Phloroglucinol protects retinal pigment epithelium and photoreceptor against all-trans-retinal-induced toxicity and inhibits A2E formation (original) (raw)
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New Lipophenol Antioxidants Reduce Oxidative Damage in Retina Pigment Epithelial Cells
Antioxidants
Age-related macular degeneration (AMD) is a multifactorial pathology and its progression is exacerbated by oxidative stress. Oxidation and photo-oxidation reactions modify lipids in retinal cells, contribute to tissue injury, and lead to the formation of toxic adducts. In particular, autofluorescent pigments such as N-retinylidene-N-retinylethanolamine (A2E) accumulate as lipofuscin in retinal pigment epithelial cells, contribute to the production of additional reactive oxygen species (ROS), and lead to cell degeneration. In an effort to develop efficient antioxidants to reduce damage caused by lipid oxidation, various natural polyphenols were structurally modified to increase their lipophilicity (lipophenols). In this study, resveratrol, phloroglucinol, quercetin and catechin were selected and conjugated to various polyunsaturated fatty acids (PUFAs) using classical chemical strategies or enzymatic reactions. After screening for cytotoxicity, the capacity of the synthesized lipophe...
New lipophenols prevent carbonyl and oxidative stresses involved in macular degeneration
Free Radical Biology and Medicine, 2021
Dry age-related macular degeneration and Stargardt disease undergo a known toxic mechanism caused by carbonyl and oxidative stresses (COS). This is responsible for accumulation in the retinal pigment epithelium (RPE) of A2E, a main toxic pyridinium bis-retinoid lipofuscin component. Previous studies have shown that carbonyl stress in retinal cells could be reduced by an alkyl-phloroglucinol-DHA conjugate (lipophenol). Here, we performed a rational design of different families of lipophenols to conserve anti-carbonyl stress activities and improve antioxidant properties. Five synthetic pathways leading to alkyl-(poly)phenol derivatives, with phloroglucinol, resveratrol, catechin and quercetin as the main backbone, linked to poly-unsaturated fatty acid, are presented. These lipophenols were evaluated in ARPE-19 cell line for their anti-COS properties and a structureactivity relationship study is proposed. Protection of ARPE-19 cells against A2E toxicity was assessed for the four best candidates. Finally, interesting anti-COS properties of the most promising quercetin lipophenol were confirmed in primary RPE cells.
Isopropyl‐phloroglucinol‐DHA protects outer retinal cells against lethal dose of all‐trans‐retinal
Journal of Cellular and Molecular Medicine, 2020
All‐trans‐retinal (atRAL) is a highly reactive carbonyl specie, known for its reactivity on cellular phosphatidylethanolamine in photoreceptor. It is generated by photoisomerization of 11‐cis‐retinal chromophore linked to opsin by the Schiff's base reaction. In ABCA4‐associated autosomal recessive Stargardt macular dystrophy, atRAL results in carbonyl and oxidative stress, which leads to bisretinoid A2E, accumulation in the retinal pigment epithelium (RPE). This A2E‐accumulation presents as lipofuscin fluorescent pigment, and its photooxidation causes subsequent damage. Here we describe protection against a lethal dose of atRAL in both photoreceptors and RPE in primary cultures by a lipidic polyphenol derivative, an isopropyl‐phloroglucinol linked to DHA, referred to as IP‐DHA. Next, we addressed the cellular and molecular defence mechanisms in commonly used human ARPE‐19 cells. We determined that both polyunsaturated fatty acid and isopropyl substituents bond to phloroglucinol ...
(R)- -Lipoic Acid Protects Retinal Pigment Epithelial Cells from Oxidative Damage
Investigative Ophthalmology & Visual Science, 2005
PURPOSE-To determine whether (R)-α-lipoic acid (LA) protects cultured human fetal retinal pigment epithelial (hfRPE) cells against oxidative injury and identify the pathways that may mediate protection. METHODS-Cultured hfRPE cells were pretreated with various concentrations of LA for 14 to 16 hours followed by treatment with a chemical oxidant, tert-butylhydroperoxide (t-BuOOH; 0.8 mM, 3 hours). Reactive oxygen species (ROS) production and cell viability were measured using H 2 DCF and MTT assays, respectively. RPE cells were evaluated with fluorescent dyes (SYTOX Orange and SYTO Green; Molecular Probes, Eugene, OR), which differentiate between live and dead cells. Apoptosis was visualized by using the TUNEL assay. Changes in mitochondrial membrane potential were detected by JC-1 dye. Intracellular levels of reduced glutathione (GSH) and oxidized glutathione (GSSG) were measured by HPLC. Regulation of γ-glutamylcysteine ligase (GCL), the ratecontrolling enzyme of GSH production, was assayed by RT-PCR. RESULTS-Pretreatment of hfRPE cells with LA, 0.2 mM and 0.5 mM, significantly reduced the levels of t-BuOOH-induced intra-cellular ROS, by 23% and 49%, respectively. LA (0.5 mM) prevented oxidant-induced cell death and apoptosis and also increased the viability of oxidant-treated hfRPE cells from 38% to 90% of control. LA upregulated the mRNA expression of GCL, and was protective against t-BuOOH-induced decreases in both mitochondrial membrane potential and intracellular levels of GSH and GSH/GSSG. CONCLUSIONS-The present study suggests that the protective effect of LA involves multiple pathways and that LA could be effective against age-associated increase in oxidative stress and mitochondrial dysfunction in RPE cells. Age-related macular degeneration (AMD) is the major cause of severe visual impairment for elderly individuals in developed countries. 1-3 Although the underlying pathogenic mechanisms and risk factors are not well understood, 1-9 epidemiologic studies suggest that environmental factors associated with oxidative stress-for example, cigarette smoking, sunlight exposure, and low dietary intake of antioxidants-are contributing factors in the development of AMD. 4-7 Recent results have shown that antioxidants and zinc protect against the progression of the disease. 8
Free Radicals and Antioxidants, 2017
Background: Age-related macular degeneration (AMD) is a leading cause of blindness in the United States among adults age 60 and older. While oxidative stress is implicated in the pathogenesis of AMD, dietary antioxidants have been shown to delay AMD progression in clinical studies. We hypothesized that N-acetylcysteine amide (NACA), a thiol antioxidant, would protect retinal pigment epithelium and impede progression of retinal degeneration. Methods: tert-Butyl hydroperoxide (TBHP) was used to induce oxidative stress in cell cultures. The goal was to evaluate the efficacy of NACA in an in vitro model of AMD in primary human retinal pigment epithelial cells (HRPEpiC). Results: Our data indicates that TBHP generated reactive oxygen species (ROS), which reduced cell viability, depleted glutathione (GSH) levels, and compromised glutathione reductase (GR) activity. Pretreatment with NACA significantly reduced ROS generation, restored GSH levels and GR activity, and recovered transepithelial electrical resistance. Pretreatment with NACA did not decrease the number of dying cells as determined by flow cytometry analysis. However, survival was significantly improved when cells were co-exposed to NACA and TBHP after a shortened pretreatment period. Conclusion: Our data suggest that pretreatment with NACA reduces sublethal but not lethal effects of TBHP in HRPEpiC. NACA significantly improves cell survival when administered prior to and during oxidative damage similar to that observed in the development of dry AMD. These results indicate that continuation of a thiol antioxidant regimen for treatment of AMD is beneficial throughout the course of the disease, and NACA is a potent antioxidant that should be further evaluated for this purpose.
Suppression of Light-Induced Oxidative Stress in the Retina by Mitochondria-Targeted Antioxidant
Antioxidants (Basel, Switzerland), 2018
Light-induced oxidation of lipids and proteins provokes retinal injuries and results in progression of degenerative retinal diseases, such as, for instance, iatrogenic photic maculopathies. Having accumulated over years retinal injuries contribute to development of age-related macular degeneration (AMD). Antioxidant treatment is regarded as a promising approach to protecting the retina from light damage and AMD. Here, we examine oxidative processes induced in rabbit retina by excessive light illumination with or without premedication using mitochondria-targeted antioxidant SkQ1 (10-(6'-plastoquinonyl)decyltriphenyl-phosphonium). The retinal extracts obtained from animals euthanized within 1⁻7 days post exposure were analyzed for H₂O₂, malondialdehyde (MDA), total antioxidant activity (AOA), and activities of glutathione peroxidase (GPx) and superoxide dismutase (SOD) using colorimetric and luminescence assays. Oxidation of visual arrestin was monitored by immunoblotting. The lig...
Experimental Eye Research, 2019
Oxidative cleavage of docosahexaenoate (DHA) in retinal pigmented epithelial (RPE) cells produces 4-hydroxy-7-oxohept-5-enoic acid (HOHA) esters of 2-lysophosphatidylcholine (PC). HOHA-PC spontaneously releases a membrane-permeant HOHA lactone that modifies primary amino groups of proteins and ethanolamine phospholipids to produce 2-(ω-carboxyethyl)pyrrole (CEP) derivatives. CEPs have significant pathological relevance to age-related macular degeneration (AMD) including activation of CEP-specific T-cells leading to inflammatory Ml polarization of macrophages in the retina involved in "dry AMD" and TLR2-dependent induction of angiogenesis that characterizes "wet AMD". RPE cells accumulate DHA from shed rod photoreceptor outer segments through phagocytosis and from plasma lipoproteins secreted by the liver through active uptake from the choriocapillaris. As a cell model of light-induced oxidative damage of DHA phospholipids in RPE cells, ARPE-19 cells were supplemented with DHA, with or without the lipofuscin fluorophore A2E. In this model, light exposure, in the absence of A2E, promoted the generation HOHA lactone-glutathione (GSH) adducts, depletion of intracellular GSH and a competing generation of CEPs. While DHA-rich RPE cells exhibit an inherent proclivity toward light-induced oxidative damage, photosensitization by A2E nearly doubled the amount of lipid oxidation and expanded the spectral range of photosensitivity to longer wavelengths. Exposure of ARPE-19 cells to 1 μM HOHA lactone for 24 h induced massive (50%) loss of lysosomal membrane integrity and caused loss of mitochondrial membrane potential. Using senescence-associated β-galactosidase (SA β-gal) staining that detects lysosomal β-galactosidase, we determined that exposure to HOHA lactone induces senescence in ARPE-19 cells. The present study shows that products of light-induced oxidative damage of DHA phospholipids in the absence of A2E can lead to RPE cell dysfunction. Therefore, their toxicity may be especially important in the early stages of AMD before RPE cells accumulate lipofuscin fluorophores.
Photochemistry and Photobiology, 2007
Throughout the lifetime of an individual, light is focused onto the retina. The resulting photooxidative stress can cause acute or chronic retinal damage. The pathogenesis of age-related macular degeneration (AMD), the leading cause of legal blindness in the developed world, involves oxidative stress and death of the retinal pigment epithelium (RPE) followed by death of the overlying photoreceptors. Evidence suggests that damage due to exposure to light plays a role in AMD and other age-related eye diseases. In this work a system for lightinduced damage and death of the RPE, based on the human ARPE-19 cell line, was used. Induction of mitochondriaderived reactive oxygen species (ROS) is shown to play a critical role in the death of cells exposed to short-wavelength blue light (425 ± 20 nm). ROS and cell death are blocked either by inhibiting the mitochondrial electron transport chain or by mitochondria-specific antioxidants. These results show that mitochondria are an important source of toxic oxygen radicals in blue light-exposed RPE cells and may indicate new approaches for treating AMD using mitochondria-targeted antioxidants.
Journal of Biological …, 2012
Background: High levels of all-trans-retinal (atRAL) are associated with photoreceptor degeneration. Results: atRAL promotes NADPH oxidase-mediated overproduction of intracellular reactive oxygen species. Conclusion: A cascade of signaling events is demonstrated to underlie the action of atRAL in photoreceptor degeneration in mice. Significance: Mechanistic elucidation of atRAL-mediated photoreceptor degeneration is essential for understanding the molecular pathogenesis of Stargardt disease and other types of retinal degeneration. Compromised clearance of all-trans-retinal (atRAL), a component of the retinoid cycle, increases the susceptibility of mouse retina to acute light-induced photoreceptor degeneration. Abca4 ؊/؊ Rdh8 ؊/؊ mice featuring defective atRAL clearance were used to examine the one or more underlying molecular mechanisms, because exposure to intense light causes severe photoreceptor degeneration in these animals. Here we report that bright light exposure of Abca4 ؊/؊ Rdh8 ؊/؊ mice increased atRAL levels in the retina that induced rapid NADPH oxidasemediated overproduction of intracellular reactive oxygen species (ROS). Moreover, such ROS generation was inhibited by blocking phospholipase C and inositol 1,4,5-trisphosphate-induced Ca 2؉ release, indicating that activation occurs upstream of NADPH oxidase-mediated ROS generation. Because multiple upstream G protein-coupled receptors can activate phospholipase C, we then tested the effects of antagonists of serotonin 2A (5-HT 2A R) and M 3-muscarinic (M 3 R) receptors and found they both protected Abca4 ؊/؊ Rdh8 ؊/؊ mouse retinas from light-induced degeneration. Thus, a cascade of signaling events appears to mediate the toxicity of atRAL in light-induced photoreceptor degeneration of Abca4 ؊/؊ Rdh8 ؊/؊ mice. A similar mechanism may be operative in human Stargardt disease and age-related macular degeneration. To sustain vision, all-trans-retinal (atRAL), 2 released from light-activated visual pigments, including rhodopsin, must be