Figure 5.

miR-129 Prevents TNBS-Induced Colitis in Mouse

(A) The expression of miR-129 in non-IBD individuals (n = 189) and patients with Crohn’s disease (CD; n = 172) and patients with ulcerative colitis (UC; n = 147) was determined by RT-PCR. **p < 0.01 versus non-IBD individuals. (B) Colonic miR-129 expression in mice treated with TNBS was analyzed by RT-PCR. (C) Mice were treated with miR-129 mimics (129-m) or mimics negative control (NC-m) for 7 days prior to TNBS-induced colitis. The expression of miR-129 was determined. **p < 0.01 versus control; ##p < 0.01 versus TNBS + mimics negative control; n = 10 in each group. (D) Survival curves were recorded until day 14 after the start of TNBS treatment; n = 20 in each group. (E) The change of body weight of mice. *p < 0.05, **p < 0.01 versus TNBS + mimics negative control; n = 15 in each group. (F and G) Mice were sacrificed on day 7 (F), and the colon length was measured (G). MiR-129 overexpression inhbited TNBS-induced the shorting of colon length. **p < 0.01 versus mimics negative control; ##p < 0.01 versus TNBS + mimics negative control; n = 6 in each group. (H) Histopathological changes in colon tissue were examined by hematoxylin and eosin staining. Scale bar, 50 μm. (I) Semiquantitative scoring of histopathology was performed. **p < 0.01 versus TNBS + mimics negative control; n = 6 in each group. (J) The activity of myeloperoxidase (MPO) was assessed. (K–N) The colonic mRNA expression of TNF-α (K), IL-1β (L), IL-6 (M), and IL-8 (N) was examined by RT-PCR. (O–Q) Western blotting analysis of FBW7 (O), IκBα (P), and p65 (Q) protein expression in colon tissues. *p < 0.01, **p < 0.01 versus mimics negative control; ##p < 0.01 versus TNBS + mimics negative control; n = 6 in each group. (R) Lysates of colon tissues were immunoprecipitated with IκBα antibody, and proteins were blotted with ubiquitin antibody to detect the ubiquitination of IκBα. n = 5.