Preconditioning protects the retinal pigment epithelium cells from oxidative stress‐induced cell death

Abstract

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 lm H 2 O 2 . Preconditioning the cells with 1, 10 and 50 lm of H 2 O 2 provided a dose-dependent protection against cell death induced by a lethal dose (900-1000 lm) 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.

Identification of mammary epithelial cells subject to chronic oxidative stress in mammary epithelium of young women and teenagers living in USA

“…Formalin-fixed, paraffin-embedded (FFPE) tissues (n = 58, ages [14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][29][30] were sectioned at 5 mm, transferred onto Fisher Superfrost Plus slides, deparaffinized, rehydrated and then subjected to antigen retrieval using Vector Antigen Unmasking Solution, pH 6.0 (Vector Laboratories) for 1 h at 80°C. The sections were incubated in a humidity chamber overnight at 4°C with a mouse monoclonal antibody developed against 4HNE-modified low-density lipoprotein (clone NA59) that binds to lysine adducts of 4HNE.…”

“…[18][19][20][21][22][23][24][25][26][27] Of special relevance to carcinogenesis are adaptive responses to chronic oxidative stress identified that might lead to the evolution of subpopulations of cells that can evade mechanisms that limit the propagation of cells with oxidatively-damaged DNA, notably, increased resistance to apoptosis [28][29][30][31] and tolerance to DNA damage. [32][33][34] The chronic oxidative stress final-common-path hypothesis, referred to above, is based on the following considerations.…”

Advances in Regenerative Medicine

“…The majority of studies have experimented with hypoxia or anoxia alone, termed hypoxic preconditioning (HPC), while others include nutrient deprivation. In some studies, hydrogen peroxide (H 2 O 2 ) was used to simulate the ischaemic condition of oxidative stress (Li et al, 2009;Sharma et al, 2008). Furthermore, different HPC protocols, from the classical multiple cycles of brief hypoxia with intermittent reoxygenation to a single long-term exposure to hypoxia, have been employed to demonstrate the cytoprotective effect of HPC on stem and progenitor cells in vitro.…”

“…Preconditioning of model cells of retinal pigment epithelium with non-lethal oxidative stress protects these cells from cell death induced by oxidative-stress (Sharma et al, 2009). The cell line ARPE-19 was used to mimick the conditions of oxidative stress, which is encountered by retinal cells transplanted for repairing the age-related macular degeneration.…”

Cellular Stress Responses

“…There are examples of pre-treatments of cells to manipulate stress response pathways that minimise cellular damage and improve the transplantation outcome. Preconditioning of model cells of retinal pigment epithelium with non-lethal oxidative stress protects these cells from cell death induced by oxidative-stress (Sharma et al, 2009). The cell line ARPE-19 was used to mimick the conditions of oxidative stress, which is encountered by retinal cells transplanted for repairing the age-related macular degeneration.…”