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Quercetin protects retina external barrier from oxidative stress injury by promoting autophagy
Cutaneous and Ocular Toxicology ( IF 1.6 ) Pub Date : 2020-12-17 , DOI: 10.1080/15569527.2020.1860082
Dong Li Li 1, 2, 3, 4, 5 , Lei Mao 6 , Qing Gu 1, 2, 3, 4, 5 , Fang Wei 1, 2, 3, 4, 5 , Yuan-Yuan Gong 1, 2, 3, 4, 5
Affiliation  

Abstract

Purpose: This study aimed to investigate the protective effects of quercetin on the tight junction proteins of human retinal pigment epithelial cells (ARPE-19 cells) suffering from oxidative stress injury and explore the possible mechanism.

Methods: H2O2 (300 μM) was used to establish an oxidative stress model of ARPE-19 cells. ARPE-19 cells were pretreated with different concentrations (0–80 μM) of quercetin before H2O2 exposure. The expression and distribution of tight junction proteins and autophagy-related proteins were detected by Western blot and immunostaining. ARPE-19 cells were pretreated with 5 mM 3-methyladenine (3-MA).

Results: The cell viability weakened in the H2O2 group compared with the control group. However, it was preserved after pretreatment with quercetin. It was observed that the expression levels of occludin, claudin-1 were decreased in the H2O2 group. Quercetin treatment significantly enhanced the expression levels of them as compared to the H2O2 group. H2O2 alone strongly decreased the Zonula occludens protein 1 (ZO-1) expression in the cytomembrane. Quercetin supplementation enhanced the accumulation of ZO-1 in ARPE-19 cells. The expression levels of Beclin-1 and Microtubule associated protein light chain 3 II (LC-3II) increased, and that of P62 decreased in the quercetin protection group. The appearance of LC-3II, which examined by immunofluorescence experiments, enhanced in the quercetin protection group as compared with the control group. The expression levels of beclin-1 and LC-3II increased, and that of P62 increased in the autophagy-inhibited group compared with the quercetin protection group. The levels of occludin and claudin-1 also decreased.

Conclusion: Quercetin prevents the loss of tight junction proteins by upregulating autophagy after oxidative stress in ARPE-19 cells.



中文翻译:

槲皮素通过促进自噬保护视网膜外部屏障免受氧化应激损伤

摘要

目的:本研究旨在探讨槲皮素对人视网膜色素上皮细胞(ARPE-19细胞)氧化应激损伤的紧密连接蛋白的保护作用,并探讨其可能的机制。

方法:采用H 2 O 2 (300 μM)建立ARPE-19细胞氧化应激模型。ARPE-19 细胞在 H 2 O 2暴露之前用不同浓度(0-80 μM)的槲皮素预处理。Western blot和免疫染色检测紧密连接蛋白和自噬相关蛋白的表达和分布。ARPE-19 细胞用 5 mM 3-甲基腺嘌呤 (3-MA) 预处理。

结果:与对照组相比,H 2 O 2组细胞活力减弱。然而,它在用槲皮素预处理后得以保存。观察到H 2 O 2组中occludin、claudin-1的表达水平降低。与H 2 O 2组相比,槲皮素处理显着增强了它们的表达水平。H 2 O 2单独强烈降低细胞膜中封闭带蛋白 1 (ZO-1) 的表达。槲皮素补充剂增强了 ZO-1 在 ARPE-19 细胞中的积累。槲皮素保护组Beclin-1和微管相关蛋白轻链3II(LC-3II)表达水平升高,P62表达水平降低。通过免疫荧光实验检测的LC-3II的出现在槲皮素保护组中与对照组相比增强。与槲皮素保护组相比,自噬抑制组beclin-1和LC-3II表达水平升高,P62表达水平升高。occludin 和 claudin-1 的水平也降低了。

结论:槲皮素通过上调 ARPE-19 细胞氧化应激后的自噬来防止紧密连接蛋白的丢失。

更新日期:2021-02-24
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