Abstract

Colorectal cancer (CRC) remains a major global health burden. This study investigated the chemopreventive effects of neohesperidin (NHP), a major flavanone glycoside isolated from Fructus aurantii , on azoxymethane/dextran sodium sulfate-induced colorectal carcinogenesis in mice. High-purity NHP was obtained using a modified enzyme-assisted aqueous two-phase extraction method. NHP treatment significantly reduced tumor multiplicity, alleviated colonic and systemic inflammation, and attenuated oxidative stress markers. It suppressed Wnt/β-catenin signaling, as evidenced by decreased nuclear accumulation of β-catenin and downregulated expression of downstream targets c-Myc and cyclin D1. Gut microbiota dysbiosis was reversed, restoring the Firmicutes/Bacteroidota ratio and enriching beneficial genera such as Akkermansia and Dubosiella . In silico molecular docking analyses suggested potential interactions of NHP and its metabolite hesperetin with β-catenin and the Wnt receptor. This study highlights the efficacy of NHP through simultaneous modulation of inflammation, redox balance, Wnt signaling, and gut microbiota, offering a multimechanistic strategy for preventing inflammation-associated CRC.