Redox active hemoglobin enhances lipopolysaccharide-induced injury to cultured bovine endothelial cells.
The interaction of cell-free hemoglobin with lipopolysaccharide (LPS) is thought to aggravate the pathophysiology of sepsis and/or septic shock. This study examines the possible modulatory role of cell-free hemoglobin on LPS-induced apoptosis of cultured bovine aortic endothelial cells. Experiments were performed with or without fetal bovine serum, a source of LPS-binding protein and soluble CD14. In the absence of serum, LPS alone or coincubated with purified bovine hemoglobin (BvHb), human hemoglobin (Hb), or alpha-cross-linked Hb (alphaalphaHb) did not induce apoptosis. In the presence of serum, LPS induced significant apoptosis. LPS combined with BvHb, Hb, or alphaalphaHb produced the same extent of apoptosis as LPS alone. To examine whether the H(2)O(2)-driven redox activity of hemoglobin alters LPS-induced apoptosis, glucose oxidase was added to the system to generate a subtoxic flux of H(2)O(2). The combined treatment of LPS, glucose oxidase, and BvHb, Hb, or alphaalphaHb enhanced apoptosis compared with LPS alone. These findings support a possible mechanism whereby the redox cycling of hemoglobin, and not its direct interaction with LPS, contributes to the hemoglobin-mediated enhancement of LPS-related pathophysiology.
D'Agnillo
Felice
F
Center for Biologics Evaluation and Research, Food and Drug Administration, 29 Lincoln Drive, Bldg. 29, Rm. 129, Bethesda, MD 20892, USA. dagnillo@cber.fda.gov
D000818:Animals; D001011:Aorta / Q000166:cytology; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D001798:Blood Proteins / Q000494:pharmacology; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002417:Cattle; D002453:Cell Cycle / Q000502:physiology; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D006454:Hemoglobins / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000633:toxicity*; D016877:Oxidants / Q000494:pharmacology; D010084:Oxidation-Reduction
D001798:Blood Proteins; D006454:Hemoglobins; D008070:Lipopolysaccharides; D016877:Oxidants; D006861:Hydrogen Peroxide; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases