Abstract
Objective:
Prostaglandin D(2) (PGD(2)) is one of the prostanoids produced during inflammation. Although PGD(2) is known to decrease endothelial permeability through D prostanoid (DP) receptor stimulation, the detailed mechanism is unknown.
Methods and results:
Treatment with PGD(2) (0.1-3 μmol/L) or the DP receptor agonist, BW245C (0.1-3 μmol/L), dose-dependently increased transendothelial electrical resistance and decreased the FITC-dextran permeability of human umbilical vein endothelial cells. Both indicated decreased endothelial permeability. These phenomena were accompanied by Tiam1/Rac1-dependent cytoskeletal rearrangement. BW245C (0.3 μmol/L) increased the intracellular cAMP level and subsequent protein kinase A (PKA) activity. Pretreatment with PKA inhibitory peptide, but not gene depletion of exchange protein directly activated by cAMP 1 (Epac1), attenuated BW245C-induced Rac1 activation and transendothelial electric resistance increase. In vivo, application of 2.5% croton oil or histamine (100 μg) caused vascular leakage indexed by dye extravasation. Pretreatment with BW245C (1 mg/kg) attenuated the dye extravasation. Gene deficiency of DP abolished, or inhibition of PKA significantly reduced, the DP-mediated barrier enhancement.
Conclusions:
PGD(2)-DP signaling reduces vascular permeability both in vivo and in vitro. This phenomenon is mediated by cAMP/PKA/Tiam1-dependent Epac1-independent Rac1 activation and subsequent enhancement of adherens junction in endothelial cell.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adherens Junctions / drug effects
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Adherens Junctions / enzymology
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Animals
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Capillary Permeability* / drug effects
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Cells, Cultured
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Cyclic AMP / metabolism*
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Cyclic AMP-Dependent Protein Kinases / metabolism*
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Cytoskeleton / drug effects
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Cytoskeleton / enzymology
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Dextrans / metabolism
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Dose-Response Relationship, Drug
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Ear Auricle / blood supply*
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Electric Impedance
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Enzyme Activation
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Fluorescein-5-isothiocyanate / analogs & derivatives
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Fluorescein-5-isothiocyanate / metabolism
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Guanine Nucleotide Exchange Factors / genetics
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Guanine Nucleotide Exchange Factors / metabolism*
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Human Umbilical Vein Endothelial Cells / drug effects
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Human Umbilical Vein Endothelial Cells / enzymology*
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Humans
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Mice
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Mice, Inbred C57BL
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Mice, Knockout
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Prostaglandin D2 / metabolism*
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Protein Kinase Inhibitors / pharmacology
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RNA Interference
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Receptors, Immunologic / agonists
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Receptors, Immunologic / deficiency
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Receptors, Immunologic / genetics
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Receptors, Immunologic / metabolism*
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Receptors, Prostaglandin / agonists
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Receptors, Prostaglandin / deficiency
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Receptors, Prostaglandin / genetics
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Receptors, Prostaglandin / metabolism*
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Signal Transduction* / drug effects
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T-Lymphoma Invasion and Metastasis-inducing Protein 1
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Time Factors
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Transfection
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cdc42 GTP-Binding Protein / metabolism
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rac1 GTP-Binding Protein / metabolism*
Substances
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Dextrans
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Guanine Nucleotide Exchange Factors
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Protein Kinase Inhibitors
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RAC1 protein, human
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RAPGEF3 protein, human
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Receptors, Immunologic
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Receptors, Prostaglandin
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T-Lymphoma Invasion and Metastasis-inducing Protein 1
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TIAM1 protein, human
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fluorescein isothiocyanate dextran
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Cyclic AMP
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Cyclic AMP-Dependent Protein Kinases
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cdc42 GTP-Binding Protein
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rac1 GTP-Binding Protein
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Fluorescein-5-isothiocyanate
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Prostaglandin D2
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prostaglandin D2 receptor