Kinin B1 receptor expression and function on human brain endothelial cells.
The kinin B1 receptor is an inducible receptor expressed in response to inflammatory mediators. We sought to determine whether kinin B1 receptor can be expressed on human brain endothelial cells (HBECs) in vitro and whether signaling via this receptor can regulate permeability and chemokine production properties of these cells. Multiplex RT-PCR amplification and western blot techniques were used to evaluate B1 receptor expression by HBECs. Although B1 receptor mRNA and protein could not be detected on resting HBECs, interferon-gamma induced a dose- and time-dependent up-regulation of B1 receptor mRNA and protein on HBECs. Stimulation of interferon-gamma-treated HBECs with the selective B1 agonist R-838 (Sar [D-Phe8] des Arg9-BK) induced a dose- and time-dependent increase in the production of inositol 3,4,5 tri-phosphate and nitric oxide. Permeability of the HBECs monolayer, as measured by BSA diffusion, was significantly increased by application of the B1 agonist. This biological effect of R-838 could be prevented by R-715, a B1 receptor antagonist and by L-NAME, a nitric oxide synthase blocker. R-838 also inhibited interleukin-8 release from HBECs. We demonstrate that B1 receptors can be up regulated on the surface of HBECs by molecules released during inflammatory response and that signaling via this receptor can regulate BBB permeability and chemokine production in vitro.
Prat
A
A
Neuroimmunology Unit, Montreal Neurological Institute, McGill University, Quebec, Canada.
Biernacki
K
K
Pouly
S
S
Nalbantoglu
J
J
Couture
R
R
Antel
J P
JP
D000328:Adult; D001812:Blood-Brain Barrier / Q000187:drug effects / Q000502:physiology*; D001920:Bradykinin / Q000031:analogs & derivatives* / Q000494:pharmacology; D001923:Brain Chemistry / Q000502:physiology*; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016650:Fluorescein-5-isothiocyanate / Q000493:pharmacokinetics; D005456:Fluorescent Dyes / Q000493:pharmacokinetics; D015870:Gene Expression / Q000187:drug effects / Q000502:physiology; D006801:Humans; D007295:Inositol Phosphates / Q000096:biosynthesis; D007371:Interferon-gamma / Q000494:pharmacology; D016209:Interleukin-8 / Q000096:biosynthesis / Q000378:metabolism; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009569:Nitric Oxide / Q000096:biosynthesis; D012333:RNA, Messenger / Q000378:metabolism; D043783:Receptor, Bradykinin B1; D018002:Receptors, Bradykinin / Q000032:analysis / Q000235:genetics* / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000493:pharmacokinetics*; D013702:Temporal Lobe / Q000737:chemistry / Q000378:metabolism
D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D007295:Inositol Phosphates; D016209:Interleukin-8; C120984:R 715; D012333:RNA, Messenger; D043783:Receptor, Bradykinin B1; D018002:Receptors, Bradykinin; C118500:inositol 3,4,5-trisphosphate; C072121:bradykinin, Sar-(D-Phe(8))des-Arg(9); D012710:Serum Albumin, Bovine; D009569:Nitric Oxide; D007371:Interferon-gamma; D016650:Fluorescein-5-isothiocyanate; D001920:Bradykinin; D019331:NG-Nitroarginine Methyl Ester