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Crit Care Med. 2003 Apr;31(4):1147-53.

Gabexate mesilate, a synthetic anticoagulant, inhibits the expression of endothelial leukocyte adhesion molecules in vitro.

Author information

1
Department of Laboratory Medicine, Kumamoto University School of Medicine, Kumamoto, Japan.

Abstract

OBJECTIVE:

Gabexate mesilate, a synthetic protease inhibitor, has been shown to reduce endotoxin-induced pulmonary vascular injury in an animal model of sepsis by inhibiting leukocyte activation. We examined whether gabexate mesilate inhibits tumor necrosis factor-alpha-induced expression of leukocyte adhesion molecules in cultured endothelial cells.

DESIGN:

Prospective, randomized, controlled study.

SETTING:

Research laboratory at a university medical center.

SUBJECTS:

Cultured human umbilical vein endothelial cell (HUVECs).

INTERVENTIONS:

HUVECs were stimulated with tumor necrosis factor-alpha or lipopolysaccharide in the presence or absence of gabexate mesilate. Expression of E-selectin and intercellular adhesion molecule-1 was measured by cellular enzyme-linked immunosorbent assay. Messenger RNA levels of E-selectin and intercellular adhesion molecule-1 were determined by reverse transcription-polymerase chain reaction. DNA-binding activity of p65 in the nuclear extracts was evaluated by enzyme-linked immunosorbent assay. Nuclear translocation of nuclear factor-kappaB induced by tumor necrosis factor-alpha was evaluated by immunocytostaining and Western blot analysis. Degradation and phosphorylation of inhibitor of nuclear factor-kappaB (IkappaB) induced by tumor necrosis factor-alpha were evaluated by Western blot analysis.

MEASUREMENTS AND MAIN RESULTS:

Gabexate mesilate inhibited the tumor necrosis factor-alpha-induced increases in the endothelial expression of E-selectin and intercellular adhesion molecule-1 by inhibiting the transcription. Tumor necrosis factor-alpha-induced increase in DNA binding of p65 was inhibited by gabexate mesilate through inhibition of the nuclear translocation of p65. Gabexate mesilate inhibited the tumor necrosis factor-alpha-induced degradation of IkappaBalpha, an inhibitor of nuclear factor-kappaB, by inhibiting phosphorylation of IkappaBalpha in HUVECs.

CONCLUSIONS:

Gabexate mesilate inhibited the expression of leukocyte adhesion molecules by inhibiting the nuclear factor-kappaB-mediated transcription in HUVECs. Inhibition of nuclear factor-kappaB activation by gabexate mesilate could be explained by inhibition of degradation of IkappaB. Gabexate mesilate might reduce lipopolysaccharide-induced pulmonary vascular injury not only by inhibiting monocytic tumor necrosis factor-alpha production but by inhibiting the expression of endothelial leukocyte adhesion molecules.

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