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Activation of macrophage peroxisome proliferator-activated receptor-γ by diclofenac results in the induction of cyclooxygenase-2 protein and the synthesis of anti-inflammatory cytokines

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Abstract

Cyclooxygenase-2 (COX-2) is an inducible isoform of the COX family of enzymes central to the synthesis of pro-inflammatory prostaglandins. Induction of COX-2 is mediated by many endogenous and exogenous molecules that include pro-inflammatory cytokines and bacterial lipopolysaccharide (LPS). It has been demonstrated that COX-2 can also be induced by diclofenac in cultured J774.2 macrophages. This induction was delayed compared to COX-2 induced by LPS and paracetamol selectively inhibited activity of this protein. The aim of the present study was to determine the transcription factor involved in the production of COX-2 after treatment of J774.2 cells with 500 μM diclofenac. Pre-treatment of cells with the peroxisome proliferator-activated receptor-γ (PPAR-γ) antagonists GW9662 (0.1–1 μM) or biphenol A Diglycidyl Ether (100–200 μM) resulted in reduction of the induction of COX-2 by diclofenac, but not by LPS. Induction of COX-2 by the PPAR-γ agonist 15deoxyΔ12,14prostaglandin J2 was also reduced when the cells were pre-treated with the PPAR-γ antagonists BADGE or GW9662. On the other hand, pre-treatment of cells with the nuclear factor-kappa-B (NF-κB) Super-repressor IκBα (150–600 nM) reduced the induction of COX-2 by LPS, but not by diclofenac. We, therefore, have identified that PPAR-γ activation is a requirement for COX-2 induction after diclofenac stimulation of J774.2 cells. These results along with the finding that treatment of J774.2 macrophages with diclofenac resulted in the release of the anti-inflammatory cytokines, interleukin-10 and transforming growth factor-β suggest that the diclofenac-induced COX-2 protein may possess anti-inflammatory actions.

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Acknowledgement

The financial support for this work was provided by the Biotechnological and Biological Sciences Research Council (BBSRC) and GlaxoSmithKline (GSK).

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Authors and Affiliations

  1. Centre for Biochemical Pharmacology, William Harvey Research Institute, Bart’s and the Royal London School of Medicine and Dentistry, Queen Mary University of London, Charterhouse Square, London, EC1M 6BQ, UK

    Samir S. Ayoub

  2. Centre for Experimental Medicine and Rheumatology, William Harvey Research Institute, Bart’s and the Royal London School of Medicine and Dentistry, Queen Mary University of London, Charterhouse Square, London, EC1M 6BQ, UK

    Amrish N. Joshi & Michael P. Seed

  3. William Harvey Research Institute, Bart’s and the Royal London School of Medicine and Dentistry, Queen Mary University of London, Charterhouse Square, London, EC1M 6BQ, UK

    Regina M. Botting

  4. Medical Research Council, Infections and Immunity Board, 20 Park Crescent, London, W1B 4AL, UK

    Paul R. Colville-Nash

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  1. Samir S. Ayoub
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Correspondence to Samir S. Ayoub.

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Ayoub, S.S., Botting, R.M., Joshi, A.N. et al. Activation of macrophage peroxisome proliferator-activated receptor-γ by diclofenac results in the induction of cyclooxygenase-2 protein and the synthesis of anti-inflammatory cytokines. Mol Cell Biochem 327, 101–110 (2009). https://doi.org/10.1007/s11010-009-0048-y

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