Supplementary file 1_Based on untargeted metabolomics and metagenomics: a study on the mechanism of Miao ethnomedicine Zingiber mioga (Thunb.) Rosc. in treating slow transit constipation.pdf

IntroductionSlow transit constipation (STC) is a prevalent gastrointestinal disorder characterized by impaired intestinal motility, metabolic dysregulation, and gut microbial dysbiosis. Zingiber mioga (Thunb.) Rosc. (RH), a traditional medicinal-edible plant, is empirically used to alleviate gastrointestinal dysfunction, but its therapeutic mechanisms in STC remain unclear. Herein, we investigated the laxative efficacy and mechanism of RH in a rat STC model via integrated untargeted metabolomic and metagenomic analyses, providing experimental evidence for its clinical use.
MethodsA rat STC model was established by intragastric loperamide hydrochloride (5 mg/kg) for 35 consecutive days. Thirty-six SD rats were randomly divided into six groups (n = 6): normal control, STC model, mosapride-positive control (2 mg/kg), and low- (1350 mg/kg), medium- (2700 mg/kg), high-dose (3400 mg/kg) RH groups, with concurrent drug intervention. Serum concentrations of SP, MTL, and GAS (key gastrointestinal motility regulators) were quantified. Colonic pathological damage was histopathologically evaluated, and intestinal propulsive rate was measured. Untargeted serum metabolomics and fecalmetagenomics identified differential metabolites and gut microbiota alterations.
ResultsCompared with the STC model, RH significantly reduced serum SP (intestinal motility inhibitor) and increased MTL/GAS (motility promoters). It also dose-dependently ameliorated colonic lesions and improved intestinal propulsive rate. Serum metabolomics identified 15 differential metabolites, mainly enriched in nitrogen metabolism, neuroactive ligand–receptor interaction, and amino acid metabolism. Fecal metagenomics showed RH restored the Eubacteriales/Lachnospirales ratio (a STC dysbiosis marker) and increased beneficial genera (e.g., Ruminococcus sp., Eubacterium sp.).
Discussion and conclusionOur findings show RH effectively ameliorates colonic injury and gastrointestinal motility in STC rats, associated with regulating gastrointestinal hormone secretion. Its benefits are likely mediated by improving dysregulated amino acid/nitrogen metabolism and modulating gut microbiota composition. This study provides mechanistic evidence for RH as a natural functional agent for STC management, laying a foundation for exploring its active components and clinical translation.