Arteriosclerosis, Thrombosis, and Vascular Biology. 1995;15:2055-2062
© 1995 American Heart Association, Inc.
Articles |
Thrombin Induces the Redistribution and Acute Release of Tissue Factor Pathway Inhibitor From Specific Granules Within Human Endothelial Cells in Culture
From the Thrombosis Research Institute, Chelsea, London.
Correspondence to Dr Cristina Lupu, Coagulation and Fibrinolysis Section, Thrombosis Research Institute, Emmanuel Kaye Building, Manresa Rd, Chelsea, London SW3 6LR UK.
Abstract Tissue factor pathway inhibitor (TFPI) is a vascular anticoagulant that regulates the tissue factor (TF)–dependent pathway of coagulation. The majority of intravascular TFPI is thought to be noncovalently bound to the vessel wall. Our immunolocalization studies in cultures of human umbilical vein endothelial cells (HUVEC) and immortalized EA.hy926 cells show that TFPI is located in well-defined granules evenly spread over the cell surface and with apical polarization within the cytoplasm. These granules are smaller than and distinct from Weibel-Palade bodies. Upon treatment of cultured cells with low concentrations of thrombin (0.01 to 1 NIH U/mL), a marked redistribution of TFPI, occurred with patching in focal points and increased exposure of both TFPI antigen and anticoagulant activity on the surface of the stimulated endothelial cells. This redistribution was paralleled by an acute release of TFPI in the cell medium. EA.hy926 cells responded more readily to thrombin stimulation than HUVECs. The process was inhibited by both hirudin and anti-thrombin receptor antibody. Our findings demonstrate a novel mechanism by which thrombin may exert a negative feedback control on blood coagulation. Therefore, this pathway can be of physiological importance in controlling TF-mediated thrombin generation.
Key Words: tissue factor pathway inhibitor • endothelial cells • thrombin • confocal microscopy
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