4-Acetoxyphenol Prevents RPE Oxidative Stress-Induced Necrosis by Functioning as an NRF2 Stabilizer
4-Acetoxyphenol Prevents RPE Oxidative Stress-Induced Necrosis by Functioning as an NRF2 Stabilizer
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DOI：
10.1167/iovs.15-16401
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发表时间：
2015-08-01
影响因子：
4.4
通讯作者：
Wang, Shusheng
中科院分区：
文献类型：
--
作者：
Hanus, Jakub;Kolkin, Alexander;Wang, Shusheng
目的。氧化应激已被认为是AMD发病的主要危险因素。视网膜色素上皮细胞（RPE）对维持视网膜的稳态至关重要，在干性AMD中，特别是在地理萎缩中，RPE细胞死亡和由此产生的光感受器凋亡已被观察到。本文的目的是鉴定和重新利用美国食品药品监督管理局批准的天然化合物4-乙酰氧基酚（4-AC），并评价其对氧化应激诱导的RPE坏死的保护作用和机制。我们用4-AC预处理ARPE-19细胞后，将其暴露于过氧化叔丁基（tBHP）中，用MTT法测定细胞活力。通过转染的报告基因分析RIPK3和HMGB1核释放的聚集情况。使用市售的活性氧检测系统测量活性氧（ROS）。通过siRNA研究、qRT-PCR和免疫染色证实了NRF2/NQO1/HO-1通路在介导4-AC功能中的重要性。我们已经确定了一种天然抗氧化剂，4-AC，它具有很强的保护RPE细胞免受氧化应激诱导的坏死的能力。机制上，4-AC通过稳定和诱导NRF2转录因子核易位，阻断氧化应激诱导的细胞ROS增加，上调NQO1和HO-1基因。NQO1、HO-1和NRF2进一步被证明是4-AC保护RPE细胞免受tBHP诱导死亡所必需的。NRF2稳定剂thbhq与4-AC对thbhq诱导的RPE死亡的保护作用一致。化合物4-AC通过稳定NRF2，上调NQO1和HO-1基因，保护ARPE-19细胞免受氧化应激诱导的坏死。
PURPOSE. Oxidative stress has been suggested to be a major risk factor for the pathogenesis of AMD. Retinal pigment epithelial (RPE) cells are essential for maintaining the homeostasis of the retina, and RPE cell death and the resultant photoreceptor apoptosis have been observed in dry AMD, especially in geographic atrophy. The purpose of this article was to identify and repurpose the Food and Drug Administration-approved natural compound 4-Acetoxyphenol (4-AC), and to evaluate its effect and mechanism in protecting against oxidative stress-induced RPE necrosis.METHODS. We exposed ARPE-19 cells to tert-Butyl hydroperoxide (tBHP) after pretreatment with 4-AC, and measured cell viability by MTT assay. Aggregation of RIPK3 and HMGB1 nuclear release were analyzed by transfected reporter genes. Reactive oxygen species (ROS) were measured using a commercially available ROS detection system. The importance of the NRF2/NQO1/HO-1 pathway in mediating 4-AC function was corroborated by siRNA studies, qRT-PCR, and immunostaining.RESULTS. We have identified a natural antioxidant, 4-AC, which demonstrates strong abilities to protect RPE cells from oxidative stress-induced necrosis. Mechanistically, 4-AC blocked the increase of cellular ROS induced by oxidative stress, and upregulated NQO1 and HO-1 genes by stabilizing and inducing the nuclear translocation of NRF2 transcription factor. The NQO1, HO-1, and NRF2 were further shown to be required for 4-AC protection of RPE cells from death induced by tBHP. The tBHQ, an NRF2 stabilizer, consistently mimicked the protective effect of 4-AC against tBHP-induced RPE death.CONCLUSIONS. The compound 4-AC protects ARPE-19 cells from oxidative stress-induced necrosis through upregulation of NQO1 and HO-1 genes by stabilization of NRF2.