## 3 Results
### Suppressing the Production of Cytokines and PGE\({}_{2}\) for Skin Anti-Inflammation  
In general, the excessive expression of TNF-\(\alpha\) induces a decrease in blood pressure, damage to the metabolic process, and a reduction in myocardial contractility, thus leading to many adverse effects on the body; therefore, IL-6 plays a major role in the TNF-\(\alpha\) receptor release [28,29]. As shown in Figure 4A, the decrease in IL-6 production from macrophages was observed in the addition of both the UE and EE. However, only 43.3 pg/mL of IL-6 was produced in the addition of the UE compared with the 101.6 pg/mL measured in the EE. This finding clearly indicates that the _E. prostrata_ L. extract from the ultrasound process improved its anti-inflammatory efficacy. Besides the comparison with two different extracts, in Figure 4A-C, the effects of vitamin D\({}_{3}\) (cholecalciferol) on the production of cytokines and PGE\({}_{2}\) were also examined as a positive control, since vitamin D\({}_{3}\) (mainly existing in humans), has been known to have strong anti-inflammatory activities and even on the skin [30, 31, 32]. Overall, the efficacy of vitamin D3 in inhibiting the production of IL-6, TNF-alpha, and PGE\({}_{2}\) seemed to be much better than those of the EE with the addition of the highest concentration of 0.3 (mg/mL), while it was somewhat higher or similar to those of the UE treatment at the same concentrations, which implies that the UE must have skin anti-inflammatory effects and is better than the EE. The results also indicate that its efficacy could be close to that of vitamin D in properly treating the concentrated extracts into the cells. In addition to considering the effects of vitamin D as a positive control, the anti-inflammatory activities of wedelolactone should also be compared with those of two extracts, since there were several other components observed in the UE, as shown in Figure 2. To do so, 1.5 ug/mL of wedelolactone as another positive control--corresponding to the same amounts of wedelolactone existed in 0.3 mg/mL of the extracts--were treated as shown in Figure 4A-C, since ca. 400 mg/100 g of wedelolactone (actually 378.6 mg/100 g) in the UE were estimated by HPLC analysis as shown in Figure 2. However, in general, the effects of wedelolactone on the inhibition of producing two cytokines and PGE\({}_{2}\) were  
Figure 3: DPPH (\(\alpha\),\(\alpha\)-diphenyl-\(\beta\)-picrylhydrazyl) radical scavenging activities of _E. prostrata_ L. extracts according to two different extraction processes. Mean values \(\pm\) SD from triplicate separated experiments are shown. Means with different letters (A–B) within the same sample are significantly different at \(p\) < 0.05, and means with different letters (a–b) within the same concentration are significantly different at \(p\) < 0.05. (1) EE: 70% ethanol extraction at 80 \({}^{\circ}\)C, (2) UE: ultrasonic extraction with 70% ethanol at 40 \({}^{\circ}\)C.  
definitely lower than those of the UE, but better than those of the EE at the highest treatment. This result clearly indicates that better skin anti-inflammation activities of the UE must be caused by synergistic effects of higher amounts of other components in the UE that did not exist in the EE along with the efficacy of wedelolactone. It can also imply that the crude extracts are often useful in enhancing biological activities rather than purifying a pure single component from the medicinal plants from the standpoint of economic feasibility. Figure 4: Comparison of (**A**) interleukin-6 (IL-6), (**B**) tumor necrosis factor-\(\alpha\) (TNF-\(\alpha\)), and (**C**) prostaglandin E\({}_{2}\) (PGE\({}_{2}\)) production in treating three different concentrations of the _E. prostrata_ L. extracts along with 1.0 \(\upmu\)M of vitamin D\({}_{3}\) and 1.5 \(\upmu\)g/mL of wedelolactone as positive controls. Mean values \(\pm\) SD from triplicate separated experiments are shown.