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Am J Physiol Lung Cell Mol Physiol (March 7, 2003). doi:10.1152/ajplung.00311.2002
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Submitted on September 13, 2002
Accepted on January 30, 2003

Role of Sphingosine-1 Phosphate in the Enhancement of Endothelial Barrier Integrity by Platelet-Released Products

Kane L. Schaphorst1*, Eddie Chiang1, Keri N. Jacobs1, Ari Zaiman1, Viswanathan Natarajan1, Frederick Wigley1, and Joe G. Garcia1

1 Division of Pulmonary and Critical Care Medicine, Johns Hopkins University School of Medicine, Baltimore, MD, USA

* To whom correspondence should be addressed. E-mail: kschap{at}jhmi.edu.

In vitro and in vivo evidence indicates that circulating platelets affect both vascular integrity and hemostasis. How platelets enhance the permeability barrier of the vascular endothelium is not well understood. We measured the effect of isolated human platelets on human pulmonary artery endothelial cell (EC) barrier integrity by monitoring transmonolayer electrical resistance. EC barrier function was significantly increased by the addition of platelets(~40% maximum, 2.5 x 106 platelets/ml). Platelet supernatants, derived from 2.5 x 106 platelets/ml, reproduced the barrier enhancement and reversed the barrier dysfunction produced by the edemagenic agonist thrombin, implicating a soluble barrier-promoting factor. The barrier-enhancing effect of platelet supernatants was heat stable, but attenuated by either charcoal delipidation (suggesting a vasoactive lipid mediator) or pertussis toxin, implying involvement of a Gi{alpha}-coupled receptor signal transduction pathway. Sphingosine-1-phosphate (Sph 1-P), a sphingolipid released from activated platelets, is known to ligate G-protein-coupled Edg(endothelial cell differentiation gene) receptors, increase EC electrical resistance, and reorganize the actin cytoskeleton (J Clin Invest 2001; 108:689-701). Infection of EC with an adenoviral vector expressing an antisense oligonucleotide directed against Edg-1, but not infection with control vector, attenuated the barrier-enhancing effect of both platelet supernatants and Sph 1-P. These results indicate that a major physiologically relevant vascular barrier-protective mediator produced by human platelets is sphingosine-1-phosphate.




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