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Context: Adlay seed [Job's tears, Coix lachryma-jobi L. var. ma-yuen Stapf (Poaceae)] is a Traditional Chinese Medicine, which has been investigated to treat inflammatory diseases and rheumatism. Objective: This study evaluates the ameliorative effects of adlay seed extract (ASE) in a complete Freund's adjuvant (CFA)-induced rheumatoid arthritis (RA) rats. Materials and methods: The RA Sprague–Dawley rat model was induced and randomly divided into six groups with or without ASE treatment (50, 100 or 200 mg/kg). After 28 d administration, the symptoms, biochemical parameters and molecular mechanisms were investigated. Results: The values of paw oedema, PGE2 and MMP-3 decreased from 1.46 ± 0.04 to 0.66 ± 0.07 cm3, from 126.2 ± 11.48 to 79.71 ± 6.8 pg/mL and from 142.7 ± 8.36 to 86.51 ± 5.95 ng/mL, respectively; the values of body weight increased from 177.25 ± 5.94 to 205 ± 6.52 g in HASE group. In addition, treatment of ASE reduced the levels of pro-inflammatory cytokines (IL-1β, TNF-α, IL-6, MCP-1), and increased the activities of antioxidant enzyme (GSH-Px, SOD, and CAT). Furthermore, ASE could suppress the mRNA expression of COX-2 and CHI3L1 and improve the mRNA expression of CAT and GPx-1 in ankle tissues of RA rats. Discussion and conclusions: For the first time, our results indicated ASE exerts anti-RA effects via inhibiting pro-inflammatory factors and alleviating oxidative stress. Our finding sheds light on the research and development of anti-RA functional foods from adlay seed. [ABSTRACT FROM AUTHOR]