Role for iron in reactive oxygen species-mediated cytotoxicity to cultured rat gastric mucosal cells.
The gastric epithelium is exposed to oxygen species that are generated within the lumen. Reactive oxygen species, enzymatically generated, cause injury to cultured rat gastric mucosal cells. Much interest has been focused on the role of iron in producing oxidant-mediated injury to the gastric mucosa, because iron is a catalyst that promotes the production of .OH possibly from O2-. and H2O2 (Haber-Weiss reaction) or from H2O2 alone (Fenton reaction). With the use of an iron chelator and an iron binding protein, we examined the role of iron in producing oxidant-mediated injury to cultured gastric mucosal cells. Reactive oxygen species and H2O2 were generated by hypoxanthine-xanthine oxidase and glucose-glucose oxidase, respectively, in buffer without iron. Pretreatment with deferoxamine diminished hypoxanthine-xanthine oxidase-induced 51Cr release from prelabeled cells, dose dependently. Furthermore, addition of deferoxamine to the reactive oxygen species-generating system also protected against the injury. However, apotransferrin (which binds extracellular iron) failed to protect cells. Pretreatment with .OH scavengers was partially protective. Depletion of glutathione with diethyl maleate enhanced reactive oxygen species-mediated cytolysis; such cytolysis was inhibited by deferoxamine. Deferoxamine also decreased 51Cr release induced by glucose-glucose oxidase. We conclude that intracellular iron plays a crucial role in mediating oxygen radical damage to gastric mucosal cells. The .OH, produced from H2O2 by the iron-catalyzed Fenton reaction, seems to be the main mediator of oxidant-induced cytotoxicity to gastric mucosal cells in vitro.
Hiraishi
H
H
Department of Medicine, Veterans Affairs Medical Center, Long Beach 90822.
Terano
A
A
Ota
S
S
Mutoh
H
H
Razandi
M
M
Sugimoto
T
T
Ivey
K J
KJ
D000818:Animals; D001059:Apoproteins / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003676:Deferoxamine / Q000494:pharmacology*; D005260:Female; D016166:Free Radical Scavengers; D005609:Free Radicals; D005753:Gastric Mucosa / Q000166:cytology* / Q000187:drug effects / Q000502:physiology; D005978:Glutathione / Q000378:metabolism; D006878:Hydroxides / Q000378:metabolism; D017665:Hydroxyl Radical; D007501:Iron / Q000502:physiology*; D007700:Kinetics; D008297:Male; D008298:Maleates / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D013481:Superoxides / Q000378:metabolism; D014168:Transferrin / Q000494:pharmacology*
D001059:Apoproteins; D016166:Free Radical Scavengers; D005609:Free Radicals; D006878:Hydroxides; D008298:Maleates; D014168:Transferrin; C028284:apotransferrin; D013481:Superoxides; D017665:Hydroxyl Radical; D007501:Iron; C014476:diethyl maleate; D005978:Glutathione; D003676:Deferoxamine