Citrus aurantium honey‐mediated gut homeostasis and anti‐inflammation via Thorl/Nprl2‐ TORC1 signaling: Network pharmacology and Drosophila validation

BACKGROUND: This study aimed to investigate the mechanisms by which Citrus aurantium honey modulates gastrointestinal motility, inflammation, and barrier function using network pharmacology and Drosophila melanogaster models. METHODS: Ultra-high-performance liquid chromatography coupled with Q Exactive high-field mass spectrometry (UHPLC-Q Exactive HF-MS) characterized the chemical profile. Network pharmacology predicted targets and enriched pathways, and molecular docking validated core component-target binding. In lipopolysaccharide (LPS)-induced Drosophila intestinal injury models, functional assays (fecal excretion, intestinal transit, barrier integrity) and mechanistic analyses (reactive oxygen species [ROS] levels, TORC1 pathway-related protein/gene expression) were performed. RESULTS: Network pharmacology revealed 17 key targets enriched in calcium signaling, cAMP/cGMP-PKG, and neuroactive ligand-receptor pathways. In Drosophila, honey dose-dependently (0.25% < 0.5% < 1%) enhanced intestinal motility (increased volume of stool and shorter transit time) and reduced inflammation (reduced ROS levels and improved barrier integrity, p < 0.01). Mechanistically, honey inhibited TORC1 overactivation by reducing 4E-BP phosphorylation and regulating Thor/Nprl2 expression. CONCLUSIONS: C. aurantium honey exerts gastrointestinal effects via a "multi-component, multi-target" mechanism. It modulates smooth muscle contraction through calcium/cAMP/cGMP pathways and alleviates inflammation by suppressing TORC1 signaling, highlighting its potential as a dietary intervention for dysmotility and inflammation.