中文摘要：

背景脉管的 endothelial 生长因素(VEGF ) 在糖尿病涉及网膜的脉管的漏和 nonperfusion 的开始。细胞内部的粘附 molecule-1 (ICAM-1 ) 是网膜的 leukostasis 上的 VEGF 的效果的关键调停人。尽管 VEGF 在早阶段的糖尿病的视网膜被表示，是否它直接在网膜的 endothelial 房间(EC ) 的起来调整 ICAM-1 是未知的。在这研究，我们提供了新机制解释 VEGF 在网膜的 ECs.Methods 牛的网膜的 EC (BREC ) 做起来调整 ICAM-1 的表示被孤立并且有教养。染色的 Immunohistochemical 被执行识别 BREC。有教养的房间被划分成相应的组。然后， VEGF (100 ng/ml ) 和另外的禁止者被用来对待房间。房间 lysate 和有教养的上层清液被收集，然后， ICAM-1 的蛋白质水平和 endothelial 的 phosphorylation 氮的氧化物 synthase (eNOS ) 用西方的弄污被检测。Griess 反应被用来检测氮的氧化物(没有) 西方的弄污显示出的 .Results VEGF 起来调整 ICAM-1 蛋白质的表示和在网膜的 EC 的 eNOS 的增加的 phosphorylation。既不块不，蛋白质 kinase C (PKC ) 也没改变 ICAM-1 的表示或 eNOS 的 phosphorylation。西方的弄污的结果也显示出 phosphatidylinositol 3-kinase (PI3K ) 的那抑制或反应的氧种类(ROS ) 显著地减少了 ICAM-1 的表示。PI3K 的抑制也减少了 eNOS 的 phosphorylation。Griess 反应显示出显著地在没有生产期间增加的那 VEGF。 eNOS 什么时候被L名字或 PI3K 堵住，被 LY294002 堵住，没有生产的基础水平和增长不由 VEGF 引起了能是显著地 decreased.Conclusion ROS --在网膜的内皮细胞层联合都可以不是能帮助解释 VEGF 为什么在糖尿病的 retinopathy 导致 ICAM-1 表示和产生 leukostasis 的新机制。

英文摘要：

Background The vascular endothelial growth factor （VEGF） is involved in the initiation of retinal vascular leakage and nonperfusion in diabetes. The intracellular adhesion molecule-1 （ICAM-1） is the key mediator of the effect of VEGFs on retinal leukostasis. Although the VEGF is expressed in an early-stage diabetic retina, whether it directly up-regulates ICAM-1 in retinal endothelial cells （ECs） is unknown. In this study, we provided a new mechanism to explain that VEGF does up-regulate the expression of ICAM-1 in retinal ECs. Methods Bovine retinal ECs （BRECs） were isolated and cultured. Immunohistochemical staining was performed to identify BRECs. The cultured cells were divided into corresponding groups. Then, VEGF （100 ng/ml） and other inhibitors were used to treat the cells. Cell lysate and the cultured supernatant were collected, and then, the protein level of ICAM-1 and phosphorylation of the endothelial nitric oxide synthase （eNOS） were detected using Western blotting. Griess reaction was used to detect nitric oxide （NO）. Results Western blotting showed that the VEGF up-regulated the expression of ICAM-1 protein and increased phosphorylation of the eNOS in retinal ECs. Neither the block of NO nor protein kinase C （PKC） altered the expression of ICAM-1 or the phosphorylation of eNOS. The result of the Western blotting also showed that inhibition of phosphatidylinositol 3-kinase （PI3K） or reactive oxygen species （ROS） significantly reduced the expression of ICAM-1. Inhibition of PI3K also reduced phosphorylation of eNOS. Griess reaction showed that VEGF significantly increased during NO production. When eNOS was blocked by L-NAME or PI3K was blocked by LY294002, the basal level of NO production and the increment of NO caused by VEGF could be significantly decreased. Conclusion ROS-NO coupling in the retinal endothelium may be a new mechanism that could help to explain why VEGF induces ICAM-1 expression and the resulting leukostasis in diabetic retinopathy.