疗后1周、2周和4周的小梁网和施莱姆氏管均显示VEGF阳性染色。Ephrin-B2在治疗后2周和4周的切片中呈阳性染色，但在治疗后1周的切片中染色不确定。Ephrin-B4和Angiopoietin-2在任何切片中均不显著。治疗后1、2、4周，所有视网膜新生血管均显示VEGF阳性染色。1例治疗后2周Ephrin-B2、B4和Angiopoietin-2染色阳性，另1例Ephrin-B4和Angiopoietin-2染色不确定。这是不确定的部分一个星期和四个星期。在实验诱导的红斑和新生血管形成中，Ephrin-B2的表达紧随VEGF的表达。这些数据表明Ephrin-B2的表达，动脉分化，可能在新形成的血管的生长中发挥作用。