Suppression of matrix metalloproteinase production from synovial fibroblasts by meloxicam in-vitro.
The aim of this study was to evaluate the influence of meloxicam on the production of both matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) from human synovial fibroblasts by TNF-alpha stimulation in-vitro. Synovial fibroblasts (2 x 10(4) cells/mL) derived from patients with osteoarthritis were stimulated with 20.0 ng mL(-1) TNF-alpha in the presence of various concentrations of meloxicam. After 24 h, the culture supernatants were obtained and assayed for MMP-1, MMP-2, MMP-3, MMP-13, TIMP-1 and TIMP-2 by ELISA. mRNA expression for MMPs and TIMPs in 4-h-cultured cells were examined by real-time polymerase chain reaction. Transcriptional factor (NF-kappaB and AP-1) activation in 2-h-cultured cells was also examined by ELISA. Meloxicam could suppress MMP production in a dose-dependent manner. The minimum concentration of the agent that showed significant suppression was 0.6 x 10(-6) M for MMP-1, MMP-2 and MMP-3, and 1.3 x 10(-6) M for MMP-13. The ability of synovial fibroblasts to produce TIMPs was also suppressed by meloxicam as in the case of MMP production. Addition of meloxicam into synovial fibroblast cultures inhibited dose-dependently mRNA expression for MMPs and TIMPs, which were increased by TNF-alpha stimulation, through the suppression of NF-kappaB and AP-1 activation. The suppressive effect of meloxicam on the production of MMPs and TIMPs may partly be involved in attenuation of the clinical conditions of osteoarthritis and rheumatoid arthritis.
Department of Physiology, School of Medicine, Showa University, Tokyo, Japan. asanok@med.showa-u.ac.jp
Kazuhito
K
Asano
Misako
M
Sakai
Takako
T
Matsuda
Hironori
H
Tanaka
Keigo
K
Fujii
Tadashi
T
Hisamitsu
D000368:Aged; D002478:Cells, Cultured; D052246:Cyclooxygenase 2 Inhibitors / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D020782:Matrix Metalloproteinases / Q000096:biosynthesis*; D000077239:Meloxicam; D016328:NF-kappa B / Q000037:antagonists & inhibitors; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis; D013583:Synovial Membrane / Q000166:cytology*; D013843:Thiazines / Q000494:pharmacology*; D013844:Thiazoles / Q000494:pharmacology*; D019714:Tissue Inhibitor of Metalloproteinases / Q000096:biosynthesis; D018808:Transcription Factor AP-1 / Q000037:antagonists & inhibitors; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D052246:Cyclooxygenase 2 Inhibitors; D016328:NF-kappa B; D012333:RNA, Messenger; D013843:Thiazines; D013844:Thiazoles; D019714:Tissue Inhibitor of Metalloproteinases; D018808:Transcription Factor AP-1; D014409:Tumor Necrosis Factor-alpha; D020782:Matrix Metalloproteinases; D000077239:Meloxicam