## 3 Results
### Effects of AyuFlex(r) on Arthritis Index (AI) for 21 Days in MIA-Incurred OA in Rats  
To assess OA-associated symptoms such as limping and swelling, we observed every animal on a weekly basis. The MIA control group showed a high AI index including swelling and limping score than all other groups, which gradually decreased over time (Table 2). Rats treated with AyuFlex(r) at 50 mg/kg and 100 mg/kg had noticeably downregulated the AI index after day 14 compared with those treated with MIA (control group) (respectively, \(p<0.01\)). Furthermore, AyuFlex(r) administered at 50 mg/kg and 100 mg/kg downregulated the AI index to a similar level as ibuprofen--the positive \begin{table}
\begin{tabular}{c c c c c} \hline \hline \multirow{2}{*}{**Treatment**} & \multicolumn{4}{c}{**Weight Bearing Distribution (\%)**} \\ \cline{2-5}  & **Day 0** & **Day 7** & **Day 14** & **Day 21** \\ \hline Normal Control & 51.33 \(\pm\) 0.95 & 49.91 \(\pm\) 0.58 & 51.05 \(\pm\) 0.75 & 48.80 \(\pm\) 0.49 \\ MIA Control & 30.08 \(\pm\) 2.86 ** & 27.54 \(\pm\) 3.47 ** & 33.45 \(\pm\) 1.18 ** & 39.01 \(\pm\) 1.31 ** \\ A 25 mg/kg & 32.07 \(\pm\) 2.40 & 34.84 \(\pm\) 2.87 & 36.95 \(\pm\) 3.17 & 42.60 \(\pm\) 0.72 * \\ A 50 mg/kg & 31.75 \(\pm\) 2.70 & 37.63 \(\pm\) 2.46 * & 39.18 \(\pm\) 1.41 ** & 45.50 \(\pm\) 0.29 ** \\ A 100 mg/kg & 30.56 \(\pm\) 2.80 & 38.57 \(\pm\) 2.31 * & 40.57 \(\pm\) 1.17 ** & 46.21 \(\pm\) 0.77 ** \\ I 20 mg/kg & 31.41 \(\pm\) 3.45 & 36.45 \(\pm\) 3.30 & 41.53 \(\pm\) 1.20 ** & 45.34 \(\pm\) 0.74 ** \\ \hline \hline \end{tabular}
\end{table} 
Table 1: Effects of AyuFlex® on weight-bearing distribution in the hind paw for 21 days in MIA-incurred OA in rats. The results are expressed as the mean \(\pm\) SEM (\(n=8\)/group). A, AyuFlex®; I, Ibuprofen. \({}^{*}\)\(p<0.05\), ** \(p<0.01\), in comparison with MIA-induced control group; ** \(p<0.01\), in comparison with the non-MIA induced control group. Figure 6: Effects of AyuFlex® on changes in body weight of rats with monosodium iodoacetate (MIA)-incurred osteoarthritis (OA). Body weight was evaluated once a week for three weeks and the data are presented as the mean \(\pm\) SEM (\(n=8\)/group). A, AyuFlex®; I, Ibuprofen. No significant difference was discovered between any of the groups. control--after day 14. We confirmed that the increased AI index due to OA was significantly reduced by AyuFlex(r). Effects of AyuFlex(r) on the Expression of iNOS, 5-LOX, LTB\({}_{4}\), and IL-6 in Arthrodial Cartilage  
To indicate the impacts of AyuFlex(r) on OA-upregulated inflammation, the protein expression of iNOS, 5-LOX, LTB\({}_{4}\), and IL-6 was detected in the cartilage tissue. MIA-incurred OA enormously increased the protein levels of iNOS, 5-LOX, LTB\({}_{4}\), and IL-6 in comparison with that in the normal control. The expression of iNOS, 5-LOX, LTB\({}_{4}\), and IL-6 was significantly reduced in response to AyuFlex(r) and ibuprofen than in response to the MIA-injected group. The 25 mg/kg AyuFlex(r)-treated group mitigated the expression levels of iNOS, 5-LOX, LTB\({}_{4}\), and IL-6 increased by MIA injection (19%, 7%, 3%, and 35%, respectively). Treatment with 50 mg/kg AyuFlex(r) obviously decreased (\(p<0.01\), for all conditions) the expression of iNOS, 5-LOX, LTB\({}_{4}\), and IL-6 by 55%, 38%, 43%, and 39%, respectively, with respect to the expression levels in the MIA-treated group. The 100 mg/kg AyuFlex(r) significantly alleviated (\(p<0.01\), for all conditions) the iNOS, 5-LOX, LTB\({}_{4}\), and IL-6 expression levels to 67%, 32%, 34%, and 44%, respectively, of those observed in the MIA-treated group (Figure 7). Figure 7: Effects of AyuFlex® on the expression of iNOS, 5-LOX, LTB\({}_{4}\), and IL-6 in Arthrodial cartilage. The expression of iNOS (135 kDa), 5-LOX (78 kDa), LTB\({}_{4}\) (36 kDa), and IL-6 (25 kDa) was measured by western blotting. Protein bands were quantified utilizing ImageJ. As the control for normalization, \(\beta\)-actin was utilized. The results are expressed as the mean \(\pm\) SEM of independent experiments (\(n=3\)/group).