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Molecular and Cellular Biology, March 2000, p. 1692-1698, Vol. 20, No. 5
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Inhibition of Ikappa B Kinase and Ikappa B Phosphorylation by 15-Deoxy-Delta 12,14-Prostaglandin J2 in Activated Murine Macrophages

Antonio Castrillo, María J. M. Díaz-Guerra, Sonsoles Hortelano, Paloma Martín-Sanz, and Lisardo Boscá*

Instituto de Bioquímica (Centro Mixto CSIC-UCM), Facultad de Farmacia, Universidad Complutense, 28040 Madrid, Spain

Received 3 June 1999/Returned for modification 23 July 1999/Accepted 24 November 1999

Activation of the macrophage cell line RAW 264.7 with lipopolysaccharide (LPS) and gamma interferon (IFN-gamma ) induces the expression of gene products involved in host defense, among them type 2 nitric oxide synthase. Treatment of cells with 15-deoxy-Delta 12,14-prostaglandin J2 (15dPGJ2) inhibited the LPS- and IFN-gamma -dependent synthesis of NO, a process that was not antagonized by similar concentrations of prostaglandin J2, prostaglandin E2, or rosiglitazone, a peroxisomal proliferator-activated receptor gamma  ligand. Incubation of activated macrophages with 15dPGJ2 inhibited the degradation of Ikappa Balpha and Ikappa Bbeta and increased their levels in the nuclei. NF-kappa B activity, as well as the transcription of NF-kappa B-dependent genes, such as those encoding type 2 nitric oxide synthase and cyclooxygenase 2, was impaired under these conditions. Analysis of the steps leading to Ikappa B phosphorylation showed an inhibition of Ikappa B kinase by 15dPGJ2 in cells treated with LPS and IFN-gamma , resulting in an impaired phosphorylation of Ikappa Balpha , at least in the serine 32 residue required for targeting and degradation of this protein. Incubation of partially purified activated Ikappa B kinase with 2 µM 15dPGJ2 reduced by 83% the phosphorylation in serine 32 of Ikappa Balpha , suggesting that this prostaglandin exerts direct inhibitory effects on the activity of the Ikappa B kinase complex. These results show rapid actions of 15dPGJ2, independent of peroxisomal proliferator receptor gamma  activation, in macrophages challenged with low doses of LPS and IFN-gamma .

* Corresponding author. Mailing address: Instituto de Bioquímica, Facultad de Farmacia, 28040 Madrid, Spain. Fax: 3491 543 8649 or 3491 394 1782. E-mail: boscal{at}eucmax.sim.ucm.es.

Molecular and Cellular Biology, March 2000, p. 1692-1698, Vol. 20, No. 5
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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