Dragana Škiljić1,2, Anne Petersen1, Jan-Olof Karlsson3, Anders Behndig4, Staffan Nilsson5, Madeleine Zetterberg1,2. 1. a Department of Clinical Neuroscience/Ophthalmology, Institute of Neuroscience and Physiology , The Sahlgrenska Academy at University of Gothenburg , Gothenburg , Sweden. 2. b Department of Ophthalmology , Sahlgrenska University Hospital , Mölndal , Sweden. 3. c Department of Medical Chemistry and Cell Biology , Institute of Biomedicine, The Sahlgrenska Academy at University of Gothenburg , Gothenburg , Sweden. 4. d Department of Clinical Sciences/Ophthalmology , Umeå University , Umeå , Sweden. 5. e Department of Mathematical Statistics, Institute of Mathematical Sciences , Chalmers University of Technology , Gothenburg , Sweden.
Abstract
PURPOSE: Protective effects of estradiol against H2O2-induced oxidative stress have been demonstrated in lens epithelial cells. The purpose of this study was to investigate the effects of 17β-estradiol (E2) on the different superoxide dismutase (SOD) isoenzymes, SOD-1, SOD-2, and SOD-3, as well as estrogen receptors (ERs), ERα and ERβ, in primary cultured human lens epithelial cells (HLECs). MATERIALS AND METHODS: HLECs were exposed to 0.1 µM or 1 µM E2 for 1.5 h and 24 h after which the effects were studied. Protein expression and immunolocalization of SOD-1, SOD-2, ERα, and ERβ were studied with Western blot and immunocytochemistry. Total SOD activity was measured, and gene expression analyses were performed for SOD1, SOD2, and SOD3. RESULTS: Increased SOD activity was seen after 1.5 h exposure to both 0.1 µM and 1 µM E2. There were no significant changes in protein or gene expression of the different SODs. Immunolabeling of SOD-1 was evident in the cytosol and nucleus; whereas, SOD-2 was localized in the mitochondria. Both ERα and ERβ were immunolocalized to the nucleus, and mitochondrial localization of ERβ was evident by colocalization with MitoTracker. Both ERα and ERβ showed altered protein expression levels after exposure to E2. CONCLUSIONS: The observed increase in SOD activity after exposure to E2 without accompanying increase in gene or protein expression supports a role for E2 in protection against oxidative stress mediated through non-genomic mechanisms.
PURPOSE: Protective effects of estradiol against H2O2-induced oxidative stress have been demonstrated in lens epithelial cells. The purpose of this study was to investigate the effects of 17β-estradiol (E2) on the different superoxide dismutase (SOD) isoenzymes, SOD-1, SOD-2, and SOD-3, as well as estrogen receptors (ERs), ERα and ERβ, in primary cultured human lens epithelial cells (HLECs). MATERIALS AND METHODS: HLECs were exposed to 0.1 µM or 1 µM E2 for 1.5 h and 24 h after which the effects were studied. Protein expression and immunolocalization of SOD-1, SOD-2, ERα, and ERβ were studied with Western blot and immunocytochemistry. Total SOD activity was measured, and gene expression analyses were performed for SOD1, SOD2, and SOD3. RESULTS: Increased SOD activity was seen after 1.5 h exposure to both 0.1 µM and 1 µM E2. There were no significant changes in protein or gene expression of the different SODs. Immunolabeling of SOD-1 was evident in the cytosol and nucleus; whereas, SOD-2 was localized in the mitochondria. Both ERα and ERβ were immunolocalized to the nucleus, and mitochondrial localization of ERβ was evident by colocalization with MitoTracker. Both ERα and ERβ showed altered protein expression levels after exposure to E2. CONCLUSIONS: The observed increase in SOD activity after exposure to E2 without accompanying increase in gene or protein expression supports a role for E2 in protection against oxidative stress mediated through non-genomic mechanisms.
Authors: Carlos A Silva-Islas; María E Chánez-Cárdenas; Diana Barrera-Oviedo; Alma Ortiz-Plata; José Pedraza-Chaverri; Perla D Maldonado Journal: Antioxidants (Basel) Date: 2019-09-18