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1 Department of Anesthesiology, Aichi Medical University School of Medicine, Aichi-gun, Aichi, Japan
2 Department of Anesthesiology, Nagoya University School of Medicine, Nagoya, Aichi, Japan
3 Department of Pharmacology, Aichi Medical University School of Medicine, Aichi-gun, Aichi, Japan
4 Department of Anatomy, Aichi Medical University School of Medicine, Aichi-gun, Aichi, Japan
* To whom correspondence should be addressed. E-mail: nao{at}aichi-med-u.ac.jp.
Neuropeptide Y (NPY) elevates permeability of cultured rat aortic endothelial cells (RAECs) in monolayer culture under hypoxic conditions (5 % O2), possibly through binding to the NPY Y3 receptor. The present study was undertaken to evaluate the effects of NPY in comparison with vascular endothelial growth factor (VEGF). RAECs were cultured on the upper chamber base in a double chamber culture system, into which FITC-labeled albumin was introduced, and permeation into lower chamber was measured. Treatment was with 3 x 10 -7 M NPY or 10-7 g/mL VEGF, for 2 h, along with specific inhibitors. VEGFR-2 tyrosine kinase inhibitor Tyrphostin SU1498 and protein kinase C inhibitor GF-109203X suppressed the VEGF-induced increase in monolayer permeability, but not that caused by NPY. Furthermore, while the action of NPY was concentration-dependently blocked by phospholipase C inhibitor U-73122, it was less sensitive than VEGF. However, effects of both NPY and VEGF on RAEC monolayer permeability were blocked with equal concentration-dependence by STI571, an inhibitor of Abl tyrosine kinase in nucleus and/or cytoplasm. The myosin light chain kinase inhibitor ML-9 suppressed both NPY- and VEGF-induced increment in permeability by approximately 70 %, while the calmodulin-dependent kinase inhibitor DY9760e could decrease to below the base line. These results indicate that the NPY Y3-receptor subtype is specifically linked to the effects of STI571 on endothelial cells, and that NPY, a sympathetic co-neurotransmitter, may increase vascular permeability, in association with altered intracellular or nuclear signal transduction.
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