Premature induction of aging in sublethally H 2O 2-treated young MRC5 fibroblasts correlates with increased glutathione peroxidase levels and resistance to DNA breakage

Abstract

Human MRC5 fibroblasts, at different passages in cultures, were used as an in vitro model to assess variations and/or induction of aging parameters under basal conditions or following sublethal oxidative stress by H 2O 2. DNA sensitivities to oxidatively-induced breakage, rather than basal levels of damaged DNA, were significantly different between cultures at low and high population doubling level (PDL): old cells maintained most of their DNA integrity even at high concentrations of H 2O 2, while young cells showed more extensive DNA damage which developed in a dose-dependent fashion. However, young cells pretreated with low doses of H 2O 2 exhibited increased resistance against further oxidative damage to DNA thus reproducing a senescent-like profile of sensitivity. In turn, DNA from old cultures incubated in a NAD precursor-free medium was more prone to H 2O 2-induced strand breaks mimicking DNA sensitivity of young cells. The extent of oxidatively-induced DNA damage in MRC5 populations correlated inversely with the levels of glutathione peroxidase (GPx) activity that almost doubled when cells passed from the young to the senescent stage. In addition, H 2O 2-pretreatment of young cells induced an increase in GPx expression approaching old cell values and promoted also the premature appearance of neutral β-galactosidase activity and decreased c- fos expression upon serum stimulation, both of which were assumed to be characteristic traits of the senescent phenotype.