Decorin Prevents Retinal Pigment Epithelial Barrier Breakdown Under Diabetic Conditions by Suppressing p38 MAPK Activation
Decorin Prevents Retinal Pigment Epithelial Barrier Breakdown Under Diabetic Conditions by Suppressing p38 MAPK Activation
复制标题
核心蛋白聚糖通过抑制 p38 MAPK 激活来防止糖尿病患者视网膜色素上皮屏障的破坏
DOI：
10.1167/iovs.14-15874
复制
发表时间：
2015-05-01
影响因子：
4.4
通讯作者：
Li, Tingting
中科院分区：
文献类型：
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作者：
Wang, Shuai;Du, Shanshan;Li, Tingting
目的。本研究旨在探讨在高糖(HG) +缺氧条件下，decorin对人视网膜色素上皮（RPE）细胞屏障功能的影响。人RPE （ARPE-19）细胞在正常葡萄糖（5.5 mM）或HG （25 mM）培养基中培养18天。此外，为了模拟糖尿病视网膜病变中发生的缺氧影响，在实验的最后2天，用100 μ M的CoCl2处理细胞。在加入CoCl2 1小时前，应用Decorin， 100 nM。通过测定异硫氰酸荧光素-葡聚糖的经皮电阻（TER）和顶-基底侧渗透性来评价视网膜色素上皮屏障功能。Western blotting和免疫荧光法检测紧密连接蛋白（claudin-1、occludin和zonula occludens-1 [ZO-1]）的含量和分布。Western blotting检测p38丝裂原活化蛋白激酶（MAPK）磷酸化水平，并在ARPE-19单层中转染小干扰RNA。高糖加缺氧显著增加fitc -葡聚糖通透性，并伴有TER降低。Decorin逆转了这两种效应。高糖加缺氧诱导的occludin和ZO-1的减少和紊乱也被decorin逆转。Decorin可抑制缺氧诱导的p38 MAPK活化。RNA干扰p38 MAPK沉默也能抑制HG +缺氧诱导的ARPE-19细胞单层的破坏。decorin通过抑制p38 MAPK的激活来阻止HG +缺氧诱导的视网膜色素上皮屏障破坏，这可能为抑制糖尿病黄斑水肿提供一种新的治疗策略。
PURPOSE. The purpose of this study was to determine the effect of decorin on the barrier function of human retinal pigment epithelial (RPE) cells under high-glucose (HG) plus hypoxia conditions.METHODS. Human RPE (ARPE-19) cells were cultured for 18 days in normal glucose (5.5 mM) or HG (25 mM) medium. In addition, to mimic the hypoxic impact which occurs in diabetic retinopathy, cells were treated with 100 mu M CoCl2 during the last 2 days of the experiment. Decorin, 100 nM, was applied 1 hour before CoCl2 was added. Retinal pigment epithelial barrier function was evaluated by measuring transepithelial electrical resistance (TER) and apical-basolateral permeability of fluorescein isothiocyanate (FITC)-dextran. The content and distribution of tight junction proteins (claudin-1, occludin, and zonula occludens-1 [ZO-1]) were examined by Western blotting and immunofluorescence. p38 mitogen-activated protein kinase (MAPK) phosphorylation was evaluated by Western blotting, and small interfering RNA transfection to p38 MAPK was also performed in ARPE-19 monolayers.RESULTS. High-glucose plus hypoxia significantly increased FITC-dextran permeability, paralleled by decreased TER. Decorin reversed both of these effects. High-glucose plus hypoxia-induced reduction and disorganization of occludin and ZO-1 were also reversed by decorin. Decorin prevented the activation of p38 MAPK induced by hypoxia. Silence of p38 MAPK by RNA interference also inhibited the breakdown of ARPE-19 cell monolayer induced by HG plus hypoxia.CONCLUSIONS. Retinal pigment epithelial barrier disruption induced by HG plus hypoxia was prevented by decorin through suppression of p38 MAPK activation, which could present a new therapeutic strategy for inhibition of diabetic macular edema.