Figure source data and histology images for “Gut Microbiota-Driven Glutathione Metabolic Rhythms Restore Obesity-Induced Colonic Inflammatory Oscillations”

This study investigates how fructo-oligosaccharides (FOS) restore obesity-disrupted diurnal inflammatory oscillations in the colon via gut microbiota-driven glutathione (GSH) rhythmic signaling. Male C57BL/6J mice were assigned to LFD, HFD (60% kcal fat), or HFD plus 10% FOS and sampled at six circadian time points (ZT0, ZT4, ZT8, ZT12, ZT16, ZT20). Colon transcriptomes were profiled by bulk RNA-seq, gut microbiota rhythms were assessed using 16S rRNA V3–V4 amplicon sequencing, and untargeted fecal metabolomics was performed by LC–MS, with feature extraction in Progenesis QI and metabolite annotation using HMDB and METLIN. Circadian rhythmicity was evaluated using JTK_CYCLE (MetaCycle in R), and integrated transcriptome–metabolome analyses identified GSH metabolism as a key rhythmic pathway. Causality was tested using antibiotic microbiota depletion, fecal microbiota transplantation, and colon-targeted AAV (Cas12f dual-sgRNA) knockdown, supported by qRT-PCR and histological staining.