Fibroblast Growth Factor-2 Mediates Angiotensin-Converting Enzyme Inhibitor-Induced Angiogenesis in Coronary Endothelium

  1. Sandra Donnini,
  2. Raffaella Solito,
  3. Antonio Giachetti,
  4. Harris J. Granger,
  5. Marina Ziche and
  6. Lucia Morbidelli
  1. Section of Pharmacology, Department of Molecular Biology, University of Siena, Siena, Italy (S.D., R.S., A.G., M.Z., L.M.); and Cardiovascular Research Institute, Department of Medical Physiology, College of Medicine, Texas A&M University System Health Science Center, Temple, Texas (H.J.G.)
  1. Address correspondence to:
    Dr. Lucia Morbidelli, Pharmacology Section, Department of Molecular Biology, Via A. Moro 2, 53100 Siena, Italy. E-mail: morbidelli{at}unisi.it

Abstract

The beneficial effect exerted by angiotensin-converting enzyme inhibitors (ACEI) on vascular endothelium has been attributed to restoration of endothelial cell survival properties and improvement of angiogenesis. Fibroblast growth factor (FGF)-2 is an angiogenic factor for the microvascular endothelium, which tonically promotes endothelial cell growth and survival through an autocrine/paracrine mechanism. Here, we formulate the hypothesis that FGF-2 might contribute to the prosurvival/proangiogenic effect of ACEI. We investigated zofenoprilat and, in selected experiments, lisinopril, as representatives of ACEI. These compounds induced formation of pseudocapillaries in vessel fragments isolated from porcine coronary and human umbilical arteries by increasing endothelial cell growth up to 5-fold. Angiogenesis was abolished by inhibitors of nitric-oxide synthase (NOS) pathway and by anti-FGF-2 antibodies. Consistently, in cultured coronary endothelial cells (CVECs), ACEI up-regulated endothelial NOS (eNOS) and FGF-2 and induced mitogen-activated protein kinase extracellular signal-regulated kinase 1/2 activation. The overexpression of eNOS/FGF-2 produced, at the functional level, enhanced cell proliferation and migration, the latter effect being dose-dependent and maximal at 0.1 μM zofenoprilat. The importance of FGF-2 for the acquisition of the angiogenic phenotype elicited by ACEI was clearly demonstrated by the impairment of endothelial functions following transfection of CVECs with small interference RNA for FGF-2. Moreover, FGF-2 silencing greatly affected the nuclear translocation of the FGF receptor (FGFR)-1, highlighting the autocrine mode of action of FGF-2. At the endothelial membrane level, zofenoprilat appeared to activate the bradykinin B1 receptor, a known stimulant of FGF-2 expression. In conclusion, we show that ACEI exert protective/proangiogenic effects in microvascular coronary endothelial cells by activating the endogenous FGF-2/FGFR-1 system.

Footnotes

  • This work was supported by the Italian Ministry for Research (Ministero dell'Istruzione, dell'Università e della Ricerca, Programmi di Ricerca di Interesse Nazionale Project 2004065317_001 and Fondo per gli Investimenti della Ricerca di Base Project RBNE01A882_002), by the Progetto Regione Toscana “Advanced Methods for the Growth and Differentiation of Adult Staminal Cells to Develop Cell Therapies,” and by European Community Sixth Frame-work Programme funding (LSHM-CT-2004-0050333).

  • Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.

  • doi:10.1124/jpet.106.108803.

  • ABBREVIATIONS: ACEI, angiotensin-converting enzyme inhibitor(s); NOS, nitric-oxide synthase; eNOS, endothelial nitric-oxide synthase; FGF, fibroblast growth factor; FGFR, FGF receptor; ACE, angiotensin-converting enzyme; MAPK, mitogen-activated protein kinase; ERK, extracellular signal-regulated kinase; PCA, porcine coronary artery; HUA, human umbilical artery; CVEC, coronary venular endothelial cell; BK, bradykinin; CS, calf serum; MEK, mitogen-activated protein kinase kinase; U0126, 1,4-diamino-2,3-dicyano-1,4-bis[2-aminophenylthio]butadiene; PBS, phosphate-buffered saline; BSA, bovine serum albumin; RT, reverse transcriptase; PCR, polymerase chain reaction; FCS, fetal calf serum; l-NMMA, Nω-methyl-l-arginine; ODQ, 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one; SU5402, 3-[3-(2-carboxyethyl)-4-methylpyrrol-2-methylidenyl]-2-indolinone; siRNA, small interference RNA.

    • Received June 7, 2006.
    • Accepted July 21, 2006.
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  1. Published online before print July 25, 2006, doi: 10.1124/jpet.106.108803 JPET November 2006 vol. 319 no. 2 515-522
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