Effects of 17-Estradiol on Activity, Gene and Protein Expression of Superoxide Dismutases in Primary Cultured Human Lens Epithelial Cells

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 17-estradiol (E2) on the different superoxide dismutase (SOD) isoenzymes, SOD-1, SOD-2, and SOD-3, as well as estrogen receptors (ERs), ER and ER, in primary cultured human lens epithelial cells (HLECs).Materials and methods: HLECs were exposed to 0.1 mu M or 1 mu M E2 for 1.5h and 24h after which the effects were studied. Protein expression and immunolocalization of SOD-1, SOD-2, ER, and ER 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 1.5h exposure to both 0.1 mu M and 1 mu M 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 ER and ER were immunolocalized to the nucleus, and mitochondrial localization of ER was evident by colocalization with MitoTracker. Both ER and ER 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.