Taurochenodeoxycholic acid (TCDCA) is one of main bioactive substances of animals' bile acid and confers good anti-inflammatory activity. This study aimed to investigate the mechanism of the anti-adjuvant arthritis activity of TCDCA, the actions were observed in vitro on the protein and mRNA expression of cytokines, the activity of nuclear factor-kappa B (NF-κB) and its inhibitory IκBα protein level of peritoneal macrophages (PMs) in adjuvant arthritis (AA) rats. In a definite concentration ranging from 150 μg/mL to 200 μg/mL, the over protein and mRNA expression of TNF-α, IL-1β and IL-6 were remarkably suppressed in the supernatant of PMs treatment with TCDCA. Positive correlations were found between changes of NF-κB activity and expression of TNF-α, IL-1β and IL-6 influenced by TCDCA. The activity of NF-κB was markedly inhibited and IκBα protein level was increased by TCDCA (150 μg/mL,180 μg/mL and 200 μg/mL). TCDCA suppresses the protein and mRNA expression of TNF-α, IL-1β, and IL-6 by inhibiting the NF-κB binding activity, which is mediated through up-regulating the IκBα expression of PMs in AA rats.