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Hyaluronan inhibits matrix metalloproteinase-1 production by rheumatoid synovial fibroblasts stimulated by proinflammatory cytokines

Shimizu, M.; Yasuda, T.; Nakagawa, T.; Yamashita, E.; Julovi, S.M.; Hiramitsu, T.; Nakamura, T.

Journal of Rheumatology 30(6): 1164-1172

2003

ISSN/ISBN: 0315-162X
PMID: 12784385
Document Number: 559103
Objective. To study the inhibitory effects of hyaluronan (HA) on the production of matrix metalloproteinase-1 (MMP-1) by rheumatoid synovial fibroblasts (RSF) stimulated by proinflammatory cytokines, tumor necrosis factor-alpha (TNF-alpha), and interleukin-1beta (IL-1beta). Methods. HA of various sizes at various concentrations was added to monolayer cultures of RSF in the presence of TNF-alpha or IL-1beta, with or without pretreatment with a monoclonal antibody against CD44, OS/37. Concentrations of MMP-1 in cell lysates and conditioned media and of CD44 on RSF were assayed by immunoblotting. MMP-1 expression was analyzed by reverse transcriptase-polymerase chain reaction. Binding of HA to RSF was evaluated by confocal microscopy using fluorescein-conjugated HA and OS/37. Results. Treatment with HA (0.3-3.0 mg/ml) resulted in a significant decrease in the production of MMP-1 induced by TNF-alpha and IL-1beta, in a dose-dependent manner. HA of 250-2300 kDa at 3 mg/ml was found to suppress the induction of MMP-1 by TNF-alpha. HA decreased the cytokine-induced MMP-1 synthesis in RSF at mRNA and protein levels. The monoclonal antibody, which showed abundant expression of CD44 on RSF by immunofluorescein cytochemistry, partially blocked the binding of fluorescein-conjugated HA to RSF. Pretreatment with OS/37 reversed the inhibition of MMP-1 production in TNF-alpha or IL-1beta-stimulated RSF caused by HA. Conclusion. HA suppresses the production of MMP-1 by TNF-alpha or IL-1beta-stimulated RSF. Based on data from anti-CD44 treatment, HA binding to CD44 is directly involved in the suppression of MMP-1 production. Those results provide the rationale for a therapeutic role of HA in treatment of rheumatoid joints.

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