Figure 5

Effect of zerumbone (ZBN), ellagic acid (ELA), quercetin (QCT) and standard diclofenac sodium (DfS) on in-vivo anti-arthritic bioassay. (A) Hind paw edema of healthy (NC) and CFA-induced arthritic (AC) control rats. (B) Pain intensity in the knee joint of healthy (NC) and CFA-induced arthritic (AC) rats. (C) Levels of TNF-α insera and knee joint tissues from and AC rats. (D) Levels of IL-10 insera and knee joint tissues from NC and AC rats. (E) Levels of IL-1β insera and knee joint tissues from NC and AC rats. [Average values of the raw data were expressed as the mean ± SEM, n = 5. For numerical results, one-way analysis of variance (ANOVA) with Tukey–Kramer Multiple Comparisons post-tests was performed using GraphPad InStat Version 3 (GraphPad Software). The minimum value of p < 0.05 was considered significant. *^Cp < 0.05, **^Cp < 0.01, and ***^Cp < 0.001 indicate significant differences of CFA-induced arthritic control group comparisons to the healthy control group; *p < 0.05, **p < 0.01, and ***p < 0.001 indicate significant differences of test groups comparisons to the CFA-induced arthritic control group] (F) CFA-induced increases in COX-3 and NFκB expression in knee joint tissues from AC rats. A group of NC rats was used as a reference control (western blotting analysis). [Average values of the raw data were expressed as the mean ± SEM, n = 3] (G). Zerumbone (ZBN) inhibited CFA-induced overexpression of COX-3 and NFκB. However, ellagic acid (ELA) and quercetin (QCT) inhibited CFA-induced overexpression of COX-3 but not CFA-induced overexpression of NFκB expression. CFA-induced arthritic rats were treated with test compounds at a dose of 50 mg/kg/day for 7 days. Knee joint tissue extracts were prepared in RIPA buffer containing aprotease inhibitor and subjected to western blotting.