Title
Transient and prolonged increase in endothelial permeability induced by histamine and thrombin: Role of protein kinases, calcium, and RhoA
Author
van Nieuw Amerongen, G.P.
Draijer, R.
Vermeer, M.A.
van Hinsbergh, V.W.M.
Gaubius Instituut TNO
Publication year
1998
Abstract
In the present study, we differentiated between short- and long-term effects of vasoactive compounds on human endothelial permeability in an in vitro model. Histamine induced a rapid and transient (<3 minutes) decrease in barrier function, as evidenced by a decreased transendothelial electrical resistance and an increased passage of 22Na ions. This increase in permeability was inhibited completely by chelation of intracellular calcium ions by BAPTA-AM and inhibition of calmodulin activity and myosin light chain (MLC) phosphorylation. The presence of serum factors prolonged the barrier dysfunction induced by histamine. Thrombin by itself induced a prolonged barrier dysfunction (>30 minutes) as evidenced by an increased passage of peroxidase and 40 kDa dextran. It was dependent only partially on calcium ions and calmodulin. The protein tyrosine kinase inhibitors genistein and herbimycin A, but not the inactive analogue daidzein, inhibited to a large extent the increase in permeability induced by thrombin. Genistein and BAPTA- AM inhibited the thrombin-induced permeability in an additive way, causing together an almost complete prevention of the thrombin-induced increase in permeability. Inhibition of protein tyrosine kinase was accompanied by a decrease in MLC phosphorylation and a reduction in the extent of F-actin fiber and focal attachment formation. Inhibition of RhoA by C3 transferase toxin reduced both the thrombin-induced barrier dysfunction and MLC phosphorylation. Genistein and C3 transferase toxin did not elevate the cellular cAMP levels. No evidence was found for a significant role of protein kinase C in the thrombin-induced increase in permeability or in the accompanying MLC phosphorylation. These data indicate that in endothelial cell monolayers that respond to histamine in a physiological way, thrombin induces a prolonged increase in permeability by 'calcium sensitization,' which involves protein tyrosine phosphorylation and RhoA activation.
Subject
Biology
Human endothelial cell
Protein kinase C
Protein tyrosine kinase
RhoA
Calcium
Capillary Permeability
Cells, Cultured
Cytoskeleton
Endothelium, Vascular
GTP-Binding Proteins
Histamine
Humans
Infant, Newborn
Protein Kinases
Protein-Tyrosine Kinases
rhoA GTP-Binding Protein
Thrombin
Time Factors
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DOI
https://doi.org/10.1161/01.res.83.11.1115
TNO identifier
234688
ISSN
0009-7330
Source
Circulation Research, 83 (11), 1115-1123
Document type
article