title: Antithrombin effects on endotoxin-induced microcirculatory disorders are mediated mainly by its interaction with microvascular endothelium.
abstract: OBJECTIVE
  To investigate whether the protective effect of antithrombin III, which has been shown to exert beneficial effects during septic disorders, including reduction of endotoxin-associated leukocyte/endothelial cell interaction and capillary perfusion failure, is mainly based on its anticoagulant capacity or direct effects on the microvascular endothelium.
  
  
  METHODS
  Animal study with three treatment groups.
  
  
  METHODS
  Animal research facility.
  
  
  METHODS
  Syrian golden hamsters, 6-8 wks old with a body weight of 60-80 g.
  
  
  METHODS
  In skinfold preparations of hamsters, normotensive endotoxemia was induced by intravenous administration of 2 mg/kg endotoxin (lipopolysaccharide, 2 mg/kg). Antithrombin III (n = 7 animals; 250 units/kg) or tryptophan49-blocked antithrombin III (n = 6; 250 units/kg) was substituted intravenously 5 mins before lipopolysaccharide administration. Saline-treated animals (n = 11), receiving only lipopolysaccharide, served as controls. Tryptophan49-blocked antithrombin III binds to glycosaminoglycans at the endothelial surface to a significantly lower extent while retaining its progressive anticoagulant effects.
  
  
  RESULTS
  Compared with controls, antithrombin III significantly reduced lipopolysaccharide-induced arteriolar and venular leukocyte adherence (p < .01) and prevented depression of functional capillary density (p < .01), whereas tryptophan49-blocked antithrombin III failed to significantly improve both variables. As measured in vivo by a monoclonal fluorescein isothiocyanate-labeled anti-antithrombin III antibody and intravital microscopy, the lack of effect of tryptophan49-blocked antithrombin III was associated with significantly lower antithrombin III/endothelium binding coefficients after 1 hr, 3 hrs, and 24 hrs of endotoxemia (p < .01).
  
  
  CONCLUSIONS
  We conclude that specific antithrombin III interactions with cell-surface glycosaminoglycans on the endothelium rather than anticoagulant properties are the mechanism of antithrombin III-mediated attenuation of leukocyte/endothelial cell interaction and capillary perfusion failure.
authors.affiliation: Department of Surgery, Klinikum Grosshadern, Ludwig-Maximilians University, Munich, Germany.
authors.forename: Johannes N
authors.initials: JN
authors.lastname: Hoffmann
authors.forename: Brigitte
authors.initials: B
authors.lastname: Vollmar
authors.forename: Jürgen
authors.initials: J
authors.lastname: Römisch
authors.forename: Dietrich
authors.initials: D
authors.lastname: Inthorn
authors.forename: Friedrich W
authors.initials: FW
authors.lastname: Schildberg
authors.forename: Michael D
authors.initials: MD
authors.lastname: Menger
mesh_terms: D000818:Animals; D000990:Antithrombin III / Q000378:metabolism / Q000494:pharmacology*; D001777:Blood Coagulation / Q000187:drug effects / Q000502:physiology; D006224:Cricetinae; D004730:Endothelium, Vascular / Q000502:physiology*; D004731:Endotoxins / Q000494:pharmacology*; D006025:Glycosaminoglycans / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D008647:Mesocricetus; D008833:Microcirculation / Q000502:physiology*; D008856:Microscopy, Fluorescence; D014364:Tryptophan / Q000494:pharmacology
chemical_list: D004731:Endotoxins; D006025:Glycosaminoglycans; D008070:Lipopolysaccharides; D014364:Tryptophan; D000990:Antithrombin III