title: Biological response of human aortic endothelial cells exposed to acellular hemoglobin solutions developed as potential blood substitutes.
abstract: The cardiovascular effects of hemoglobin-based oxygen carriers (HBOCs) are mainly related to their nitric oxide (NO) scavenging properties but other effects such as the impact of these hemoglobins on the endothelial cell (EC) biology are not well understood. We hypothesized that HBOCs could modify EC functions by altering gene expression, in particular the endothelial NO synthase (NOS3) and/or by activating EC. Cultured human aortic endothelial cells (HAEC) were incubated for 3 hours with purified cell-free Hb, Dex-BTC-Hb or alpha alpha-Hb (16 g/L). Expression of NOS3 mRNA and protein were assessed by semi-quantitative RT-PCR and Western blot respectively immediately after and 24 hours after incubation. The expression and localization of the adhesion molecule ICAM-1 were detected by fluorescence microscopy. None of the solutions tested modified NOS3 mRNA and protein expression despite adequate controls that up- or down-regulate NOS3 expression. The expression and the localization of ICAM-1 on the cell membrane were modified after 3 hours of incubation with all the hemoglobin solutions tested in a manner similar to tumor necrosis factor-alpha. In conclusion, HAEC incubation with clinically relevant concentrations of HBOCs induced changes in the pattern of ICAM-1 expression consistent with cell activation/cell signaling mechanisms. However, HBOCs did not alter NOS3 gene expression.
authors.affiliation: Laboratoire d'Hématologie-Physiologie, UPRES EA 3452, Faculté de Pharmacie, 54000 Nancy, France. marie.toussaint@pharma.uhp-nancy.fr
authors.forename: M
authors.initials: M
authors.lastname: Toussaint-Hacquard
authors.forename: Y
authors.initials: Y
authors.lastname: Devaux
authors.forename: D
authors.initials: D
authors.lastname: Longrois
authors.forename: B
authors.initials: B
authors.lastname: Faivre-Fiorina
authors.forename: S
authors.initials: S
authors.lastname: Muller
authors.forename: J F
authors.initials: JF
authors.lastname: Stoltz
authors.forename: C
authors.initials: C
authors.lastname: Vigneron
authors.forename: P
authors.initials: P
authors.lastname: Menu
mesh_terms: D001011:Aorta / Q000166:cytology / Q000187:drug effects; D001802:Blood Substitutes / Q000737:chemistry / Q000494:pharmacology*; D015153:Blotting, Western; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003720:Densitometry; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006454:Hemoglobins / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D066298:In Vitro Techniques; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis / Q000378:metabolism; D019001:Nitric Oxide Synthase / Q000096:biosynthesis / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III; D012313:RNA / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D001802:Blood Substitutes; D006454:Hemoglobins; D012333:RNA, Messenger; D018799:Intercellular Adhesion Molecule-1; D012313:RNA; C496314:NOS3 protein, human; D019001:Nitric Oxide Synthase; D052250:Nitric Oxide Synthase Type III