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Cannabinoid receptor 1 blockade protects human retinal pigment epithelial cells from oxidative injury

BACKGROUND: Because oxidative stress is assumed to be a key mechanism in the pathological process of age-related macular degeneration (AMD), increasing numbers of studies have focused on discovering new pathways and treatments for reducing oxidative damage. Our work investigates the potential role o...

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Autores principales: Wei, Yan, Wang, Xu, Zhao, Feng, Zhao, Pei-quan, Kang, Xiao-li
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Molecular Vision 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3580988/
https://www.ncbi.nlm.nih.gov/pubmed/23441106
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author Wei, Yan
Wang, Xu
Zhao, Feng
Zhao, Pei-quan
Kang, Xiao-li
author_facet Wei, Yan
Wang, Xu
Zhao, Feng
Zhao, Pei-quan
Kang, Xiao-li
author_sort Wei, Yan
collection PubMed
description BACKGROUND: Because oxidative stress is assumed to be a key mechanism in the pathological process of age-related macular degeneration (AMD), increasing numbers of studies have focused on discovering new pathways and treatments for reducing oxidative damage. Our work investigates the potential role of the cannabinoid receptor 1 (CB1) in oxidative stress of primary human retinal pigment epithelial (RPE) cells, a cellular model of AMD. METHODS: Primary human RPE cells were cultured and exposed to hydrogen peroxide for 24 h to induce oxidative damage. The expression of and changes in the CB1 receptor were determined with western blot assay and confocal imaging. The CB1 receptor in the RPE cells was inhibited with small interfering RNA (siRNA) or rimonabant (SR141716). Cell viability, apoptosis, and reactive oxygen species production were measured by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and sulforhodamine B assay, annexin V and propidium iodide staining, and the dichlorofluorescein fluorescence assay, respectively. Intracellular superoxide dismutase activity was assayed with a commercially available assay kit. Phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) protein expression and activation of signaling molecules were assessed with western blot analysis. RESULTS: We showed that human RPE cells express the CB1 receptor. In addition, oxidative stress upregulates the expression of the CB1 receptor. Deleting the CB1 receptor or treating with the CB1 receptor antagonist rimonabant (SR141716) rescued RPE cells from hydrogen peroxide–induced oxidative damage. Rimonabant pretreatment effectively reduced the apoptosis of RPE cells, inhibited the generation of intracellular reactive oxygen species and elevated the activity of superoxide dismutase. In addition, rimonabant significantly strengthened the oxidative stress-induced activation of the PI3K/Akt signaling pathway. CONCLUSIONS: The results demonstrate the expression and regulation of CB1 receptors in human RPE cells. Inhibiting the CB1 receptor may be an effective therapeutic strategy for AMD by downregulating oxidative stress signaling and facilitating PI3K/Akt activation.
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spelling pubmed-35809882013-02-25 Cannabinoid receptor 1 blockade protects human retinal pigment epithelial cells from oxidative injury Wei, Yan Wang, Xu Zhao, Feng Zhao, Pei-quan Kang, Xiao-li Mol Vis Research Article BACKGROUND: Because oxidative stress is assumed to be a key mechanism in the pathological process of age-related macular degeneration (AMD), increasing numbers of studies have focused on discovering new pathways and treatments for reducing oxidative damage. Our work investigates the potential role of the cannabinoid receptor 1 (CB1) in oxidative stress of primary human retinal pigment epithelial (RPE) cells, a cellular model of AMD. METHODS: Primary human RPE cells were cultured and exposed to hydrogen peroxide for 24 h to induce oxidative damage. The expression of and changes in the CB1 receptor were determined with western blot assay and confocal imaging. The CB1 receptor in the RPE cells was inhibited with small interfering RNA (siRNA) or rimonabant (SR141716). Cell viability, apoptosis, and reactive oxygen species production were measured by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and sulforhodamine B assay, annexin V and propidium iodide staining, and the dichlorofluorescein fluorescence assay, respectively. Intracellular superoxide dismutase activity was assayed with a commercially available assay kit. Phosphoinositide 3-kinase/protein kinase B (PI3K/Akt) protein expression and activation of signaling molecules were assessed with western blot analysis. RESULTS: We showed that human RPE cells express the CB1 receptor. In addition, oxidative stress upregulates the expression of the CB1 receptor. Deleting the CB1 receptor or treating with the CB1 receptor antagonist rimonabant (SR141716) rescued RPE cells from hydrogen peroxide–induced oxidative damage. Rimonabant pretreatment effectively reduced the apoptosis of RPE cells, inhibited the generation of intracellular reactive oxygen species and elevated the activity of superoxide dismutase. In addition, rimonabant significantly strengthened the oxidative stress-induced activation of the PI3K/Akt signaling pathway. CONCLUSIONS: The results demonstrate the expression and regulation of CB1 receptors in human RPE cells. Inhibiting the CB1 receptor may be an effective therapeutic strategy for AMD by downregulating oxidative stress signaling and facilitating PI3K/Akt activation. Molecular Vision 2013-02-13 /pmc/articles/PMC3580988/ /pubmed/23441106 Text en Copyright © 2013 Molecular Vision. http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Wei, Yan
Wang, Xu
Zhao, Feng
Zhao, Pei-quan
Kang, Xiao-li
Cannabinoid receptor 1 blockade protects human retinal pigment epithelial cells from oxidative injury
title Cannabinoid receptor 1 blockade protects human retinal pigment epithelial cells from oxidative injury
title_full Cannabinoid receptor 1 blockade protects human retinal pigment epithelial cells from oxidative injury
title_fullStr Cannabinoid receptor 1 blockade protects human retinal pigment epithelial cells from oxidative injury
title_full_unstemmed Cannabinoid receptor 1 blockade protects human retinal pigment epithelial cells from oxidative injury
title_short Cannabinoid receptor 1 blockade protects human retinal pigment epithelial cells from oxidative injury
title_sort cannabinoid receptor 1 blockade protects human retinal pigment epithelial cells from oxidative injury
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3580988/
https://www.ncbi.nlm.nih.gov/pubmed/23441106
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