Fenofibric acid reduces fibronectin and collagen type IV overexpression in human retinal pigment epithelial cells grown in conditions mimicking the diabetic milieu: functional implications in retinal permeability.
Fenofibric acid reduces fibronectin and collagen type IV overexpression in human retinal pigment epithelial cells grown in conditions mimicking the diabetic milieu: functional implications in retinal permeability.
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DOI：
10.1167/iovs.11-7282
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发表时间：
2011-08
影响因子：
4.4
通讯作者：
K. Trudeau;Sumon Roy;Wen Guo;C. Hernández;M. Villarroel;R. Simó;Sayon Roy
中科院分区：
文献类型：
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作者：
K. Trudeau;Sumon Roy;Wen Guo;C. Hernández;M. Villarroel;R. Simó;Sayon Roy
目的探讨非诺纤维酸（FA）是否能降低高糖（HG）诱导的人视网膜色素上皮（RPE）细胞基底膜组分的过表达和高通透性。方法将视网膜色素上皮细胞（ARPE-19）在正常葡萄糖（5 mM）或HG （25 mM）培养基中培养18 d，观察FA对纤维连接蛋白（FN）和胶原蛋白IV （Coll IV）表达的影响。在实验的最后3天，在HG培养基或HG培养基加IL-1β (HG + IL-1β)中培养的细胞中添加100 μM FA，至少部分模拟糖尿病环境的炎症方面。Real-time RT-PCR检测FN和Coll IV mRNA水平，Western blot检测蛋白水平。使用针对紧密连接蛋白ZO-1和claudin-1的特异性抗体，以及通过测量fitc -葡聚糖的顶端-基底外侧运动，通过共聚焦显微镜分析细胞单层形态和屏障功能。结果HG和HG + IL-1β培养的RPE细胞FN和Coll IV表达明显高于正常培养基。在HG或HG + IL-1β培养基中生长的细胞经FA处理后，FN和Coll IV的表达显著降低。此外，在HG + IL-1β培养基中生长的细胞中，暴露于FA以剂量依赖的方式降低了过度通透性。这种影响与紧密连接蛋白含量的变化无关。结论：本研究结果表明，FA下调基底膜成分可能对糖尿病视网膜病变相关的外血-视网膜屏障渗漏具有保护作用。
PURPOSE To determine whether fenofibric acid (FA) reduces high glucose (HG)-induced basement membrane component overexpression and hyperpermeability in human retinal pigment epithelial (RPE) cells. METHODS Retinal pigment epithelial cells (ARPE-19) were cultured for 18 days in normal glucose (5 mM) or HG (25 mM) medium and studied for the effects of FA on fibronectin (FN) and collagen IV (Coll IV) expression. During last 3 days of the experiment, 100 μM FA was added to cells grown in HG medium or in HG medium plus IL-1β (HG + IL-1β) to mimic, at least in part, the inflammatory aspect of the diabetic milieu. Real-time RT-PCR was performed to determine FN and Coll IV mRNA levels, whereas protein levels were assessed by Western blot analyses. Cell monolayer morphology and barrier function were analyzed by confocal microscopy using specific antibodies against tight junction proteins, ZO-1, and claudin-1 and by measuring apical-basolateral movements of FITC-dextran, respectively. RESULTS FN and Coll IV expression were significantly increased in RPE cells grown in HG or HG + IL-1β medium compared with cells grown in normal medium. When cells grown in HG or HG + IL-1β medium were treated with FA, significant reductions in FN and Coll IV expression were observed. In addition, exposure to FA decreased excess permeability in a dose-dependent manner in cells grown in HG + IL-1β medium. This effect was unrelated to changes in tight junction protein content. CONCLUSIONS Findings from this study suggest that the downregulation of basement membrane components by FA may have a protective effect against outer blood-retinal barrier leakage associated with diabetic retinopathy.