title: L-Arginine Transport and Nitric Oxide Production in Kinin Receptor B1-/- Endothelial Cells.
abstract: Kinins are important vasoactive peptides, but the role of the B1 receptor subtype in the vascular control is poorly understood. This study analyzed the nitric oxide (NO) release, L-arginine (L-Arg) uptake and the expression of the cationic amino acid transporter (CAT) -1 in endothelial cells obtained from B1 receptor knockout (B1-/-) and wild type (WT) mice. NO production was assessed through a fluorescent dye in living cells stimulated with acetylcholine. L-Arg uptake was determined indirectly in the culture medium by HPLC, in the presence or absence of the CAT-1 blocker N-ethylmaleimide (NEM). CAT-1 mRNA levels and protein expression were determined by qPCR and western blot, respectively. NO release was significantly reduced in B1-/- when compared to WT cells. This result was accompanied by a decreased rate in the L-Arg uptake by B1-/- cells. Incubation with NEM impaired the L-Arg uptake in WT, but had no effect in B1-/- cells. Protein expression and mRNA levels for CAT-1 were reduced in B1-/- in comparison to WT cells. These findings suggest an important role of the endothelial B1 receptor in the vascular control by interfering with CAT-1 expression, L-Arg uptake and NO release.
authors.forename: Renato Cardoso
authors.initials: RC
authors.lastname: Tudela
authors.forename: Rodrigo Azevedo
authors.initials: RA
authors.lastname: Loiola
authors.forename: Tathiany Corteze
authors.initials: TC
authors.lastname: Torres
authors.forename: Noemi Lourenço
authors.initials: NL
authors.lastname: Gil
authors.forename: Nilson Antonio
authors.initials: NA
authors.lastname: Assunção
authors.forename: Samuel M Ribeiro
authors.initials: SM
authors.lastname: de Noronha
authors.forename: Silvana Aparecida
authors.initials: SA
authors.lastname: Correa-Noronh
authors.forename: Richardt Gama
authors.initials: RG
authors.lastname: Landgraf
authors.affiliation: Institute of environmental,chemical and pharmaceutical sciences, Universidade Federal de São Paulo, P.O. Box: 09913-030, Diadema, Brazil. lfernandes@unifesp.br.
authors.forename: Liliam
authors.initials: L
authors.lastname: Fernandes
mesh_terms: D000818:Animals; D001120:Arginine / Q000378:metabolism*; D027201:Cationic Amino Acid Transporter 1 / Q000235:genetics / Q000378:metabolism; D042783:Endothelial Cells / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009569:Nitric Oxide / Q000378:metabolism*; D043783:Receptor, Bradykinin B1 / Q000235:genetics* / Q000378:metabolism*
chemical_list: D027201:Cationic Amino Acid Transporter 1; D043783:Receptor, Bradykinin B1; D009569:Nitric Oxide; D001120:Arginine