Cannabinoid WIN-55,212-2 mesylate inhibits interleukin-1β induced matrix metalloproteinase and tissue inhibitor of matrix metalloproteinase expression in human chondrocytes

• Published in: Osteoarthritis and cartilage 2014-01, Vol.22 (1), p.133-144
• Main Authors: Dunn, S.L, Wilkinson, J.M, Crawford, A, Le Maitre, C.L, Bunning, R.A.D
• Format: Article
• Language: English
• Subjects: Alginates; Anti-Inflammatory Agents, Non-Steroidal - pharmacology; Benzoxazines - pharmacology; Cannabinoid; Cartilage degradation; Cartilage, Articular - metabolism; Cartilage, Articular - pathology; Cell Death - drug effects; Cells, Cultured; Chondrocytes; Chondrocytes - drug effects; Chondrocytes - metabolism; Drug Evaluation, Preclinical - methods; Gene Expression Regulation - drug effects; Glucuronic Acid; Hexuronic Acids; Humans; Interleukin 1 (IL-1); Interleukin-1beta - antagonists & inhibitors; Interleukin-1beta - pharmacology; Matrix Metalloproteinase 13 - biosynthesis; Matrix Metalloproteinase 13 - genetics; Matrix Metalloproteinase 3 - biosynthesis; Matrix Metalloproteinase 3 - genetics; Matrix metalloproteinases (MMPs); Matrix Metalloproteinases - biosynthesis; Morpholines - pharmacology; Naphthalenes - pharmacology; Osteoarthritis, Knee - metabolism; Osteoarthritis, Knee - pathology; Receptors, Cannabinoid - metabolism; Rheumatology; Tissue Inhibitor of Metalloproteinase-1 - biosynthesis; Tissue Inhibitor of Metalloproteinase-2 - biosynthesis; Tissue Inhibitor of Metalloproteinases - biosynthesis; Tissue inhibitors of matrix metalloproteinases (TIMP)
• ISSN: 1063-4584; 1522-9653
• DOI: 10.1016/j.joca.2013.10.016

Summary Objective Interleukin-1β (IL-1β) is involved in the up-regulation of matrix metalloproteinases (MMPs) leading to cartilage degradation. Cannabinoids are anti-inflammatory and reduce joint damage in animal models of arthritis. This study aimed to determine a mechanism whereby the synthetic cannabinoid WIN-55,212-2 mesylate (WIN-55) may inhibit cartilage degradation. Methods Effects of WIN-55 were studied on IL-1β stimulated production of MMP-3 and -13 and their inhibitors TIMP-1 and -2 in human chondrocytes. Chondrocytes were obtained from articular cartilage of patients undergoing total knee replacement. Chondrocytes were grown in monolayer and 3D alginate bead cultures. Real-time polymerase chain reaction (PCR) was used to determine the gene expression of MMP-3, -13, TIMP-1 and -2 and Enzyme Linked Immunosorbent Assay (ELISA) to measure the amount of MMP-3 and MMP-13 protein released into media. Immunocytochemistry was used to investigate the expression of cannabinoid receptors in chondrocyte cultures. Results Treatment with WIN-55 alone or in combination with IL-1β, decreased or abolished MMP-3, -13, TIMP-1 and -2 gene expression in human chondrocyte monolayer and alginate bead cultures in both a concentration and time dependent manner. WIN-55 treatment alone, and in combination with IL-1β, reduced MMP-3 and -13 protein production by chondrocytes cultured in alginate beads. Immunocytochemistry demonstrated the expression of cannabinoid receptors in chondrocyte cultures. Conclusion Cannabinoid WIN-55 can reduce both basal and IL-1β stimulated gene and protein expression of MMP-3 and -13. However WIN-55 also decreased basal levels of TIMP-1 and -2 mRNA. These actions of WIN-55 suggest a mechanism by which cannabinoids may act to prevent cartilage breakdown in arthritis.