1. Effects of 17β-Estradiol on Activity, Gene and Protein Expression of Superoxide Dismutases in Primary Cultured Human Lens Epithelial Cells
- Author
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Madeleine Zetterberg, Anders Behndig, A. Petersen, Dragana Škiljić, Staffan Nilsson, and Jan Olof Karlsson
- Subjects
0301 basic medicine ,Cell- och molekylärbiologi ,Blotting, Western ,Real-Time Polymerase Chain Reaction ,medicine.disease_cause ,Gene Expression Regulation, Enzymologic ,Superoxide dismutase ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,0302 clinical medicine ,Lens, Crystalline ,medicine ,oxidative stress ,Humans ,Gene and protein expression ,Cells, Cultured ,17-estradiol ,Cell Nucleus ,lens epithelial cells ,Estradiol ,biology ,Superoxide Dismutase ,Chemistry ,Estrogen Receptor alpha ,Nuclear Proteins ,RNA-Binding Proteins ,Epithelial Cells ,Estrogens ,superoxide dismutase ,Immunohistochemistry ,Sensory Systems ,Mitochondria ,Cell biology ,Isoenzymes ,Oxidative Stress ,Ophthalmology ,Cell and molecular biology ,030104 developmental biology ,medicine.anatomical_structure ,cataract ,Lens (anatomy) ,030221 ophthalmology & optometry ,biology.protein ,Carrier Proteins ,Cell and Molecular Biology ,Oxidative stress - 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.
- Published
- 2018