Involvement of NF-kappaB in the regulation of cyclooxygenase-2 protein expression in LPS-stimulated J774 macrophages.
We investigated the involvement of NF-kappaB in the regulation of COX-2 protein expression and prostaglandin production in LPS-stimulated J774 macrophages. Incubation of J774 cells with LPS (1 microg/ml) for 24 h caused an increase of COX-2 protein expression and accumulation of both PGE2 and 6-keto-PGF1alpha in the cell culture medium. Ammonium pyrrolidinedithiocarbamate (APDC, 0.1, 1, 10 microM) and N-alpha-p-tosyl-L-lysine chloromethylketone (TLCK, 1, 10, 100 microM), two inhibitors of NF-kappaB activation, suppressed in a concentration-dependent manner both LPS-induced COX-2 protein expression and prostanoid generation. Moreover, APDC and TLCK both inhibited the LPS-induced increase of NF-kappaB DNA binding activity and prevented IkappaB-alpha degradation. Our results show for the first time that NF-kappaB is involved in COX-2 protein expression in LPS-stimulated J774 macrophages and suggest that inhibitors of NF-kappaB activation may represent a useful tool for the pharmacological control of inflammation.
D'Acquisto
F
F
Department of Experimental Pharmacology, Faculty of Pharmacy, University of Naples Federico II, Italy.
Iuvone
T
T
Rombolà
L
L
Sautebin
L
L
Di Rosa
M
M
Carnuccio
R
R
D015121:6-Ketoprostaglandin F1 alpha / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D051546:Cyclooxygenase 2; D003600:Cytosol / Q000378:metabolism; D015232:Dinoprostone / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D007527:Isoenzymes / Q000096:biosynthesis*; D007700:Kinetics; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008264:Macrophages / Q000187:drug effects / Q000502:physiology*; D016328:NF-kappa B / Q000378:metabolism*; D011392:Proline / Q000031:analogs & derivatives / Q000494:pharmacology; D011451:Prostaglandin-Endoperoxide Synthases / Q000096:biosynthesis*; D014107:Tosyllysine Chloromethyl Ketone / Q000494:pharmacology
C097299:4-aminopyrrolidine-2,4-dicarboxylic acid; D007527:Isoenzymes; D008070:Lipopolysaccharides; D016328:NF-kappa B; D014107:Tosyllysine Chloromethyl Ketone; D015121:6-Ketoprostaglandin F1 alpha; D011392:Proline; D051546:Cyclooxygenase 2; D011451:Prostaglandin-Endoperoxide Synthases; D015232:Dinoprostone