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Thrombin-induced gap formation in confluent endothelial cell monolayers in
vitro
M Laposata, DK Dovnarsky and HS Shin
When thrombin is incubated with confluent monolayers of human umbilical
vein endothelial cells in vitro, there is a change in the shape of the
endothelial cells that results in gaps in the monolayer, disrupting the
integrity of the endothelium and exposing the subendothelium. Using a grid
assay to measure this phenomenon, we observed that up to 80% of the surface
area once covered by cells was uncovered after a 15-min incubation with
10(-2) U/ml (10(-10)M) thrombin. The effect was apparent within 2 min and
did not remove cells from the surface of the culture dish. The gaps in the
monolayer completely disappeared within 2 hr after exposure to thrombin.
The effect of thrombin was inhibited by preincubation of thrombin with
hirudin or antithrombin III plus heparin or by preincubation of the
monolayers with dibutyryl cyclic adenosine monophosphate (dbcAMP).
Histamine also induced gap formation in endothelial cell monolayers. Both
pyrilamine and cimetidine prevented the histamine-induced effect, but they
had no effect on thrombin- induced gap formation. Intact monolayers were
not disrupted by bradykinin, serotonin, C5a, or C3a. Our results suggest
that small amounts of thrombin can induce repeated and transient exposure
of the subendothelium, a situation believed to be conducive to
atherogenesis and thrombosis.
Volume 62,
Issue 3,
pp. 549-556,
09/01/1983
Copyright © 1983 by The American Society of Hematology
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