Osteoprotegerin Is a New Regulator of Inflammation and Angiogenesis in Proliferative Diabetic Retinopathy
Osteoprotegerin Is a New Regulator of Inflammation and Angiogenesis in Proliferative Diabetic Retinopathy
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DOI：
10.1167/iovs.16-20993
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发表时间：
2017-06-01
影响因子：
4.4
通讯作者：
Simo, Rafael
中科院分区：
文献类型：
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作者：
Abu El-Asrar, Ahmed M.;Struyf, Sofie;Simo, Rafael
目的.骨保护素（OPG）是一种新型的内皮细胞功能，血管生成和血管生成的调节剂。我们将增生性糖尿病视网膜病变（PDR）中OPG的表达水平与血管生成和炎症因子血管内皮生长因子（VEGF）和单核细胞趋化蛋白-1（MCP-1/CCL 2）的表达水平相关联。我们还检测了OPG在糖尿病大鼠和糖尿病患者视网膜中的表达，并测量了人视网膜微血管内皮细胞（HRMEC）的OPG产生，并研究了其血管生成活性。采用酶联免疫吸附试验、免疫组化、免疫荧光、Western印迹分析和RT-PCR研究了47例PDR和28例非糖尿病患者的玻璃体样本、14例PDR患者的视网膜前膜、人视网膜（10例糖尿病患者和10例非糖尿病受试者）、大鼠视网膜和HRMEC。进行体外和体内血管生成测定。我们发现，PDR患者玻璃体样本与非糖尿病对照组相比，OPG、VEGF和MCP-1/CCL2的表达显著增加。OPG水平与VEGF和MCP-1/CCL2水平呈显著正相关。在视网膜前膜中，OPG表达于血管内皮细胞和基质细胞。OPG mRNA和蛋白在糖尿病患者视网膜中的表达明显增加。促炎细胞因子TNF-α和IL-β，而不是VEGF，MCP-1/CCL2或凝血酶，诱导HRMEC中OPG的上调。骨保护素诱导HRMEC中ERK 1/2和Akt磷酸化并刺激其迁移。在体内蛋白质明胶塞试验中，骨保护素增强VEGF的血管生成作用。这些结果表明OPG参与了PDR血管生成。
PURPOSE. Osteoprotegerin (OPG) is a novel regulator of endothelial cell function, angiogenesis, and vasculogenesis. We correlated expression levels of OPG with those of the angiogenic and inflammatory factors vascular endothelial growth factor (VEGF) and monocyte chemoattractant protein-1 (MCP-1/CCL2) in proliferative diabetic retinopathy (PDR). We also examined expression of OPG in retinas from diabetic rats and diabetic patients and measured production of OPG by human retinal microvascular endothelial cells (HRMEC) and investigated its angiogenic activity.METHODS. Vitreous samples from 47 PDR and 28 nondiabetic patients, epiretinal membranes from 14 patients with PDR, human retinas (10 from diabetic patients and 10 from nondiabetic subjects), and rat retinas and HRMEC were studied by using enzyme-linked immunosorbent assay, immunohistochemistry, immunofluorescence, Western blot analysis, and RT-PCR. In vitro and in vivo angiogenesis assays were performed.RESULTS. We showed a significant increase in the expression of OPG, VEGF, and MCP-1/CCL2 in a comparison between vitreous samples from PDR patients and those from nondiabetic controls. Significant positive correlations were found between levels of OPG and levels of VEGF and MCP-1/CCL2. In epiretinal membranes, OPG was expressed in vascular endothelial cells and stromal cells. Significant increases of OPG mRNA and protein were detected in the retinas from diabetic patients. The proinflammatory cytokines TNF-alpha and IL-beta, but not VEGF, MCP-1/CCL2 or thrombin, induced upregulation of OPG in HRMEC. Osteoprotegerin induced ERK1/2 and Akt phosphorylation in HRMEC and stimulated their migration. Osteoprotegerin potentiated the angiogenic effect of VEGF in the in vivo protein gelatin plug assay.CONCLUSIONS. These results suggest that OPG is involved in PDR angiogenesis.