title: Interactions of hemoglobin with hydrogen peroxide alters thiol levels and course of endothelial cell death.
abstract: We investigated cellular injury and death induced by ultrapure human Hb (HbA(0)) and its diaspirin cross-linked derivative DBBF-Hb in normal and glutathione (GSH)-depleted bovine aortic endothelial cells subjected to hydrogen peroxide (H(2)O(2)). HbA(0) underwent extensive degradation and heme loss, whereas DBBF-Hb persisted longer in its ferryl (Fe(4+)) form. The formation of ferryl HbA(0) or ferryl DBBF-Hb was associated with a significant decrease in endothelial cell GSH compared with the addition of H(2)O(2) or Hbs alone. This effect was inhibited by catalase, but not by superoxide dismutase or deferoxamine mesylate. The presence of HbA(0) and DBBF-Hb reduced H(2)O(2)-induced apoptosis, as measured by cell morphology, annexin V binding assay, and caspase inhibition, consistent with the ability to consume H(2)O(2) in an enzyme-like fashion. However, the pattern of cell death and injury produced by HbA(0) and DBBF-Hb appeared to be distinctly different among proteins as well as among cells with and without GSH. These findings may have important implications for the use of cell-free Hb as oxygen therapeutics in patients with coexisting pathologies who may lack antioxidant protective mechanisms.
authors.affiliation: Laboratory of Plasma Derivatives, Division of Hematology, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892, USA.
authors.forename: F
authors.initials: F
authors.lastname: D'Agnillo
authors.forename: A I
authors.initials: AI
authors.lastname: Alayash
mesh_terms: D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D001011:Aorta / Q000166:cytology / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D017209:Apoptosis / Q000187:drug effects; D001241:Aspirin / Q000031:analogs & derivatives* / Q000494:pharmacology; D002417:Cattle; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D005978:Glutathione / Q000378:metabolism; D006418:Heme / Q000378:metabolism; D006441:Hemoglobin A / Q000494:pharmacology*; D006454:Hemoglobins / Q000494:pharmacology; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D016877:Oxidants / Q000494:pharmacology*; D013438:Sulfhydryl Compounds / Q000378:metabolism*
chemical_list: D017304:Annexin A5; D006454:Hemoglobins; D016877:Oxidants; D013438:Sulfhydryl Compounds; C115379:bis(3,5-dibromosalicyl)sebacate-crosslinked hemoglobin; D006418:Heme; C028881:hemoglobin A(0); D006441:Hemoglobin A; D006861:Hydrogen Peroxide; D005978:Glutathione; D001241:Aspirin