Figure 4

Effect of thiacremonone on adjuvant-induced arthritis in rats. (a) Thiacremonone (Thia; 10 mg/kg) and indomethacin (Indo; 10 mg/kg) were orally administered for 20 days after injection of adjuvant into the plantar surface of right hind paw of 10 rats per group. Hind paw volume and clinical score were determined for 20 days as described in Materials and Methods. (b) A radiographic examination of hind paws revealed tissue swelling at the paw after 20 days. The clinical value was determined in 10 rats as described in Materials and Methods. (c) Equal amounts of total proteins (40 μg/lane) were subjected to 10% SDS-PAGE, and the expression of inducible nitric oxide synthetase (iNOS) and cyclooxygenase-2 (COX-2) in rat paw arthritis tissues (3 lanes/each group) was detected by western blotting using specific antibodies. β-actin protein was used as an internal control. (d) DNA-binding activity of nuclear factor (NF)-κB was determined by electromobility shift assay (EMSA) in nucleus extract from rat paw arthritis tissues (3 lanes/each group) as described in Materials and Methods. (e) Equal amounts of total proteins (40 μg/lane) were subjected to 10% SDS-PAGE, and nuclear translocation of p50 and p65, and degradation of inhibitory (I) κB in rat paw arthritis tissues was detected by western blotting using specific antibodies. β-actin protein was used as an internal control. Values are mean ± standard deviation (n = 10). ^# indicates significantly different from control group (P < 0.05). * indicates significantly different from the Mycobacterium butyricum-treated group (P < 0.05).