Role of iron and superoxide in mediating hydrogen peroxide injury to cultured rat gastric cells.

Abstract

Gastric epithelium is exposed to toxic, reactive oxygen species generated within the lumen. The present study examined the role of cellular iron and superoxide (O2-) in mediating hydrogen peroxide (H2O2)-induced damage to cultured gastric mucosal cells. H2O2 was generated by glucose oxidase acting on b-D(+)glucose. Cytotoxicity was assessed by 51Cr release from prelabeled cells. Deferoxamine (a chelator of Fe3+) prevented injury induced by H2O2, whether present before or during H2O2 production. In contrast, whereas the presence of phenanthroline (a chelator of Fe2+) during the cytotoxicity assay prevented damage, prior treatment with the agent did not; this suggested that cellular Fe3+ reduced to Fe2+ upon exposure to H2O2 is responsible for damage. Neither extracellular superoxide dismutase nor inhibitors of xanthine oxidase (a possible source of cellular O2- production) protected against H2O2. Further, protection by iron chelators was not associated with modulation of endogenous antioxidants. Deferoxamine and phenanthroline protect cells from H2O2 by chelating stored iron (Fe3+) or reduced iron (Fe2+), respectively. Reduction of cellular Fe3+ appears to be a prerequisite for mediation of damage, but this reduction is independent of extracellular O2- or cellular xanthine oxidase-derived O2-.