Antipermeability function of PEDF involves blockade of the MAP kinase/GSK/β-catenin signaling pathway and uPAR expression

Jinling Yang, Elia J. Duh, Ruth B Caldwell, M. Ali Behzadian

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Purpose. Pigment epithelium- derived factor (PEDF) is a potent inhibitor of vascular endothelial growth factor (VEGF)-induced endothelial permeability. The goal of this study was to understand the mechanism by which PEDF blocks VEGF-induced increases in vascular permeability. Methods. The paracellular permeability of bovine retinal endothelial (BRE) cells was measured by assaying transendothelial cell electrical resistance and tracer flux. Western blot analysis was used to show phosphorylation of VEGFR2, MAP kinases, and glycogen synthase kinase 3 (GSK3)-β. Confocal imaging and Western blot analysis were used to determine subcellular distribution of β-catenin. Real-time RT-PCR and Western blot analysis were used to quantify urokinase plasminogen activator receptor (uPAR) expression. Results. PEDF blocked VEGF-induced phosphorylation of extracellular signal-regulated kinase (ERK), p38 MAP kinase, the p38 substrate MAP kinase-activated protein kinase-2 (MAP-KAPK-2), and GSK3-β, but it had no effect on the phosphorylation of VEGFR2. In addition, the VEGF-induced transcriptional activation of β-catenin and uPAR expression were blocked by PEDF and by inhibitors of p38 and MEK. Finally, the VEGF-induced increase in permeability was blocked by both PEDF and the same kinase inhibitors. Conclusions. The data suggest that p38 MAP kinase and ERK act upstream of GSK/β-catenin in VEGF-induced activation of the uPA/uPAR system and that PEDF-mediated inhibition of the VEGF-induced increase in vascular permeability involves blockade of this pathway. These findings are important for developing precise and potent therapies for treatment of diseases characterized by vascular barrier dysfunction.

Original languageEnglish (US)
Pages (from-to)3273-3280
Number of pages8
JournalInvestigative Ophthalmology and Visual Science
Volume51
Issue number6
DOIs
StatePublished - Jun 1 2010

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Urokinase Plasminogen Activator Receptors
Catenins
Vascular Endothelial Growth Factor A
Phosphotransferases
p38 Mitogen-Activated Protein Kinases
Glycogen Synthase Kinase 3
Permeability
Western Blotting
Mitogen-Activated Protein Kinase Kinases
Extracellular Signal-Regulated MAP Kinases
Phosphorylation
Capillary Permeability
pigment epithelium-derived factor
Electric Impedance
Vascular Diseases
Transcriptional Activation
Real-Time Polymerase Chain Reaction
Endothelial Cells

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

Antipermeability function of PEDF involves blockade of the MAP kinase/GSK/β-catenin signaling pathway and uPAR expression. / Yang, Jinling; Duh, Elia J.; Caldwell, Ruth B; Ali Behzadian, M.

In: Investigative Ophthalmology and Visual Science, Vol. 51, No. 6, 01.06.2010, p. 3273-3280.

Research output: Contribution to journalArticle

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abstract = "Purpose. Pigment epithelium- derived factor (PEDF) is a potent inhibitor of vascular endothelial growth factor (VEGF)-induced endothelial permeability. The goal of this study was to understand the mechanism by which PEDF blocks VEGF-induced increases in vascular permeability. Methods. The paracellular permeability of bovine retinal endothelial (BRE) cells was measured by assaying transendothelial cell electrical resistance and tracer flux. Western blot analysis was used to show phosphorylation of VEGFR2, MAP kinases, and glycogen synthase kinase 3 (GSK3)-β. Confocal imaging and Western blot analysis were used to determine subcellular distribution of β-catenin. Real-time RT-PCR and Western blot analysis were used to quantify urokinase plasminogen activator receptor (uPAR) expression. Results. PEDF blocked VEGF-induced phosphorylation of extracellular signal-regulated kinase (ERK), p38 MAP kinase, the p38 substrate MAP kinase-activated protein kinase-2 (MAP-KAPK-2), and GSK3-β, but it had no effect on the phosphorylation of VEGFR2. In addition, the VEGF-induced transcriptional activation of β-catenin and uPAR expression were blocked by PEDF and by inhibitors of p38 and MEK. Finally, the VEGF-induced increase in permeability was blocked by both PEDF and the same kinase inhibitors. Conclusions. The data suggest that p38 MAP kinase and ERK act upstream of GSK/β-catenin in VEGF-induced activation of the uPA/uPAR system and that PEDF-mediated inhibition of the VEGF-induced increase in vascular permeability involves blockade of this pathway. These findings are important for developing precise and potent therapies for treatment of diseases characterized by vascular barrier dysfunction.",
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AU - Ali Behzadian, M.

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N2 - Purpose. Pigment epithelium- derived factor (PEDF) is a potent inhibitor of vascular endothelial growth factor (VEGF)-induced endothelial permeability. The goal of this study was to understand the mechanism by which PEDF blocks VEGF-induced increases in vascular permeability. Methods. The paracellular permeability of bovine retinal endothelial (BRE) cells was measured by assaying transendothelial cell electrical resistance and tracer flux. Western blot analysis was used to show phosphorylation of VEGFR2, MAP kinases, and glycogen synthase kinase 3 (GSK3)-β. Confocal imaging and Western blot analysis were used to determine subcellular distribution of β-catenin. Real-time RT-PCR and Western blot analysis were used to quantify urokinase plasminogen activator receptor (uPAR) expression. Results. PEDF blocked VEGF-induced phosphorylation of extracellular signal-regulated kinase (ERK), p38 MAP kinase, the p38 substrate MAP kinase-activated protein kinase-2 (MAP-KAPK-2), and GSK3-β, but it had no effect on the phosphorylation of VEGFR2. In addition, the VEGF-induced transcriptional activation of β-catenin and uPAR expression were blocked by PEDF and by inhibitors of p38 and MEK. Finally, the VEGF-induced increase in permeability was blocked by both PEDF and the same kinase inhibitors. Conclusions. The data suggest that p38 MAP kinase and ERK act upstream of GSK/β-catenin in VEGF-induced activation of the uPA/uPAR system and that PEDF-mediated inhibition of the VEGF-induced increase in vascular permeability involves blockade of this pathway. These findings are important for developing precise and potent therapies for treatment of diseases characterized by vascular barrier dysfunction.

AB - Purpose. Pigment epithelium- derived factor (PEDF) is a potent inhibitor of vascular endothelial growth factor (VEGF)-induced endothelial permeability. The goal of this study was to understand the mechanism by which PEDF blocks VEGF-induced increases in vascular permeability. Methods. The paracellular permeability of bovine retinal endothelial (BRE) cells was measured by assaying transendothelial cell electrical resistance and tracer flux. Western blot analysis was used to show phosphorylation of VEGFR2, MAP kinases, and glycogen synthase kinase 3 (GSK3)-β. Confocal imaging and Western blot analysis were used to determine subcellular distribution of β-catenin. Real-time RT-PCR and Western blot analysis were used to quantify urokinase plasminogen activator receptor (uPAR) expression. Results. PEDF blocked VEGF-induced phosphorylation of extracellular signal-regulated kinase (ERK), p38 MAP kinase, the p38 substrate MAP kinase-activated protein kinase-2 (MAP-KAPK-2), and GSK3-β, but it had no effect on the phosphorylation of VEGFR2. In addition, the VEGF-induced transcriptional activation of β-catenin and uPAR expression were blocked by PEDF and by inhibitors of p38 and MEK. Finally, the VEGF-induced increase in permeability was blocked by both PEDF and the same kinase inhibitors. Conclusions. The data suggest that p38 MAP kinase and ERK act upstream of GSK/β-catenin in VEGF-induced activation of the uPA/uPAR system and that PEDF-mediated inhibition of the VEGF-induced increase in vascular permeability involves blockade of this pathway. These findings are important for developing precise and potent therapies for treatment of diseases characterized by vascular barrier dysfunction.

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