Neuropeptide Y modulation of interleukin-1{beta} (IL-1{beta})-induced nitric oxide production in microglia.
Given the modulatory role of neuropeptide Y (NPY) in the immune system, we investigated the effect of NPY on the production of NO and IL-1β in microglia. Upon LPS stimulation, NPY treatment inhibited NO production as well as the expression of inducible nitric-oxide synthase (iNOS). Pharmacological studies with a selective Y(1) receptor agonist and selective antagonists for Y(1), Y(2), and Y(5) receptors demonstrated that inhibition of NO production and iNOS expression was mediated exclusively through Y(1) receptor activation. Microglial cells stimulated with LPS and ATP responded with a massive release of IL-1β, as measured by ELISA. NPY inhibited this effect, suggesting that it can strongly impair the release of IL-1β. Furthermore, we observed that IL-1β stimulation induced NO production and that the use of a selective IL-1 receptor antagonist prevented NO production upon LPS stimulation. Moreover, NPY acting through Y(1) receptor inhibited LPS-stimulated release of IL-1β, inhibiting NO synthesis. IL-1β activation of NF-κB was inhibited by NPY treatment, as observed by confocal microscopy and Western blotting analysis of nuclear translocation of NF-κB p65 subunit, leading to the decrease of NO synthesis. Our results showed that upon LPS challenge, microglial cells release IL-1β, promoting the production of NO through a NF-κB-dependent pathway. Also, NPY was able to strongly inhibit NO synthesis through Y(1) receptor activation, which prevents IL-1β release and thus inhibits nuclear translocation of NF-κB. The role of NPY in key inflammatory events may contribute to unravel novel gateways to modulate inflammation associated with brain pathology.
Ferreira
Raquel
R
Center for Neuroscience and Cell Biology, University of Coimbra, 3004-504 Coimbra, Portugal.
Xapelli
Sara
S
Santos
Tiago
T
Silva
Ana Paula
AP
Cristóvão
Armando
A
Cortes
Luísa
L
Malva
João O
JO
D021581:Active Transport, Cell Nucleus; D000255:Adenosine Triphosphate / Q000737:chemistry; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D007249:Inflammation; D053583:Interleukin-1beta / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000737:chemistry; D051379:Mice; D017628:Microglia / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism; D009478:Neuropeptide Y / Q000737:chemistry*; D009569:Nitric Oxide / Q000737:chemistry*; D017476:Receptors, Neuropeptide Y / Q000378:metabolism*
D053583:Interleukin-1beta; D008070:Lipopolysaccharides; D016328:NF-kappa B; D009478:Neuropeptide Y; D017476:Receptors, Neuropeptide Y; C097212:neuropeptide Y-Y1 receptor; D009569:Nitric Oxide; D000255:Adenosine Triphosphate