ABSTRACT
PURPOSE: The cytotoxic effects of oxidative stress, which play an important role in ocular diseases, are well known. In this study, we investigated the effect of non-lethal doses of oxidative stress on various cell functions, namely cell viability, cell attachment and cell migration in a widely used retinal pigment epithelium (RPE) cell line (ARPE-19). METHODS: A single exposure to various concentrations of hydrogen peroxide (H(2)O(2)) was used to establish a dose response for H(2)O(2)-induced cell death. Other cellular responses, such as changes in cell attachment and migration, were monitored after exposure to increasing doses. Finally, the effects of preconditioning cells with increasing non-lethal doses of H(2)O(2), with and without a subsequent exposure to lethal doses of H(2)O(2), were determined. RESULTS: The optimum dose for inducing cell death in ARPE-19 cells was between 900 and 1000 microm H(2)O(2). Preconditioning the cells with 1, 10 and 50 microm of H(2)O(2) provided a dose-dependent protection against cell death induced by a lethal dose (900-1000 microm) of H(2)O(2). Preconditioning with higher doses caused cells to become more susceptible to the cytotoxic effects of the lethal dose. Although H(2)O(2) increased cell attachment in lower doses, it induced a dose-dependent inhibition of cell attachment to the substrate in higher doses. H(2)O(2) did not affect cell migration in sub-lethal doses. CONCLUSION: Preconditioning RPE cells with limited exposure to non-lethal oxidative stress confers significant protection against subsequent H(2)O(2)-induced cell death. It also affects cell attachment in a dose-specific manner. This finding may help in understanding the pathogenesis of diseases in which oxidative stress plays an important role and in determining the suitability of certain treatment strategies, in particular RPE transplantation in the treatment of age-related macular degeneration.
