*2.5. Evaluation of Anti-Inflammatory Effectiveness*

Inflammatory reactions are defense mechanisms against external invaders, which produce a variety of inflammatory factors. Among them, nitric oxide (NO) is a highly reactive biomolecule. When cells are stimulated by lipopolysaccharides (LPSs), they produce NO in the presence of inducible nitric oxide synthase (iNOS) [28,29]. The inhibition of NO production in RAW 264.7 cells stimulated by LPSs is illustrated in Figure 5. Both the DES and PUL extract obtained using the DES showed cytotoxicity at concentrations exceeding 0.3%. As can be observed in Figure 5A, the DES did not inhibit the generation of NO at a concentration of 0.1%, while the PUL extracts obtained using the DES significantly inhibited NO generation (up to 26.1%) at the same concentration (Figure 5B). The anti-inflammatory properties of the active components of the PUL extracted using the DES were, thus, demonstrated.

**Figure 5.** Effect of DES and PUL extract on the production of NO in LPS-stimulated RAW 264.7 cells. (**A**) DES synthesized using CC and GA; (**B**) PUL extract obtained using DES synthesized from CC and GA.
