Purpose: The purpose of this study was to examine the effects of 17β-estradiol on proliferation, cell death and redox status in cultured human lens epithelial cells (HLECs).
Methods: HLECs were exposed to 17β-estradiol after which cell viability was measured by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) and the number of mitotic and apoptotic cell nuclei was determined after staining with Hoechst 33342. Apoptosis was also determined by measuring caspase-3 activity and propidium iodide was used to determine the proportion of non-viable cells. Pro- and antioxidative effects of 17β-estradiol was investigated by measuring peroxides, superoxides and glutathione, using dichlorofluorescein diacetate (DCFH-DA), dihydroethidium (HET), and monochlorobimane (MCB), respectively. Effects on mitochondrial membrane potential were determined using 5,5',6,6'-tetrachloro-1,1',3,3'- tetraethylbenzimidazolylcarbocyanine iodide (JC-1). The ability of 17β-estradiol to prevent reactive oxygen species (ROS)-production in HLECs after exposure to 25 µM H₂O₂ for 24h was also measured.
Results: This study demonstrates increased mitotic activity in HLECs exposed to physiologic concentrations of 17β-estradiol (1 nM). Pharmacological concentrations of 17β-estradiol caused increased number of apoptotic cell nuclei and caspase-3 activation. Physiologic concentrations of 17β-estradiol (0.1-10 nM) stabilized the mitochondrial membrane potential. Similar or slightly higher concentrations of 17β-estradiol (0.01-1 µM) protected against H₂O₂-induced oxidative stress as evident by decreased levels of peroxides and superoxides.
Conclusions: The present study demonstrates mitogenic and anti-oxidative effects of 17β-estradiol at physiologic concentrations, whereas pharmacological levels induced oxidative stress and acted pro-apoptotic in cultured lens cells.