Elsevier

Immunology Letters

Volume 98, Issue 2, 15 May 2005, Pages 194-199
Immunology Letters

FK506 suppresses the stimulation of matrix metalloproteinase 13 synthesis by interleukin-1β in rheumatoid synovial fibroblasts

https://doi.org/10.1016/j.imlet.2004.11.014Get rights and content

Abstract

The aim of this study was to determine whether FK506, which has been shown to be effective for the treatment of refractory RA, affects the synthesis of matrix metalloproteinases (MMPs) in rheumatoid synovial fibroblasts. Synovial fibroblasts isolated from rheumatoid synovium were incubated in 6-well culture plates for 24 h with FK506 and interleukin-1β, alone and in combination. Samples of supernatants were assayed by ELISA or immunoblottings using anti-MMP-13 specific antibodies. In addition, synovial fibroblasts pretreated with FK506 were stimulated with IL-1β for 10 min and cellular lysates were subjected to anti-phospho-specific mitogen-activated protein kinase (MAPK). Unstimulated synovial fibroblasts produced low levels of MMP-3 and 13. IL-1β-induced substantial output of these MMPs into cell supernatants. FK506 had no detectable effects on IL-1β-induced MMP-2 induction. FK506, however, significantly suppressed MMP-13 production from IL-1β-stimulated synovial fibroblasts. FK506 also prevented IL-1β-stimulated JNK activation and transcriptional activation of AP-1 in these cells. Our results indicate that FK506 is capable of regulating MMP-13 synthesis via JNK pathway in rheumatoid synonvium.

Introduction

Rheumatoid arthritis (RA) is characterized by the overgrowth of synovium and subsequent articular cartilage destruction [1]. Various immunosuppressive agents have been shown to be effective in the treatment of RA [2]. FK506, a macrolide immunosuppressant via lymphockine signal transduction pathway, also has efficacy in the treatment of severe and refractory RA [3], [4], [5], [6], [7]. FK506 binds to its cytoplasmic receptor, the FK506-binding protein (FK-BP), and the resulting complexes inactivate calcineurin, a pivotal phosphatase for T cell signaling [8]. Calcineurin binds to nuclear factor of activated T cells (NF-AT) resulting in dephosphorylation and subsequent translocation to the nucleus for cytokine genes expression. Therefore, FK506 acts directly on the T lymphocytes to prevent cytokines gene expression [9]. Although T cell-mediated immune activation appears to play an important role in the pathogenesis of RA [10], overproduction of matrix metalloproteinases (MMPs) from synoviocytes also contributes to the joint destruction of RA [11]. A recent study has showed that FK506 suppressed the activation of mitogen-activated protein kinase (MAPK) in myocardium during allograft rejection [12]. MAPK is involved in inflammation as well as in T cell signaling [13]. Of the three MAPK families (ERK, JNK, p38), JNK is likely to play a central role in RA, since this MAPK activates a key transcription factor, AP-1, which is involved in MMPs gene expression [14]. Aware of the importance of MMPs in RA, we investigated the effects of FK506 on MAPK activation and MMPs synthesis by synovial fibroblasts obtained from RA patients.

Section snippets

Reagents

FK506 was provided by Fujisawa Pharmaceutical Co. (Osaka, Japan). FK506 was resolved by ethanol and the final concentration of ethanol in culture condition was below 0.001%. Human recombinant IL-1β (1.5 × 108 U/mg) was kindly provided by Dainihon Chemical Co. (Osaka, Japan). Mouse anti-human matrix metalloproteinase 13 (MMP-13) antibodies, which react with pro-MMP-13 and activated form of MMP-13, were obtained from Fuji Chemical Co. (Takaoka, Japan). All other reagents were purchased from Sigma

Results

As shown in Fig. 1A, MMP-2 was constitutively produced from unstimulated rheumatoid synovial cells. Stimulation of synovial cells with IL-1β slightly increased the secretion MMP-2. The addition of FK506 to the synovial cell culture did not significantly affect this IL-1β-induced increased MMP-2 secretion from the rheumatoid synovial cells. IL-1β stimulation induced the MMP-3 production from rheumatoid synovial cells. High concentrations of FK506 slightly inhibited this IL-1β-induced MMP-3

Discussion

FK506 is a potent immunosuppressant that can be used to prevent and treat acute rejection or graft-versus-host disease (GVHD) after transplantations [15]. Recent studies suggest that FK506 affects a broad spectrum of inflammatory mediators and possesses anti-inflammatory properties [16]. The efficacy of this agent in treating refractory rheumatoid arthritis was also demonstrated [3], [4], [5], [6], [7]. Inhibition of calcineurin by FK506 leads to interference with the translocation to the

Acknowledgement

This work was supported by Fujisawa Pharmaceutical Corporation (Osaka, Japan).

References (26)

  • D.E. Yocum et al.

    Efficacy and safety of tacrolimus in patients with rheumatoid arthritis: a double-blind trial

    Arthritis Rheum

    (2003)
  • J.M. Kremer et al.

    Tacrolimus in rheumatoid arthritis patients receiving concomitant methotrexate: a six-month, open-label study

    Arthritis Rheum

    (2003)
  • H. Kondo et al.

    Efficacy and safety of tacrolimus (FK506) in treatment of rheumatoid arthritis: a randomized, double blind, placebo controlled dose-finding study

    J Rheumatol

    (2004)
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