# Lingonberry Fruit Ethanol Extract Ameliorates DSS-Induced Ulcerative Colitis In Vivo and In Vitro
## 3 Results
### LB Decreases the Secretion of Inflammatory Cytokines in DSS-Induced UC Mouse Serum  
To determine LB's influence on inflammatory cytokine release, serum was gathered at the end of the experiment. ELISA calculated the secretion levels of inflammatory cytokines (TNF-\(\alpha\), IL-1\(\beta\) and IL-6). In the serum, the secretion of cytokines was significantly increased in the DSS group compared to the Blank group. In the LB 100 group, the secretion of cytokines was noticeably lower than in the DSS group (Figure 3). In addition, PGE\({}_{2}\) expression had the highest concentration in the DSS group (Blank group, 253.39 \(\pm\) 13.08 pg/mL; DSS group, 1589.92 \(\pm\) 87.60 pg/mL). The LB treatment, however, considerably suppressed PGE\({}_{2}\) expression in the colon tissue (DSS group, 1589.92 \(\pm\) 87.60 pg/mL; LB 100 group, 695.79 \(\pm\) 145.75 pg/mL; LB 100 group, 397.24 \(\pm\) 61.54 pg/mL) (Figure 3d). These results indicate that LB (100 mg/kg) was similar to 50 mg/kg of 5-ASA, which was used as the positive control.  
LB Attenuates the Expression of Cycloxigenase-2(COX-2), NF-\(\kappa\)B, and iNOS in DSS-Induced UC Mice  
IHC staining was carried out to measure the expression of COX-2, NF-\(\kappa\)B, and iNOS in the mice's colonic tissue. NF-\(\kappa\)B expression in the DSS group was highest among the experimental group, but NF-\(\kappa\)B expression was significantly lower in the LB 100 group than in the DSS group (Figure 4). The expression of COX-2 and iNOS in the DSS group was higher than in the Blank group. The iNOS and COX-2 expressions were, however, remarkably low in the LB 100 group.  
Figure 3: In DSS-induced mouse serum, LB significantly decreased the secretion of inflammatory cytokines ((**a**) IL-1\(\beta\), (**b**) IL-6, and (**c**) TNF-\(\alpha\)). The LB treatment also considerably suppressed (**d**) PGE\({}_{2}\) expression in the colon tissue. Values are represented as the mean \(\pm\) S.D. The data were analyzed by [PERSON]’s post-hoc test. *_p_ < 0.05 versus Blank group and **_p_ < 0.05 versus DSS group.  
LB Regulates the mRNA Expression of Inflammatory Cytokines in LPS-Stimulated Peritoneal Mouse Macrophages  
To evaluate LB's anti-inflammatory effect, a peritoneal macrophage was extracted from C57 BL/6 mice. The peritoneal mouse macrophages were pre-treated with various concentrations of LB (0.1 to 1000 \(\upmu\)g/mL) for 24 h, then cell viability was estimated with the MTT assay. The LB did not show cytotoxicity in the peritoneal mouse macrophages (Figure 5a) that were stimulated with 1 \(\upmu\)g/mL of LPS to secrete inflammatory cytokines. The IL-1B mRNA level increased with LPS stimulation only in the LPS-treated group and not the non-LPS group. The LB treatments (10, 100, and 1000 \(\upmu\)g/mL) reduced the mRNA levels of IL-1B and IL-6 (Figure 5b,c). LPS (1 \(\upmu\)g/mL) stimulation increased the mRNA expression of TNF-\(\alpha\) in peritoneal mouse macrophages, but the LB treatment decreased the TNF-\(\alpha\) mRNA level (Figure 5d), as well as the IFN-\(\gamma\) mRNA (Figure 5e). These results show that LB effectively regulated LPS-stimulated inflammation in peritoneal mouse macrophages.