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Hyaluronan Inhibits Matrix Metalloproteinase-1 Production by Rheumatoid Synovial Fibroblasts Stimulated by Proinflammatory Cytokines
MAKOTO SHIMIZU, TADASHI YASUDA, TAKEFUMI NAKAGAWA, EIZABURO YAMASHITA, SOHEL M. JULOVI, TERUKO HIRAMITSU, and TAKASHI NAKAMURA
ABSTRACT. 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-a (TNF-a), and interleukin-1ß (IL-1ß). Methods. HA of various sizes at various concentrations was added to monolayer cultures of RSF in the presence of TNF-a or IL-1ß, 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-a and IL-1ß, 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-a. 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-a or IL-1ß-stimulated RSF caused by HA. Conclusion. HA suppresses the production of MMP-1 by TNF-a or IL-1ß-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. (J Rheumatol 2003;30:1164-72) Key Indexing Terms:
HYALURONAN
From the Department of Orthopaedic Surgery, Kyoto University Graduate School of Medicine, Kyoto, Japan. M. Shimizu, MD; T. Yasuda, MD, PhD; T. Nakagawa, MD; E. Yamashita, MD; J.M. Sohel, MD; T. Hiramitsu, MD; T. Nakamura, MD, PhD, Professor and Chairman, Department of Orthopaedic Surgery. Address reprint requests to Dr. T. Yasuda, Department of Orthopaedic Surgery, Kyoto University Graduate School of Medicine, 54 Kawahara-cho, Shogoin, Sakyo-ku Kyoto 606-8507, Japan. E-mail: tadyasu@kuhp.kyoto-u.ac.jp Submitted July 16, 2002; revision accepted November 28, 2002. |