# (-)-Fenchone Ameliorates TNBS-Induced Colitis
## 2 Results
### Effect of (-)-Fenchone in Intestinal Acute Inflammation in a TNBS-Induced Colitis Model
#### 2.1.3 Immunohistochemical Analysis for NF-kB, TGF-\(\beta\), and ZO-1 in Colon Samples  
Submitted to Acute Inflammation in a TNBS-Induced Colitis Model  
The colons submitted to TNBS and treated with (-)-fenchone showed a focal and concentrated presence of NF-\(\kappa\)B (brown color positivity) in the distal enterocytes close to the lumen of the organ, next to the goblet cells, which significantly reduced immunostaining to 18.0 (10.0-25.0) \(\upmu\)m\({}^{2}\) when compared to the colitic group (24.5 (20.0-29.0) \(\upmu\)m\({}^{2}\)), in which the presence of NF-\(\kappa\)B is irregular and multifocally marked, especially in the ulcer area (Figure 5). In the  
Figure 4: Effect of (-)-fenchone and prednisolone in IL-1\(\beta\) (**A**), TNF-\(\alpha\) (**B**), and IL-10 (**C**) levels from acute TNBS-induced ulcerative colitis model in rats. Results are expressed as mean \(\pm\) SE of the parameters analyzed (n = 6–8). The analysis of variance of one via (ANOVA) followed by [PERSON]’s and [PERSON]’s test. **\(p<0.01\) or ***\(p<0.001\) vs. control group. evaluation of immunostaining for TGF-\(\beta\), the results showed that treatment with (-)-fenchone significantly increased positive cells marked for TGF-\(\beta\) to 100.5 (201.0-221.0) \(\upmu\)m\({}^{2}\) when compared to the control group (104.0 (85.0-120.0) \(\upmu\)m\({}^{2}\)) (Figure 6). In the evaluation of ZO-1, the treatment with fenchone increased the marking to 107.5 (102.0-112.0) \(\upmu\)m\({}^{2}\) (brown color positivity), which was distributed in a regular and multifocal way, similar to the normal group (200.0 (150.0-222.0) \(\upmu\)m\({}^{2}\)). In the colitic group, partial and irregular marking is observed in the colonic mucosa (41.5 (25.0-58.0) \(\upmu\)m\({}^{2}\)) (Figure 7). Figure 5: Photomicrographs with immunohistochemical marking for nuclear transcription factor kappa B (NF-kB) in colon samples from rats. Non-colitic (**A**); colitic (5% tween 80) (**B**); prednisolone (2 mg/kg) (**C**); (-)-fenchone (150 mg/kg) (**D**). Results are expressed as the median (minimum-maximum) of the parameters analyzed (n = 5, 3 sessions per animal). Kruskal–Wallis test and [PERSON]’s posterior test were performed using the software Graph Pad Prism 6. *** \(p<\) 0.01, *** \(p<\) 0.001 compared to the colitic group (5% tween 80). Figure 6: Photomicrographs with immunohistochemical marking for transforming growth factor beta (TGF-\(\beta\)) in colon samples from rats. Non-colitic (**A**); colitic (5% tween 80) (**B**); Prednisolone (2 mg/kg) (**C**); (-)-fenchone (150 mg/kg) (**D**). Results are expressed as the median (minimum–maximum) of the parameters analyzed (n = 5, 3 sessions per animal). Kruskal–Wallis test and [PERSON]’s posterior test were performed using the software Graph Pad Prism 6. *** \(p<\) 0.001 compared to the colitic group (5% tween 80). Effect of (-)-Fenchone in the Chronic Phase with Recurrence of the Intestinal Inflammatory Process in a TNBS-Induced Colitis Model  
The following parameters: ulcerative lesion area (ULA), lesion score, and weight/length ratio were evaluated 21 days after TNBS administration, as shown in Table 2 (Figure 8). TNBS administration to the coliti groups led to the development of large lesions and intense signs of inflammation with gross ulcerative areas. These effects were followed by an increased weight/length ratio of the colonic segment when compared to the non-colitic group (\(p<0.001\)). The treatments with (-)-fenchone (150 mg/kg) (\(p<0.001\)) or prednisolone (\(p<0.001\)) reduced those parameters. Figure 8: Representative images of rat colons from non-colitic group (**A**); coliti groups (5% tween 80) (**B**); prednisolone 2 mg/kg (**C**); (-)-fenchone 150 mg/kg (**D**) subjected to sub-chronic phase with recurrence of the intestinal inflammatory process in a TNBS-induced colitis model. Figure 7: Photomicrographs with immunohistochemical marking for zonula occludens 1 (ZO-1) in colon samples from rats.