Endothelium-derived nitric oxide (NO) activates the NO-epidermal growth factor receptor-mediated signaling pathway in bradykinin-stimulated angiogenesis

Autor(es)

Moraes, Miriam S. ^[1] ; Costa, Paulo E. ^[2] ; Batista, Wagner L. ^{[3, 4]} ; Paschoalin, Taysa ^[4] ; Curcio, Marli F. ^{[2, 4]} ; Borges, Roberta E. ^[2] ; Taha, Murched O. ^[5] ; Fonseca, Fabio V. ^[6] ; Stern, Arnold ^{[7, 8]} ; Monteiro, Hugo P. ^[2]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Inst Quim, Dept Biochem, Sao Paulo - Brazil

^[2] Univ Fed Sao Paulo, Ctr Cellular & Mol Therapy CTCMol, Dept Biochem, BR-04044010 Sao Paulo - Brazil

^[3] Univ Fed Sao Paulo, Dept Biol Sci, Sao Paulo - Brazil

^[4] Univ Fed Sao Paulo, Dept Microbiol Immunol & Parasitol, BR-04044010 Sao Paulo - Brazil

^[5] Univ Fed Sao Paulo, Dept Surg, BR-04044010 Sao Paulo - Brazil

^[6] Case Western Univ, Inst Transformat Mol Med, Dept Med, Cleveland, OH - USA

^[7] NYU, Sch Med, Dept Pharmacol, New York, NY - USA

^[8] Univ Spiritu Santo Ecuador, Sch Med, Guayaquil - Ecuador

• Tipo de documento: Artigo Científico
• Fonte: Archives of Biochemistry and Biophysics; v. 558, p. 14-27, SEP 15 2014.
• Citações Web of Science: 27

Resumo

Nitric oxide (NO) is involved in angiogenesis and stimulates the EGF-R signaling pathway. Stimulation of different endothelial cell lines with bradykinin (BK) activates the endothelial NO synthase (eNOS) and promotes EGF-R tyrosine phosphorylation. Increase in NO production correlated with enhanced phosphorylation of tyrosine residues and S-nitrosylation of the EGF-R. NO-mediated stimulatory effects on tyrosine phosphorylation of the EGF-R, where cGMP independent. Inhibition of soluble guanylyl cyclase followed by BK stimulation of human umbilical vein endothelial cells (HUVECs) did not change tyrosine phosphorylation levels of EGF-R. BK-stimulation of HUVEC promoted S-nitrosylation of the phosphatase SHP-1 and of p21Ras. Phosphorylation and activation of the ERK1/2 MAP kinases mediated by BK was dependent on the activation of the B2 receptor, of the EGF-R, and of p21 Ras. Inhibition of BK-stimulated S-nitrosylation prevented the activation of the ERK1/2 MAP kinases. Furthermore, activated ERK1/2 MAP kinases inhibited internalization of EGF-R by phosphorylating specific Thr residues of its cytoplasmic domain. BK-induced proliferation of endothelial cells was partially inhibited by the NOS inhibitor (L-NAME) and by the MEK inhibitor (PD98059). BK stimulated the expression of vascular endothelial growth factor (VEGF). VEGF expression was dependent on the activation of the EGF-R, the B2 receptor, p21Ras, and on NO generation. A Matrigel (R)-based in vitro assay for angiogenesis showed that BK induced the formation of capillary-like structures in HUVEC, but not in those cells expressing a mutant of the EGF-R lacking tyrosine kinase activity. Additionally, pre-treatment of BK-stimulated HUVEC with L-NAME, PD98059, and with SU5416, a specific inhibitor of VEGFR resulted in inhibition of in vitro angiogenesis. Our findings indicate that BK-mediated angiogenesis in endothelial cells involves the induction of the expression of VEGF associated with the activation of the NO/EGF-R/p21Ras/ERK1/2 MAP kinases signaling pathway. (C) 2014 Elsevier Inc. All rights reserved. (AU)

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