Changes in Mitochondrial Morphology and Mitochondrial Fission/Fusion Gene Expression in Retinal Pigment Epithelial Cells During Oxidative Stress

Age-related macular degeneration (AMD) represents a serious impairment for the elderly. Because the pathogenesis of AMD has not been completely defined, the available therapeutic treatments are not ideal. Retinal pigment epithelial (RPE) cells are essential for photoreceptor cell maintenance and survival; however, the mechanisms underlying RPE cell damage and AMD remains to be elucidated. It is known that abnormal mitochondrial gene expression causes mitochondrial dysfunction, induces cell damage, and results in disease. In this study, ARPE-19 cells were treated with different concentrations of H2O2. It was found that excessive H2O2 concentration resulted in significant contraction of ARPE-19 cells and increased cell death, and destruction of mitochondrial structure as well as membrane and crest. RT-PCR results showed that decreased expression of the Fis1 gene was evident in H2O2-treated cells. There were no significant differences observed among the different H2O2 concentration groups. The expression of the fission genes, MTP18 and Dnmp1, and the fusion genes, Mnf1 and Mnf2, was not significantly different. Real-time PCR results revealed that the expression of the Fis1 gene decreased concomitantly with different concentrations of H2O2, whereas the expression of the Mfn2 gene increased by treatment with 200 μMH2O2. There were no significant differences in the expression of the other genes. These results indicate that abnormal expression of the mitochondrial Fis1 fission gene, and the Mfn2 fusion gene caused mitochondrial dysfunction in ARPE-19 cells. This indicates that the imbalance of mitochondrial dynamics may contribute to cell death in an oxidative stress environment.

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Sulforaphane Enhances the Ability of Human Retinal Pigment Epithelial Cell against Oxidative Stress, and Its Effect on Gene Expression Profile Evaluated by Microarray Analysis

Oxidative Medicine and Cellular Longevity ◽

10.1155/2013/413024 ◽

2013 ◽

Vol 2013 ◽

pp. 1-13 ◽

Cited By ~ 10

Author(s):

Liang Ye ◽

Ting Yu ◽

Yanqun Li ◽

Bingni Chen ◽

Jinshun Zhang ◽

...

Keyword(s):

Oxidative Stress ◽

Gene Expression ◽

Microarray Analysis ◽

Retinal Pigment Epithelial ◽

Nuclear Translocation ◽

Retinal Pigment ◽

Regulation Of Gene Expression ◽

Age Related Macular Degeneration ◽

Rpe Cells ◽

Pigment Epithelial

To gain further insights into the molecular basis of Sulforaphane (SF) mediated retinal pigment epithelial (RPE) 19 cell against oxidative stress, we investigated the effects of SF on the regulation of gene expression on a global scale and tested whether SF can endow RPE cells with the ability to resist apoptosis. The data revealed that after exposure to H2O2, RPE 19 cell viability was increased in the cells pretreated with SF compared to the cell not treated with SF. Microarray analysis revealed significant changes in the expression of 69 genes in RPE 19 cells after 6 hours of SF treatment. Based on the functional relevance, eight of the SF-responsive genes, that belong to antioxidant redox system, and inflammatory responsive factors were validated. The up-regulating translation of thioredoxin-1 (Trx1) and the nuclear translocation of Nuclear factor-like2 (Nrf2) were demonstrated by immunoblot analysis in SF treated RPE cells. Our data indicate that SF increases the ability of RPE 19 cell against oxidative stress through up-regulating antioxidative enzymes and down-regulating inflammatory mediators and chemokines. The results suggest that the antioxidant, SF, may be a valuable supplement for preventing and retarding the development of Age Related Macular Degeneration.

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Oxidative stress induces ferroptotic cell death in retinal pigment epithelial cells

Experimental Eye Research ◽

10.1016/j.exer.2018.08.019 ◽

2019 ◽

Vol 181 ◽

pp. 316-324 ◽

Cited By ~ 17

Author(s):

Kiyohito Totsuka ◽

Takashi Ueta ◽

Takatoshi Uchida ◽

Murilo F. Roggia ◽

Suguru Nakagawa ◽

...

Keyword(s):

Oxidative Stress ◽

Cell Death ◽

Epithelial Cells ◽

Retinal Pigment Epithelial ◽

Retinal Pigment ◽

Retinal Pigment Epithelial Cells ◽

Pigment Epithelial ◽

Pigment Epithelial Cells

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S-Allylmercapro-N-Acetylcysteine Attenuates the Oxidation-Induced Lens Opacification and Retinal Pigment Epithelial Cell Death In Vitro

Antioxidants ◽

10.3390/antiox8010025 ◽

2019 ◽

Vol 8 (1) ◽

pp. 25 ◽

Cited By ~ 1

Author(s):

Naphtali Savion ◽

Samia Dahamshi ◽

Milana Morein ◽

Shlomo Kotev-Emeth

Keyword(s):

Oxidative Stress ◽

Cell Death ◽

Retinal Pigment Epithelial ◽

Retinal Pigment Epithelial Cell ◽

Retinal Pigment ◽

Lens Opacity ◽

Glutathione Level ◽

Rpe Cells ◽

Pigment Epithelial ◽

Lens Opacification

The capacity of S-Allylmercapto-N-acetylcysteine (ASSNAC) to protect human retinal pigment epithelial (RPE) cells (line ARPE-19) and porcine lenses from oxidative stress was studied. Confluent ARPE-19 cultures were incubated with ASSNAC or N-acetyl-cysteine (NAC) followed by exposure to oxidants and glutathione level and cell survival were determined. Porcine lenses were incubated with ASSNAC and then exposed to H2O2 followed by lens opacity measurement and determination of glutathione (reduced (GSH) and oxidized (GSSG)) in isolated lens adhering epithelial cells (lens capsule) and fiber cells consisting the lens cortex and nucleus (lens core). In ARPE-19 cultures, ASSNAC (0.2 mM; 24 h) increased glutathione level by 2–2.5-fold with significantly higher increase in GSH compared to NAC treated cultures. Similarly, ex-vivo exposure of lenses to ASSNAC (1 mM) significantly reduced the GSSG level and prevented H2O2 (0.5 mM)-induced lens opacification. These results demonstrate that ASSNAC up-regulates glutathione level in RPE cells and protects them from oxidative stress-induced cell death as well as protects lenses from oxidative stress-induced opacity. Further validation of these results in animal models may suggest a potential use for ASSNAC as a protective therapy in retinal degenerative diseases as well as in attenuation of oxidative stress-induced lens opacity.

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Gossypol Acetic Acid Prevents Oxidative Stress-Induced Retinal Pigment Epithelial Necrosis by Regulating the FoxO3/Sestrin2 Pathway

Molecular and Cellular Biology ◽

10.1128/mcb.00178-15 ◽

2015 ◽

Vol 35 (11) ◽

pp. 1952-1963 ◽

Cited By ~ 10

Author(s):

Jakub Hanus ◽

Hongmei Zhang ◽

David H. Chen ◽

Qinbo Zhou ◽

Peng Jin ◽

...

Keyword(s):

Oxidative Stress ◽

Cell Death ◽

Acetic Acid ◽

Retinal Pigment Epithelial ◽

Potent Inhibitor ◽

Retinal Pigment ◽

Pigment Epithelial ◽

Antioxidative Response ◽

Age Related ◽

Gossypol Acetic Acid

The late stage of dry age-related macular degeneration (AMD), or geographic atrophy (GA), is characterized by extensive retinal pigment epithelial (RPE) cell death, and a cure is not available currently. We have recently demonstrated that RPE cells die from necrosis in response to oxidative stress, providing a potential novel mechanism for RPE death in AMD. In this study, we screened U.S. Food and Drug Administration-approved natural compounds and identified gossypol acetic acid (GAA) as a potent inhibitor of oxidative stress-induced RPE cell death. GAA induces antioxidative response and inhibits accumulation of excessive reactive oxygen species in cells, through which it prevents the activation of intrinsic necrotic pathway in response to oxidative stress. Sestrin2 (SESN2) is found to mediate GAA function in antioxidative response and RPE survival upon oxidative stress. Moreover, Forkhead box O3 transcription factor (FoxO3) is further found to be required for GAA-mediated SESN2 expression and RPE survival. Mechanistically, GAA promotes FoxO3 nuclear translocation and binding to theSESN2enhancer, which in turn increases its transcriptional activity. Taken together, we have identified GAA as a potent inhibitor of oxidative stress-induced RPE necrosis by regulating the FoxO3/SESN2 pathway. This study may have significant implications in the therapeutics of age-related diseases, especially GA.

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PRMT1 and PRMT4 Regulate Oxidative Stress-Induced Retinal Pigment Epithelial Cell Damage in SIRT1-Dependent and SIRT1-Independent Manners

Oxidative Medicine and Cellular Longevity ◽

10.1155/2015/617919 ◽

2015 ◽

Vol 2015 ◽

pp. 1-9 ◽

Cited By ~ 16

Author(s):

Dong-Il Kim ◽

Min-Jung Park ◽

Joo-Hee Choi ◽

In-Seon Kim ◽

Ho-Jae Han ◽

...

Keyword(s):

Oxidative Stress ◽

Diabetic Retinopathy ◽

Retinal Pigment Epithelial ◽

Cell Damage ◽

Retinal Pigment ◽

Arginine Methylation ◽

Type I ◽

Dependent Manner ◽

Sirt1 Expression ◽

Pigment Epithelial

Oxidative stress-induced retinal pigment epithelial (RPE) cell damage is involved in the progression of diabetic retinopathy. Arginine methylation catalyzed by protein arginine methyltransferases (PRMTs) has emerged as an important histone modification involved in diverse diseases. Sirtuin (SIRT1) is a protein deacetylase implicated in the onset of metabolic diseases. Therefore, we examined the roles of type I PRMTs and their relationship with SIRT1 in human RPE cells under H2O2-induced oxidative stress. H2O2treatment increased PRMT1 and PRMT4 expression but decreased SIRT1 expression. Similar to H2O2treatment, PRMT1 or PRMT4 overexpression increased RPE cell damage. Moreover, the H2O2-induced RPE cell damage was attenuated by PRMT1 or PRMT4 knockdown and SIRT1 overexpression. In this study, we revealed that SIRT1 expression was regulated by PRMT1 but not by PRMT4. Finally, we found that PRMT1 and PRMT4 expression is increased in the RPE layer of streptozotocin-treated rats. Taken together, we demonstrated that oxidative stress induces apoptosis both via PRMT1 in a SIRT1-dependent manner and via PRMT4 in a SIRT1-independent manner. The inhibition of the expression of type I PRMTs, especially PRMT1 and PRMT4, and increased SIRT1 could be therapeutic approaches for diabetic retinopathy.

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Cytoprotective Effects of Natural Highly Bio-Available Vegetable Derivatives on Human-Derived Retinal Cells

Nutrients ◽

10.3390/nu12030879 ◽

2020 ◽

Vol 12 (3) ◽

pp. 879 ◽

Cited By ~ 1

Author(s):

Ingrid Munia ◽

Laurent Gafray ◽

Marie-Agnès Bringer ◽

Pablo Goldschmidt ◽

Lil Proukhnitzky ◽

...

Keyword(s):

Oxidative Stress ◽

Cell Death ◽

Metabolic Activity ◽

Retinal Pigment Epithelial ◽

Caspase 3 ◽

Retinal Pigment ◽

Retinal Pigment Epithelial Cells ◽

Pigment Epithelial ◽

Pre Treatment ◽

Pigment Epithelial Cells

Retinal pigment epithelial cells are crucial for retina maintenance, making their cytoprotection an excellent way to prevent or slow down retinal degeneration. In addition, oxidative stress, inflammation, apoptosis, neovascularization, and/or autophagy are key pathways involved in degenerative mechanisms. Therefore, here we studied the effects of curcumin, lutein, and/or resveratrol on human retinal pigment epithelial cells (ARPE-19). Cells were incubated with individual or combined agent(s) before induction of (a) H2O2-induced oxidative stress, (b) staurosporin-induced apoptosis, (c) CoCl2-induced hypoxia, or (d) a LED-autophagy perturbator. Metabolic activity, cellular survival, caspase 3/7 activity (casp3/7), cell morphology, VEGF levels, and autophagy process were assessed. H2O2 provoked a reduction in cell survival, whereas curcumin reduced metabolic activity which was not associated with cell death. Cell death induced by H2O2 was significantly reduced after pre-treatment with curcumin and lutein, but not resveratrol. Staurosporin increased caspase-3/7 activity (689%) and decreased cell survival by 32%. Curcumin or lutein protected cells from death induced by staurosporin. Curcumin, lutein, and resveratrol were ineffective on the increase of caspase 3/7 induced by staurosporin. Pre-treatment with curcumin or lutein prevented LED-induced blockage of autophagy flux. Basal-VEGF release was significantly reduced by lutein. Therefore, lutein and curcumin showed beneficial protective effects on human-derived retinal cells against several insults.

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