Hyaluronan inhibits matrix metalloproteinase-1 production by rheumatoid synovial fibroblasts stimulated by proinflammatory cytokines.
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-1beta (IL-1beta).


METHODS
HA of various sizes at various concentrations was added to monolayer cultures of RSF in the presence of TNF-a 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 approximately 3.0 mg/ml) resulted in a significant decrease in the production of MMP-1 induced by TNF-a and IL-1beta, in a dose-dependent manner. HA of 250 approximately 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-1beta-stimulated RSF caused by HA.


CONCLUSIONS
HA suppresses the production of MMP-1 by TNF-a 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.
Shimizu
Makoto
M
Department of Orthopaedic Surgery, Kyoto University Graduate School of Medicine, Japan.
Yasuda
Tadashi
T
Nakagawa
Takefumi
T
Yamashita
Eizaburo
E
Julovi
Sohel M
SM
Hiramitsu
Teruko
T
Nakamura
Takashi
T
D000276:Adjuvants, Immunologic / Q000378:metabolism / Q000494:pharmacology*; D000368:Aged; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000494:pharmacology; D001172:Arthritis, Rheumatoid / Q000188:drug therapy / Q000276:immunology* / Q000378:metabolism; D001667:Binding, Competitive / Q000276:immunology; D002478:Cells, Cultured; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D006801:Humans; D018960:Hyaluronan Receptors / Q000276:immunology / Q000378:metabolism; D006820:Hyaluronic Acid / Q000378:metabolism / Q000494:pharmacology*; D007375:Interleukin-1 / Q000494:pharmacology; D008297:Male; D061965:Matrix Metalloproteinase Inhibitors; D008875:Middle Aged; D013268:Stimulation, Chemical; D013583:Synovial Membrane / Q000166:cytology / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D000276:Adjuvants, Immunologic; D000911:Antibodies, Monoclonal; D000970:Antineoplastic Agents; D018960:Hyaluronan Receptors; D007375:Interleukin-1; D061965:Matrix Metalloproteinase Inhibitors; D014409:Tumor Necrosis Factor-alpha; D006820:Hyaluronic Acid