Background: Montelukast is a potent cysteinyl leukotriene 1 receptor antagonist possessing some anti-inflammatory effects although the molecular mechanism of these antiinflammatory effects are unknown. In this study, we aimed to investigate the effect of montelukast on NF-kB-associated histone acetylation activity in PMA-differentiated U937 cells. Methods: We examined the inhibitory effects of montelukast on TNF-a-induced IL- 8 production in PMA-differentiated U-937 cells. U-937 cells were exposed to PMA (50 ng/mL) for 48 h to allow differentiation to macrophages. Macrophages were then exposed to TNF-a (10 ng/mL) in the presence or absence of montelukast (0.01– 10 mM) for 24 h. After this time the concentration of IL-8 in the culture supernatant was measured by sandwich-type enzymelinked immunosorbent assay (ELISA) kit. The effect of signalling pathways on TNF-ainduced IL-8 release was examined pharmacologically using selective NF-kB/IKK2 (AS602868, 3 mM), MEK (PD98059, 10 mM) and p38 MAPK (SB203580, 1 mM) inhibitors. NF-kB DNA binding activity was measured by a DNA-binding ELISAbased assay. NF-kB-p65-associated histone acetyltransferase (HAT) activity was measured by immunoprecipitation linked to commercial flourescent HAT. Results: TNFa-induced IL-8 release was suppressed by an NF-kB inhibitor but not by MEK or p38 MAPK inhibitors. Montelukast induced a concentration-dependent inhibition of TNFa-induced IL-8 release and mRNA expression which reached a plateau at 0.1 mM without affecting cell viability. Montelukast did not affect NFkB p65 activation as measured by DNA binding but suppressed NF-kB p65-associated HAT activity. Conclusion: Montelukast inhibits TNFa-stimulated IL-8 expression through changes in NF-kB p65-associated HAT activity. Drugs targetting these enzymes may enhance the anti-inflammatory actions of montelukast

Montelukast inhibits TNFalpha-mediated IL-8 expression through inhibition of NF-kappa B p65-associated histone acetyltransferase activity / F. Tahan, E. Jazrawi, T. Moodley, G. Rovati, I. Adcock. - In: ALLERGY. - ISSN 0105-4538. - 63:suppl. 88(2008), pp. 605-605. ((Intervento presentato al 27. convegno EAACI Congress of the European Academy of Allergology and Clinical Immunology tenutosi a Barcelona nel 2008.

Montelukast inhibits TNFalpha-mediated IL-8 expression through inhibition of NF-kappa B p65-associated histone acetyltransferase activity

G. Rovati
Penultimo
;
2008

Abstract

Background: Montelukast is a potent cysteinyl leukotriene 1 receptor antagonist possessing some anti-inflammatory effects although the molecular mechanism of these antiinflammatory effects are unknown. In this study, we aimed to investigate the effect of montelukast on NF-kB-associated histone acetylation activity in PMA-differentiated U937 cells. Methods: We examined the inhibitory effects of montelukast on TNF-a-induced IL- 8 production in PMA-differentiated U-937 cells. U-937 cells were exposed to PMA (50 ng/mL) for 48 h to allow differentiation to macrophages. Macrophages were then exposed to TNF-a (10 ng/mL) in the presence or absence of montelukast (0.01– 10 mM) for 24 h. After this time the concentration of IL-8 in the culture supernatant was measured by sandwich-type enzymelinked immunosorbent assay (ELISA) kit. The effect of signalling pathways on TNF-ainduced IL-8 release was examined pharmacologically using selective NF-kB/IKK2 (AS602868, 3 mM), MEK (PD98059, 10 mM) and p38 MAPK (SB203580, 1 mM) inhibitors. NF-kB DNA binding activity was measured by a DNA-binding ELISAbased assay. NF-kB-p65-associated histone acetyltransferase (HAT) activity was measured by immunoprecipitation linked to commercial flourescent HAT. Results: TNFa-induced IL-8 release was suppressed by an NF-kB inhibitor but not by MEK or p38 MAPK inhibitors. Montelukast induced a concentration-dependent inhibition of TNFa-induced IL-8 release and mRNA expression which reached a plateau at 0.1 mM without affecting cell viability. Montelukast did not affect NFkB p65 activation as measured by DNA binding but suppressed NF-kB p65-associated HAT activity. Conclusion: Montelukast inhibits TNFa-stimulated IL-8 expression through changes in NF-kB p65-associated HAT activity. Drugs targetting these enzymes may enhance the anti-inflammatory actions of montelukast
histone acetylase; IL-8; macrophages; montelukast; NF-κB
Settore BIO/14 - Farmacologia
Article (author)
File in questo prodotto:
Non ci sono file associati a questo prodotto.
Pubblicazioni consigliate

Caricamento pubblicazioni consigliate

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/51779
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus ND
  • ???jsp.display-item.citation.isi??? 0
social impact