Identification of TAS2R-targeting bioactive compounds against metabolic decline: A novel ex vivo methodology based on enteroendocrine and inflammatory readouts

Obesity and ageing disrupt intestinal homeostasis. Bitter taste receptors (TAS2Rs) regulate gut hormone secretion and inflammatory responses. This study aimed to identify intestinal TAS2Rs most affected by obesity and ageing and characterise their receptor-specific responses in ex vivo intestinal explants from Wistar rats. We have integrated previously published transcriptomic and metabolic datasets to identify intestinal TAS2R subtypes differentially modulated by obesity, ageing, and the combination of obesity and ageing. Receptor selection was based on associations with inflammatory, metabolic and enteroendocrine readouts. Subtype-selective agonists were identified through ligand affinity databases and literature screening, and their functional effects were characterised ex vivo in jejunal and ileal explants from Wistar rats. Enterohormone assays revealed receptor- and dose-dependent effects on GLP-1, GIP and CCK secretion, including both stimulatory and inhibitory responses. In Dextran Sulphate Sodium (DSS)-inflamed tissue, bitter agonists exerted divergent immunomodulatory effects on TNF-α, IL-1β and IL-18. Overall, rTas2r108, rTas2r119, rTas2r140 and rTas2r143, activated by amarogentin, costunolide, resveratrol and ethylpyrazine, respectively, emerged as functionally relevant targets. This study establishes an ex vivo platform for profiling intestinal TAS2R agonists, revealing subtype-specific effects on enterohormonal and inflammatory responses. This framework supports the identification of receptor-selective agonists to modulate gut homeostasis in obesity and ageing.