RAGE Is Essential for Subretinal Fibrosis in Laser-Induced Choroidal Neovascularization: Therapeutic Implications.

IF 4.7 2区 医学 Q1 OPHTHALMOLOGY
Parameswaran G Sreekumar, Mi-Hyun Nam, Elise Hong, Ram Kannan, Ram H Nagaraj
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引用次数: 0

Abstract

Purpose: Subretinal fibrosis, a complication of neovascular age-related macular degeneration (nAMD), involves the epithelial-mesenchymal transition (EMT) of retinal pigment epithelium (RPE) cells as a contributing mechanism. The receptor for advanced glycation end products (RAGE) is a multiligand receptor implicated in fibrotic diseases, but its role in subretinal fibrosis has not been studied. This study investigated the role of RAGE in subretinal fibrosis.

Methods: Subretinal fibrosis was induced in male RAGE-/- and wild-type (WT) mice via laser photocoagulation, and fibrosis lesion volume was assessed on day 35 using optical coherence tomography and immunostaining. In vitro, EMT was induced in primary human RPE cells with transforming growth factor-beta 2 (TGF-β2). The role of RAGE in EMT was studied in cells pretreated with RAGE antagonists (FPS-ZM1 or azeliragon), followed by cotreatment with TGF-β2 for 48 hours. Signaling studies were conducted by pretreatment with FPS-ZM1 for 2 hours, cotreatment with TGF-β2 for 60 minutes, and subsequent immunoblot analysis.

Results: In RAGE-/- mice, subretinal fibrosis after laser-induced choroidal neovascularization was significantly reduced, with a smaller fibrosis volume, less inflammation, decreased activation of pSmad2, and reduced deposition of fibrotic markers (αSMA, collagen I) compared to WT mice. In vitro treatment with TGF-β2 in human RPE cells increased mitochondrial reactive oxygen species and upregulated EMT markers (αSMA, collagen I, and fibronectin), which were inhibited by cotreatment with FPS-ZM1 or azeliragon. FPS-ZM1 blocked TGF-β2-induced Smad2-dependent signaling and EMT without affecting the extracellular signal-regulated kinase (ERK) pathway.

Conclusions: Our findings indicate that RAGE plays a role in RPE cell EMT in subretinal fibrosis and that RAGE antagonists attenuate this process, making RAGE a promising therapeutic target for subretinal fibrosis in nAMD.

RAGE对激光诱导脉络膜新生血管视网膜下纤维化至关重要:治疗意义。
目的:视网膜下纤维化是新生血管性年龄相关性黄斑变性(nAMD)的一种并发症,与视网膜色素上皮(RPE)细胞的上皮-间质转化(EMT)有关。晚期糖基化终产物受体(RAGE)是一种多配体受体,与纤维化疾病有关,但其在视网膜下纤维化中的作用尚未被研究。本研究探讨RAGE在视网膜下纤维化中的作用。方法:采用激光光凝法诱导RAGE-/-和野生型(WT)雄性小鼠视网膜下纤维化,第35天采用光学相干断层扫描和免疫染色法评估纤维化病变体积。在体外,用转化生长因子-β2 (TGF-β2)诱导人RPE原代细胞产生EMT。用RAGE拮抗剂(FPS-ZM1或azeliragon)预处理细胞,然后与TGF-β2共处理48小时,研究RAGE在EMT中的作用。通过FPS-ZM1预处理2小时,与TGF-β2共处理60分钟,进行信号转导研究,随后进行免疫印迹分析。结果:在RAGE-/-小鼠中,与WT小鼠相比,激光诱导脉络膜新生血管形成后视网膜下纤维化明显减少,纤维化体积更小,炎症更少,pSmad2活化降低,纤维化标志物(αSMA,胶原I)沉积减少。TGF-β2在体外处理人RPE细胞时增加了线粒体活性氧,上调了EMT标志物(αSMA,胶原I和纤维连接蛋白),而与FPS-ZM1或阿泽拉贡共处理可抑制这些标志物。FPS-ZM1阻断TGF-β2诱导的smad2依赖性信号通路和EMT,而不影响细胞外信号调节激酶(ERK)通路。结论:我们的研究结果表明,RAGE在视网膜下纤维化的RPE细胞EMT中发挥作用,RAGE拮抗剂减弱这一过程,使RAGE成为nAMD视网膜下纤维化的一个有希望的治疗靶点。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
6.90
自引率
4.50%
发文量
339
审稿时长
1 months
期刊介绍: Investigative Ophthalmology & Visual Science (IOVS), published as ready online, is a peer-reviewed academic journal of the Association for Research in Vision and Ophthalmology (ARVO). IOVS features original research, mostly pertaining to clinical and laboratory ophthalmology and vision research in general.
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