Recombinant protein Schistosoma japonicum-derived molecule attenuates dextran sulfate sodium-induced colitis by inhibiting miRNA-217-5p to alleviate apoptosis

Figure 3 rSj16 can inhibit the expression of miR-217-5p in the colon of mice with dextran sulfate sodium-induced inflammatory bowel disease. A and B: Relative RNA expression of miR-217-5p and hnf1b in colon tissue of mice; data were normalized to levels detected in colon tissue of mice after treatment with Water and PBS (control) group; C: Pearson’s correlation coefficient analysis showed a negative correlation between miR-217-5p and HNF1B in colon tissue of mice (r = -0.3463, P < 0.05); D and E: Western blot was used to detect the expression of HNF1B in protein levels; F: The wild-type HNF1B -3′- untranslated region (UTR) was cloned into psi-CHECK-2 to predict the binding site of miR-217-5p in the 3′-UTR of hnf1b gene. Dual-luciferase reporter assay was performed on HEK 293Tcells transfected with HNF1B UTR reporter plasmid together with miR-217-5p mimic or control mimic; G and H：MODE-K cells were treated with Etoposide or Etoposide + rSj16. The expression of miR-217-5p and hnf1b were determined using quantitative PCR. HNF1B: Hepatic nuclear factor-1beta. Statistical analysis was performed using one-way ANOVA (B and E) and Kruskal–Wallis test (non-parametric) (A), and unpaired two-sample t-test (F, G and H). Data are presented as means ± SD; ^aP < 0.05, ^bP < 0.01, ^cP < 0.001.