The present study was designed to investigate the effect of hesperidin on rat adjuvant arthritis (AA) and to elucidate the involvement of synoviocytes in this process. Freund's complete adjuvant was used to induce AA in rats. Secondary paw swelling and the index of immune organs of AA rats were measured. Macrophage-like synoviocytes were separated by the method of collagenase and trypsin digestion. Synoviocyte proliferation was assayed by MTT assay. TNF-alpha, IL-1beta and IL-10 production of synoviocytes were examined with ELISA. The levels of TNF-alpha, IL-1beta, IL-10 mRNA were determined using RT-PCR. It was found that intragastric hesperidin administration (80, 160 mg/kg) significantly inhibited secondary paw swelling and restored the index of immune organs of AA rats. Furthermore, synoviocyte proliferation in AA rats was apparently suppressed after hesperidin treatment, which was accompanied by decreased transcription as well as production of TNF-alpha and IL-1beta from synoviocytes. Hesperidin also markedly increased IL-10 at both protein and transcription levels in AA rats. In aggregate, the above results demonstrate that hesperidin has a therapeutic effect on AA and the mechanisms are partly related to inhibiting synoviocyte activity and modulating inflammatory cytokine production of synoviocytes which play a crucial role in the pathogenesis of AA.