Inhibition of the endothelial cell activation by antithrombin in vitro

Mitsuhiro Uchiba; Kenji Okajima; Christoph Kaun; Johann Wojta; Bernd R. Binder

doi:10.1160/TH04-03-0139

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2004; 92(06): 1420-1427
DOI: 10.1160/TH04-03-0139

Endothelium and Vascular Development

Schattauer GmbH

Inhibition of the endothelial cell activation by antithrombin in vitro

Mitsuhiro Uchiba

¹Department of Diagnostic Medicine, Graduate School of Medical Sciences, Kumamoto University, Kumamoto, Japan

,

Kenji Okajima

¹Department of Diagnostic Medicine, Graduate School of Medical Sciences, Kumamoto University, Kumamoto, Japan

,

Christoph Kaun

²Department of Internal Medicine II, University of Vienna, Vienna, Austria

,

Johann Wojta

²Department of Internal Medicine II, University of Vienna, Vienna, Austria

,

Bernd R. Binder

³Department of Vascular Biology and Thrombosis Research, University of Vienna, Vienna, Austria

› Author Affiliations

Abstract

Summary

We examined whether antithrombin (AT) inhibits tumor necrosis factor (TNF)-α-induced endothelial cell activation to elucidate molecular mechanism(s) of the anti-inflammatory activity of AT. AT inhibited the increase in E-selectin expression in cultured human umbilical vein endothelial cells (HUVECs) stimulated with TNF-α. In contrast, chemically modified AT that lacks affinity for heparin did not. AT inhibited the TNF-αinduced interaction of NF-κB p65 with p300, a homologue of cAMP-responsive element binding protein (CREB)-binding protein (CBP). AT increased both intracellular levels of cAMP and binding of phosphorylated-CREB to DNA in HUVECs. Forskolin showed the inhibitory effect similar to that of AT and pretreatment of HUVECs with KT-5720, an inhibitor of protein kinase A, reversed the inhibitory effect of AT. These observations suggested that AT inhibited the TNF-α-induced increase in E-selectin expression in HUVECs by inhibiting the interaction of NF-κB with CBP/p300 through cAMP-dependent protein kinase A-induced CREB activation. This inhibitory activity of AT might depend on its binding to heparin-like substances on the endothelial cell. Such an inhibitory effect of AT on TNF-α-induced endothelial cell activation might at least partly contribute to its anti-inflammatory activity.

Keywords

Antithrombin - E-selectin - cAMP - protein kinase A - CREB-binding protein/p300

Full Text

References

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