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Apoptotic vascular endothelial cells become procoagulant
T Bombeli, A Karsan, JF Tait and JM Harlan
Department of Laboratory Medicine, University of Washington, Seattle 98195,
USA.
Whereas unperturbed endothelial cells provide potent anticoagulant
properties, exposure to inflammatory and atherogenic stimuli can rapidly
lead to a procoagulant behavior. Because recent studies provide evidence
that apoptosis of vascular cells may occur under conditions such as
atherosclerosis and inflammation, we investigated whether apoptotic
endothelial cells may contribute to the development of a prothrombotic
state. In this report, it is shown that both adherent and detached
apoptotic human umbilical vein endothelial cells (HUVECs) become
procoagulant. Apoptosis was induced by staurosporine, a nonspecific protein
kinase inhibitor, or by culture in suspension with serum deprivation. Both
methods resulted in similar findings. As assessed by flow cytometric
determination of annexin V binding, HUVECs undergoing cell death exhibited
typically a more rapid exposure of membrane phosphatidylserine (PS) than
DNA fragmentation. Depending on the stage of apoptosis, this redistribution
of phospholipids was found to induce an increase of the activity of the
intrinsic tenase complex by 25% to 60%. Although apoptotic cells did not
show antigenic or functional tissue factor (TF) activity, when preactivated
with lipopolysaccharide, TF procoagulant activity increased by 50% to 70%.
At 8 hours after apoptosis induction, antigenic thrombomodulin, heparan
sulfates, and TF pathway inhibitor decreased by about 83%, 80%, and 59%,
respectively. The functional activity of these components was reduced by
about 36%, 52%, and 39%, respectively. Moreover, the presence of apoptotic
HUVECs led to a significant increase of thrombin formation in recalcified
citrated plasma. In conclusion, apoptotic HUVECs, either adherent or in
suspension, become procoagulant by increased expression of PS and the loss
of anticoagulant membrane components.
Volume 89,
Issue 7,
pp. 2429-2442,
04/01/1997
Copyright © 1997 by The American Society of Hematology
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