Prostaglandin D2-DP signaling promotes endothelial barrier function via the cAMP/PKA/Tiam1/Rac1 pathway

Arterioscler Thromb Vasc Biol. 2013 Mar;33(3):565-71. doi: 10.1161/ATVBAHA.112.300993. Epub 2013 Jan 10.

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

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adherens Junctions / drug effects
  • Adherens Junctions / enzymology
  • Animals
  • Capillary Permeability* / drug effects
  • Cells, Cultured
  • Cyclic AMP / metabolism*
  • Cyclic AMP-Dependent Protein Kinases / metabolism*
  • Cytoskeleton / drug effects
  • Cytoskeleton / enzymology
  • Dextrans / metabolism
  • Dose-Response Relationship, Drug
  • Ear Auricle / blood supply*
  • Electric Impedance
  • Enzyme Activation
  • Fluorescein-5-isothiocyanate / analogs & derivatives
  • Fluorescein-5-isothiocyanate / metabolism
  • Guanine Nucleotide Exchange Factors / genetics
  • Guanine Nucleotide Exchange Factors / metabolism*
  • Human Umbilical Vein Endothelial Cells / drug effects
  • Human Umbilical Vein Endothelial Cells / enzymology*
  • Humans
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Prostaglandin D2 / metabolism*
  • Protein Kinase Inhibitors / pharmacology
  • RNA Interference
  • Receptors, Immunologic / agonists
  • Receptors, Immunologic / deficiency
  • Receptors, Immunologic / genetics
  • Receptors, Immunologic / metabolism*
  • Receptors, Prostaglandin / agonists
  • Receptors, Prostaglandin / deficiency
  • Receptors, Prostaglandin / genetics
  • Receptors, Prostaglandin / metabolism*
  • Signal Transduction* / drug effects
  • T-Lymphoma Invasion and Metastasis-inducing Protein 1
  • Time Factors
  • Transfection
  • cdc42 GTP-Binding Protein / metabolism
  • rac1 GTP-Binding Protein / metabolism*

Substances

  • Dextrans
  • Guanine Nucleotide Exchange Factors
  • Protein Kinase Inhibitors
  • RAC1 protein, human
  • RAPGEF3 protein, human
  • Receptors, Immunologic
  • Receptors, Prostaglandin
  • T-Lymphoma Invasion and Metastasis-inducing Protein 1
  • TIAM1 protein, human
  • fluorescein isothiocyanate dextran
  • Cyclic AMP
  • Cyclic AMP-Dependent Protein Kinases
  • cdc42 GTP-Binding Protein
  • rac1 GTP-Binding Protein
  • Fluorescein-5-isothiocyanate
  • Prostaglandin D2
  • prostaglandin D2 receptor