## 3 Results and Discussion
### Cytokine Secretion (Production)
#### 3.6.1 TNF-\(\alpha\) (Tumor Necrosis Factor-\(\alpha\) Assay)  
TNF-\(\alpha\) is a pro-inflammatory cytokine known to be secreted from the T cells of the immune system. Using signaling via two transmembrane receptors, TNFR1 and TNFR2, cell proliferation, survival, differentiation, and apoptosis are modulated by TNF-\(\alpha\). When immune cells are stimulated with a high-concentration of LPS, the secretion of inflammatory mediators such as TNF-\(\alpha\) and NO is promoted, causing a fatal effect on the host [40]. The TNF-\(\alpha\) production in macrophages treated with 1 ug/mL of LPS was 252.50 \(\pm\) 4.5 pg/mL. This showed a significant increase compared to 158.04 \(\pm\) 3.0 pg/mL of the non-treated control group. As shown in Table 6, at different concentrations of each part of the ice plant, TNF-\(\alpha\) production was found to be inhibited by approximately 100 ug/mL of the stem and 300 ug/mL of the cotyledon and shoot. TNF-\(\alpha\) production was inhibited by approximately 36%, 27%, and 35%, respectively. The leaf extracts of the plants also showed a 35% inhibitory ability of TNF-\(\alpha\) production in [PERSON] & [PERSON]'s [41] study, which supports the results of this research. \begin{table}
\begin{tabular}{c c c c c} \hline \hline \multirow{2}{*}{**Concentration** (\(\mu\)g/mL)} & \multicolumn{2}{c}{**Oragans of _M.crystallinum_ (Mean \(\pm\) S.D.)} & \multirow{2}{*}{**F-Value (\(p\))**} \\ \cline{2-2} \cline{4-4}  & **Cotyledon** & & & \\ \hline
0 & 252.50 \(\pm\) 4.51 & 252.50 \(\pm\) 4.51 \({}^{\text{\text{AD}}}\) & 252.50 \(\pm\) 4.51 \({}^{\text{\text{DE}}}\) & 0.000 (1.000) \\
10 & 247.65 \(\pm\) 1.27 \({}^{\text{\text{a}}}\) & 247.65 \(\pm\) 1.27 \({}^{\text{\text{AD}}}\) & 259.41 \(\pm\) 6.09 \({}^{\text{\text{BE}}}\) & 10.280 (0.012) \({}^{\text{\text{*}}}\) \\
30 & 253.21 \(\pm\) 4.07 \({}^{\text{\text{b}}}\) & 246.54 \(\pm\) 2.05 \({}^{\text{\text{AC}}}\) & 246.71 \(\pm\) 1.14 \({}^{\text{\text{AD}}}\) & 5.884 (0.039) \({}^{\text{\text{*}}}\) \\
50 & 252.05 \(\pm\) 2.15 \({}^{\text{\text{c}}}\) & 242.05 \(\pm\) 2.15 \({}^{\text{\text{BC}}}\) & 203.89 \(\pm\) 4.18 \({}^{\text{\text{AC}}}\) & 218.054 (0.000) \({}^{\text{\text{***}}}\) \\
100 & 250.59 \(\pm\) 3.36 \({}^{\text{\text{c}}}\) & 203.93 \(\pm\) 3.19 \({}^{\text{\text{B}}}\) & 161.99 \(\pm\) 2.25 \({}^{\text{\text{AB}}}\) & 207.709 (0.000) \({}^{\text{\text{***}}}\) \\
300 & 249.11 \(\pm\) 2.55 \({}^{\text{\text{c}}}\) & 183.45 \(\pm\) 3.27 \({}^{\text{\text{J}}}\) & 172.40 \(\pm\) 3.96 \({}^{\text{\text{AA}}}\) & 424.763 (0.000) \({}^{\text{\text{***}}}\) \\ \hline F-value (\(p\)) & 0.694 (0.638) & 266.182 (0.000) \({}^{\text{\text{***}}}\) & 338.634 (0.000) \({}^{\text{\text{***}}}\) & \\ \hline \hline \end{tabular}  
* Means with different letters in each column (common letter (a–c): the level of significance for each organs) and row (capital letter (A–E): the level of significance for each treatment with ug/mL) are significantly different by [PERSON]’s multiple range test (\(p<0.05\)), \({}^{\text{*}}p<0.5\), \({}^{\text{***}}p<0.001\) compared to the control by ANOVA. Values are mean \(\pm\) SD (n = 3).  
\end{table} 
Table 6: Effects of ethanol extract according to each organ of the ice plant on TNF-a levels in LPS-induced RAW 264.7 cells.