High affinity binding sites for 12(R)-Hydroxyeicosatrienoic acid [12(R)-HETrE] in microvessel endothelial cells.
J Ocul Pharmacol Ther
; 13(3): 191-9, 1997 Jun.
Article
em En
| MEDLINE
| ID: mdl-9185033
ABSTRACT
12(R)-HETrE is an NADPH-dependent arachidonic acid-derived metabolite whose synthesis is induced several fold in inflamed corneal epithelium correlating with the development of the in situ inflammatory response, i.e., vasodilation, PMN chemotaxis, endothelial cell mitogenesis, and neovascularization. Because this novel eicosanoid may serve as an endogenous mediator of the angiogenic response in the cornea during inflammation we probed microvessel endothelial cells for a specific binding site which could possibly account for the mechanism by which this eicosanoid initiates changes in cellular activity. Binding of radioactive ligand [3H-12(R)-HETrE] was saturable with time and concentration. Scatchard analysis indicated a single, saturable binding site for 12(R)-HETrE with a Bmax = 24,700 sites/cell and an apparent Kd = 0.043 nM. Thin layer chromatography analysis of cell-associated ligand revealed that esterification of 12(R)-HETrE was 2-7 fold less than unesterified, cell bound ligand. The concentrations of 12(R)-HETrE at which maximum biological activity has been observed, i.e., 0.1 nM, roughly corresponds to the Kd value, suggesting a functional link to this binding site. These studies begin to reveal a potential mechanism by which 12(R)-HETrE stimulates microvessel endothelial cells to invade the cornea leading to corneal neovascularization.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Sítios de Ligação
/
Endotélio Vascular
/
Ácido 12-Hidroxi-5,8,10,14-Eicosatetraenoico
Limite:
Animals
Idioma:
En
Revista:
J Ocul Pharmacol Ther
Assunto da revista:
FARMACOLOGIA
/
OFTALMOLOGIA
/
TERAPEUTICA
Ano de publicação:
1997
Tipo de documento:
Article
País de afiliação:
Estados Unidos