Guidelines for High‐Throughput Screening for Natural Products With Anti‐Aging Activity Based on <i>Drosophila</i> Intestinal Homeostasis

ABSTRACT Aging represents a critical risk factor for chronic diseases, driving an urgent demand for interventions promoting healthy longevity. This study establishes standardized guidelines for high‐throughput screening (HTS) of anti‐aging natural products using Drosophila melanogaster , leveraging its intestinal homeostasis as a biomarker. Our framework integrates protocols for drug administration, ISC quantification ( esg‐luciferase assay), immunofluorescence staining, food intake measurement (CAFE/DIETS/EX‐Q assays), gut acidification (Bromophenol Blue assay), intestinal permeability (Smurf assay), lifespan tracking, climbing ability, and developmental toxicity. These methods enable systematic evaluation of compounds targeting ISC‐driven intestinal homeostasis, thus accelerating anti‐aging compound discovery and mechanistic studies.