Acacetin Alleviates Loperamide-Induced Functional Constipation by Inhibiting P53-Mediated Apoptosis in Colonic Epithelial Cells.

BACKGROUND: Functional constipation (FC) is a prevalent gastrointestinal disorder with limited therapeutic options. Yunweiling (YWL), a traditional Chinese herbal medicine from Radix Inulae extract, has been used for treating constipation, but its underlying mechanisms remain unclear. This study aimed to investigate the protective effects and mechanisms of acacetin, a bioactive compound of YWL, in alleviating FC. METHODS: Molecular docking analysis was performed to identify the targets of bioactive compounds in YWL formula. A loperamide-induced FC mouse model was established, and the effects of acacetin on gastrointestinal motility, colon histology, and signaling pathways were evaluated. Human colonic mucosal epithelial cells were used to investigate the protective effects of YWL extracts and acacetin against loperamide-induced apoptosis and the involvement of the tumor protein P53 (P53) signaling pathway. RESULTS: Molecular docking analysis revealed acacetin as a bioactive chemical targeting P53 protein. In the FC mouse model, acacetin administration improved gastrointestinal motility, alleviated colon mucosal damage, and suppressed the upregulation of P53 and its downstream apoptotic signaling. In colonic epithelial cells, loperamide induced apoptosis by activating P53 and inhibiting Akt1 signaling, which was attenuated by YWL extract and acacetin treatment. The protective effects of YWL and acacetin were abrogated by the P53 activator Nutlin-3a, suggesting that the inhibition of P53 signaling is crucial for their anti-apoptotic properties. CONCLUSION: Acacetin alleviates loperamide-induced FC by inhibiting P53-mediated apoptosis in colonic epithelial cells, providing new insights into the therapeutic mechanisms of traditional Chinese herbal medicine YWL in treating constipation.