Premium
Effect of pigment epithelial derived factor (PEDF) on vascular endothelial growth factor (VEGF)‐mediated endothelial cell (EC) permeability
Author(s) -
Shin Meyeon,
Duh Elia J,
Sham James,
He Chaoxia,
Gurkan Ozlem U,
Becker Patrice M
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.6.a1432
Rationale: VEGF is a potent mediator of enhanced vascular permeability. PEDF, a 50 kDa non‐inhibitory serpin, was shown to inhibit both VEGF‐induced retinal angiogenesis and vascular hyperpermeability in vivo . Methods: To investigate mechanisms by which PEDF may inhibit VEGF‐induced EC permeability, we established an in vitro model utilizing porcine aortic EC stably expressing both VEGF receptor 2 (VEGFR2) and neuropilin‐1. The effects of PEDF (10, 50 and 100 mM) on VEGF‐mediated EC barrier dysfunction were determined by measurement of transendothelial electrical resistance (TER) using electrical cell‐substrate impedance sensing (n=5–7/group). Effects of PEDF on VEGF‐induced EC VEGFR2 activation were determined by Western blot and confocal immunohistochemistry (IHC) for phospho‐VEGFR2 expression (n=3). Results: The effects of PEDF on decreased TER in response to VEGF (100ng/ml) were dose‐dependent. Pretreatment with either 50 or 100 mM PEDF significantly attenuated VEGF‐induced decreased TER for ≥ 3 hr (p<0.005), while 10 mM PEDF had no effect on this response. Preliminary Western blot and IHC analysis suggest PEDF (100mM) may partially attenuate VEGFR2 phosphorylation after administration of VEGF. Conclusions: These data support a role for PEDF in the inhibition of VEGF‐mediated enhanced EC permeability, and suggest that PEDF may act by altering VEGFR2 signaling. Funded by NIH HL083286