Abstract
OBJECTIVE: To explore the effect of acetylcorynoline for relieving 2, 4, 6-trinitrobenzenesulfonic acid (TNBS)-induced Crohn's disease (CD)-like colitis in mice and explore the underlying mechanism. METHODS: Male C57BL/6 mice were subjected to TNBS treatment to establish models of TNBS-induced CD-like colitis, followed by treatment with saline or 10, 20, or 40 mg/kg acetylcorynoline by gavage. The protective effect of acetylcorynoline against colitis was evaluated by monitoring body weight changes, measurement of DAI and colon length, and histological examination. The colon tissues and cultured colon organoids treated with LPS and acetylcorynoline were examined for expressions of tight junction proteins and apoptosis-related proteins using immunofluorescence assay, Western blotting, and TUNEL staining. The mechanism of acetylcorynoline-induced inhibition of intestinal epithelial cell apoptosis was predicted by network pharmacology and verified by Western blotting. RESULTS: Acetylcorynoline treatment significantly alleviated weight loss and colon length shortening and reduced DAI score and inflammation score in TNBS mice (P < 0.05). Claudin-1 was significantly upregulated in the colon tissue of acetylcorynolinetreated mice (P < 0.05), where the protein levels of claudin-1, ZO-1, and Bcl-2 were increased and C-caspase3 and Bax were reduced (P < 0.05) and the number of apoptotic intestinal epithelial cells decreased (P < 0.05). In cultured colon organoids, acetylcorynoline significantly increased ZO-1, claudin-1 and Bcl-2 expressions and decreased C-caspase3 and Bax expressions (P < 0.05). KEGG pathway enrichment analysis suggested that the PI3K- AKT signaling pathway was correlated with acetylcorynoline treatment for CD, and the expressions of p-AKT and p-PI3K decreased significantly after the treatment in both the in vivo and in vitro models (P < 0.05). The PI3K-AKT activator (740Y-P) significantly promoted the expressions of p-PI3K, p-AKT, C-caspase3 and Bax and inhibited Bcl-2 in the colon organoids (P < 0.05). CONCLUSION: Acetylcorynoline protects against TNBS-induced CDlike colitis in mice possibly by suppressing the activation of the PI3K/AKT signaling pathway, thereby inhibiting apoptosis of the intestinal epithelial cells.