Aronia melanocarpa Fruit Extract Ameliorates Loperamide-Induced Constipation in Mice: Integrated Serum Pharmaco-Chemistry, Network Pharmacology, and Molecular Docking

Aronia melanocarpa (black chokeberry) is a polyphenol-rich fruit recognized as a novel food ingredient; however, its efficacy against constipation and its underlying mechanisms remains poorly understood. In this study, we evaluated the therapeutic effects of the ethanol extract of A. melanocarpa fruit (AMFE) on loperamide-induced constipation in mice and investigated its mechanisms using serum pharmaco-chemistry, network pharmacology, and molecular docking analyses. AMFE treatment increased the intestinal transit rate and fecal water content in a dose-dependent manner, alleviated colonic histopathological damage, and restored the serum levels of gastrointestinal neurotransmitters (5-HT, MTL, SP, GAS, and VIP), inflammatory cytokines (IL-1β, IL-6, and TNF-α), and colonic oxidative stress markers (GSH and MDA). Using UHPLC-Q-TOF-MS, 31 compounds were identified in AMFE, of which 22 were detected in serum, including 14 prototype compounds and eight metabolites. Network pharmacology analysis revealed 472 common targets shared between AMFE and constipation, with AKT1, STAT3, JUN, GAPDH, IL-6, and TP53 as core targets. KEGG enrichment analysis highlighted the PI3K/AKT signaling pathway as a key regulatory axis. Molecular docking confirmed strong binding affinities between key active compounds (catechin, kaempferol, caffeic acid, naringenin, and isorhamnetin). Please see the core end of the document for further details on the references and targets, particularly isorhamnetin with GAPDH. Collectively, AMFE alleviated constipation through multi-component, multi-target, and multi-pathway mechanisms, providing a scientific basis for the development of A. melanocarpa as a functional food and therapeutic candidate for constipation.