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Quercetin protects ARPE-19 cells against photic stress mediated by the products of rhodopsin photobleaching.
Photochemical & Photobiological Sciences ( IF 2.7 ) Pub Date : 2020-06-08 , DOI: 10.1039/d0pp00165a Magdalena M Olchawa 1 , Olga I Krzysztynska-Kuleta 1 , Krystian T Mokrzynski 1 , Piotr M Sarna 2 , Tadeusz J Sarna 1
Photochemical & Photobiological Sciences ( IF 2.7 ) Pub Date : 2020-06-08 , DOI: 10.1039/d0pp00165a Magdalena M Olchawa 1 , Olga I Krzysztynska-Kuleta 1 , Krystian T Mokrzynski 1 , Piotr M Sarna 2 , Tadeusz J Sarna 1
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Although the primary biological function of retina photoreceptors is to absorb light and provide visual information, exposure to intense light could increase the risk of phototoxic reactions mediated by rhodopsin photobleaching products (RPBP) that might accumulate in photoreceptor outer segments (POS). Here we investigated whether quercetin can modify the phototoxic potential of RPBP under in vitro photic stress conditions. ARPE-19 cells or quercetin enriched cultures pre-loaded with rhodopsin-rich POS isolated from bovine retinas were irradiated with green light to photobleach rhodopsin, and subsequently with blue light. Survival of cells was determined by MTT assay and propidium iodide staining. Changes in mitochondrial membrane potential (MMP) were assessed by JC-1 staining. Protein hydroperoxides, formed by photosensitized oxidation, mediated by RPBP, were analyzed in cells and in a model system with bovine serum albumin (BSA), using the coumarin boronic acid fluorogenic probe. The effect of photic stress on specific phagocytosis of RPE cells was determined by flow cytometry. Photoreactivity of POS with and without quercetin was analyzed by EPR oximetry and EPR spin trapping. Cytotoxicity measurements and MMP analyses confirmed that supplementation with quercetin protected ARPE-19 cells against photic stress mediated by rhodopsin-rich POS. Quercetin significantly reduced the inhibitory effect of RPBP-mediated stress on POS phagocytosis and the RPBP ability to photooxidize cellular proteins or BSA. The data support the hypothesis that quercetin may efficiently diminish the phototoxic action of retinoids, necessary for restoring the phagocytic function of ARPE-19 cells.
中文翻译：
槲皮素可保护ARPE-19细胞免受视紫红质光漂白产物介导的光胁迫。
尽管视网膜感光器的主要生物学功能是吸收光并提供视觉信息，但是暴露于强光下可能会增加视紫红质光漂白产品（RPBP）介导的光毒反应的风险，而视紫红质光漂白产品（RPBP）可能会在感光器外段（POS）中积累。在这里我们调查槲皮素是否可以在体外改变RPBP的光毒性潜力逆境条件。预加载从牛视网膜分离的富含视紫红质的POS的ARPE-19细胞或富含槲皮素的培养物，先用绿光照射视紫红质，然后用蓝光对它们进行漂白。通过MTT测定和碘化丙啶染色确定细胞的存活。通过JC-1染色评估线粒体膜电位（MMP）的变化。由RPBP介导的光敏氧化形成的蛋白质氢过氧化物，使用香豆素硼酸荧光探针在牛血清白蛋白（BSA）的细胞和模型系统中进行了分析。通过流式细胞术确定光胁迫对RPE细胞特异性吞噬作用的影响。有和没有槲皮素的POS的光反应性通过EPR血氧测定法和EPR自旋阱分析。细胞毒性测量和MMP分析证实，补充槲皮素可保护ARPE-19细胞免受富含视紫红质的POS介导的光胁迫。槲皮素显着降低RPBP介导的应激对POS吞噬作用的抑制作用以及RPBP光氧化细胞蛋白或BSA的能力。数据支持以下假设：槲皮素可有效减少类视色素的光毒性作用，这是恢复ARPE-19细胞吞噬功能所必需的。
更新日期：2020-08-12
中文翻译：
槲皮素可保护ARPE-19细胞免受视紫红质光漂白产物介导的光胁迫。
尽管视网膜感光器的主要生物学功能是吸收光并提供视觉信息，但是暴露于强光下可能会增加视紫红质光漂白产品（RPBP）介导的光毒反应的风险，而视紫红质光漂白产品（RPBP）可能会在感光器外段（POS）中积累。在这里我们调查槲皮素是否可以在体外改变RPBP的光毒性潜力逆境条件。预加载从牛视网膜分离的富含视紫红质的POS的ARPE-19细胞或富含槲皮素的培养物，先用绿光照射视紫红质，然后用蓝光对它们进行漂白。通过MTT测定和碘化丙啶染色确定细胞的存活。通过JC-1染色评估线粒体膜电位（MMP）的变化。由RPBP介导的光敏氧化形成的蛋白质氢过氧化物，使用香豆素硼酸荧光探针在牛血清白蛋白（BSA）的细胞和模型系统中进行了分析。通过流式细胞术确定光胁迫对RPE细胞特异性吞噬作用的影响。有和没有槲皮素的POS的光反应性通过EPR血氧测定法和EPR自旋阱分析。细胞毒性测量和MMP分析证实，补充槲皮素可保护ARPE-19细胞免受富含视紫红质的POS介导的光胁迫。槲皮素显着降低RPBP介导的应激对POS吞噬作用的抑制作用以及RPBP光氧化细胞蛋白或BSA的能力。数据支持以下假设：槲皮素可有效减少类视色素的光毒性作用，这是恢复ARPE-19细胞吞噬功能所必需的。
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