title: Antagonism of bradykinin B2 receptor prevents inflammatory responses in human endothelial cells by quenching the NF-kB pathway activation.
abstract: BACKGROUND
  Bradykinin (BK) induces angiogenesis by promoting vessel permeability, growth and remodeling. This study aimed to demonstrate that the B2R antagonist, fasitibant, inhibits the BK pro-angiogenic effects.
  
  
  METHODS
  We assesed the ability of fasibitant to antagonize the BK stimulation of cultured human cells (HUVEC) and circulating pro-angiogenic cells (PACs), in producing cell permeability (paracellular flux), migration and pseocapillary formation. The latter parameter was studied in vitro (matrigel assay) and in vivo in mice (matrigel plug) and in rat model of experimental osteoarthritis (OA). We also evaluated NF-κB activation in cultured cells by measuring its nuclear translocation and its downstream effectors such as the proangiogenic ciclooxygenase-2 (COX-2), prostaglandin E-2 and vascular endothelial growth factor (VEGF).
  
  
  RESULTS
  HUVEC, exposed to BK (1-10 µM), showed increased permeability, disassembly of adherens and tight-junction, increased cell migration, and pseudocapillaries formation. We observed a significant increase of vessel density in the matrigel assay in mice and in rats OA model. Importantly, B2R stimulation elicited, both in HUVEC and PACs, NF-κB activation, leading to COX-2 overexpression, enhanced prostaglandin E-2 production. and VEGF output. The BK/NF-κB axis, and the ensuing amplification of inflammatory/angiogenic responses were fully prevented by fasitibant as well as by IKK VII, an NF-κB. Inhibitor.
  
  
  CONCLUSIONS
  This work illustrates the role of the endothelium in the inflammation provoked by the BK/NF-κB axis. It also demonstates that B2R blockade by the antaogonist fasibitant, abolishes both the initial stimulus and its amplification, strongly attenuating the propagation of inflammation.
authors.affiliation: Department of Life Sciences, University of Siena, Siena, Italy¸
authors.forename: Erika
authors.initials: E
authors.lastname: Terzuoli
authors.affiliation: Pharmacology Department, Menarini Ricerche S.p.A, Florence, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Meini
authors.affiliation: Pharmacology Department, Menarini Ricerche S.p.A, Florence, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Cucchi
authors.affiliation: Pharmacology Department, Menarini Ricerche S.p.A, Florence, Italy.
authors.forename: Claudio
authors.initials: C
authors.lastname: Catalani
authors.affiliation: Pharmacology Department, Menarini Ricerche S.p.A, Florence, Italy.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Cialdai
authors.affiliation: Pharmacology Department, Menarini Ricerche S.p.A, Florence, Italy.
authors.forename: Carlo Alberto
authors.initials: CA
authors.lastname: Maggi
authors.affiliation: Department of Life Sciences, University of Siena, Siena, Italy¸
authors.forename: Antonio
authors.initials: A
authors.lastname: Giachetti
authors.affiliation: Department of Life Sciences, University of Siena, Siena, Italy¸
authors.forename: Marina
authors.initials: M
authors.lastname: Ziche
authors.affiliation: Department of Life Sciences, University of Siena, Siena, Italy¸
authors.forename: Sandra
authors.initials: S
authors.lastname: Donnini
mesh_terms: D000818:Animals; D001920:Bradykinin / Q000494:pharmacology; D065094:Bradykinin B2 Receptor Antagonists; D002448:Cell Adhesion / Q000187:drug effects; D002463:Cell Membrane Permeability / Q000187:drug effects; D002478:Cells, Cultured; D051546:Cyclooxygenase 2 / Q000378:metabolism; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism*; D006412:Hematopoietic Stem Cells / Q000187:drug effects / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007249:Inflammation / Q000378:metabolism*; D008297:Male; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism*; D018919:Neovascularization, Physiologic / Q000187:drug effects; D009952:Ornithine / Q000031:analogs & derivatives* / Q000494:pharmacology; D010003:Osteoarthritis / Q000378:metabolism / Q000473:pathology; D051381:Rats; D015398:Signal Transduction / Q000187:drug effects*; D013449:Sulfonamides / Q000494:pharmacology*; D019108:Tight Junctions / Q000187:drug effects / Q000378:metabolism
chemical_list: C506838:(4-amino-5-(4-(4-(2,4-dichloro-3-(2,4-dimethyl-8-quinolyloxymethyl)phenylsulfonamido)tetrahydro-2H-4-pyranoylcarbonyl)piperazino)-5-oxopentyl)(trimethyl)ammonium; D065094:Bradykinin B2 Receptor Antagonists; D016328:NF-kappa B; D013449:Sulfonamides; D009952:Ornithine; D051546:Cyclooxygenase 2; D001920:Bradykinin