半乳糖凝集素-1 在增殖性糖尿病视网膜病变中的研究。

Galectin-1 studies in proliferative diabetic retinopathy.

机构信息

Department of Ophthalmology, King Saud University, Riyadh, Saudi Arabia.

Dr. Nasser Al-Rashid Research Chair in Ophthalmology College of Medicine, King Saud University, Riyadh, Saudi Arabia.

出版信息

Acta Ophthalmol. 2020 Feb;98(1):e1-e12. doi: 10.1111/aos.14191. Epub 2019 Jul 18.

DOI:10.1111/aos.14191

PMID:31318490

Abstract

PURPOSE

Galectin-1 regulates endothelial cell function and promotes angiogenesis. We investigated the hypothesis that galectin-1 may be involved in the pathogenesis of proliferative diabetic retinopathy (PDR).

METHODS

Vitreous samples from 36 PDR and 20 nondiabetic patients, epiretinal fibrovascular membranes from 13 patients with PDR, rat retinas and human retinal Müller glial cells were studied by enzyme-linked immunosorbent assay (ELISA), immunohistochemistry and Western blot analysis. In vitro angiogenesis assays were performed and the adherence of leukocytes to galectin-1-stimulated human retinal microvascular endothelial cells (HRMECs) was assessed.

RESULTS

The ELISA analysis revealed that galectin-1 and vascular endothelial growth factor (VEGF) levels were significantly higher in vitreous samples from PDR patients than in those from nondiabetics (p < 0.001 for both comparisons). A significant positive correlation was found between the levels of galectin-1 and VEGF (r = 0.354; p = 0.022). In epiretinal membranes, immunohistochemical analysis showed that galectin-1 was expressed in vascular endothelial cells expressing CD31, myofibroblasts expressing α-smooth muscle actin and leukocytes expressing CD45. The galectin-1 receptor neuropilin-1 was expressed on vascular endothelial cells. CD31 staining was used as a marker to assess microvessel density (MVD). Significant positive correlation was detected between MVD in epiretinal membranes and the number of blood vessels expressing galectin-1 (r = 0.848; p < 0.001). Western blot analysis demonstrated significant increase of galectin-1 protein in rat retinas after induction of diabetes. ELISA analysis revealed that hydrogen peroxide and cobalt chloride (CoCl ) induced upregulation of galectin-1 in Müller cells. Treatment with galectin-1 induced upregulation of VEGF in Müller cells and increased leukocyte adhesion to HRMECs. The galectin-1 inhibitor OTX008 attenuated VEGF-induced HRMECs migration and CoCl -induced upregulation of NF-κB, galectin-1 and VEGF in Müller cells.

CONCLUSIONS

These results suggest that galectin-1is involved in the pathogenesis of PDR.

摘要

目的

半乳糖凝集素-1 调节内皮细胞功能并促进血管生成。我们研究了半乳糖凝集素-1 可能参与增生性糖尿病视网膜病变（PDR）发病机制的假说。

方法

通过酶联免疫吸附试验（ELISA）、免疫组织化学和 Western blot 分析，研究了 36 例 PDR 患者和 20 例非糖尿病患者的玻璃体样本、13 例 PDR 患者的视网膜内纤维血管膜、大鼠视网膜和人视网膜 Müller 胶质细胞。进行了体外血管生成试验，并评估了白细胞对半乳糖凝集素-1 刺激的人视网膜微血管内皮细胞（HRMEC）的黏附作用。

结果

ELISA 分析显示，PDR 患者玻璃体样本中的半乳糖凝集素-1 和血管内皮生长因子（VEGF）水平明显高于非糖尿病患者（两者比较均 p<0.001）。发现半乳糖凝集素-1 和 VEGF 水平之间存在显著正相关（r=0.354；p=0.022）。在视网膜内纤维血管膜中，免疫组织化学分析显示，半乳糖凝集素-1 表达于表达 CD31 的血管内皮细胞、表达 α-平滑肌肌动蛋白的肌成纤维细胞和表达 CD45 的白细胞中。半乳糖凝集素-1 受体神经纤毛蛋白-1 表达于血管内皮细胞上。用 CD31 染色作为标记物评估视网膜内纤维血管膜中的微血管密度（MVD）。发现视网膜内纤维血管膜中的 MVD 与表达半乳糖凝集素-1 的血管数量之间存在显著正相关（r=0.848；p<0.001）。Western blot 分析显示，糖尿病诱导后大鼠视网膜中半乳糖凝集素-1 蛋白显著增加。ELISA 分析显示，过氧化氢和氯化钴（CoCl ）诱导 Müller 细胞中半乳糖凝集素-1 的上调。用半乳糖凝集素-1 处理诱导 Müller 细胞中 VEGF 的上调，并增加白细胞与 HRMEC 的黏附。半乳糖凝集素-1 抑制剂 OTX008 减弱了 VEGF 诱导的 HRMEC 迁移和 CoCl 诱导的 Müller 细胞中 NF-κB、半乳糖凝集素-1 和 VEGF 的上调。