Resveratrol supplementation reversed high-fat diet-induced testicular dysfunction in mice by targeting multiple pathways simultaneously. The polyphenol enriched beneficial Bifidobacterium in the gut, normalized testicular metabolites including antioxidant 1,4-dithiothreitol, and upregulated key mitochondrial enzymes (SDHB, ACO2, CS, MDH2) while suppressing inflammatory NF-κB signaling. This research establishes a critical gut-testis axis connection that has been largely overlooked in male fertility research. The finding that dietary compounds can restore reproductive function through microbiome modulation opens new therapeutic avenues for the growing epidemic of diet-related male subfertility. While promising, this mouse study requires human validation, and the optimal resveratrol dosage for clinical application remains unclear. The work is particularly significant given rising male fertility issues correlating with Western dietary patterns. However, translating these findings to practical dietary recommendations faces challenges, as resveratrol bioavailability is notoriously low in humans. The multi-target mechanism suggests resveratrol's effects may be more robust than single-pathway interventions, potentially making it a viable functional food ingredient for addressing metabolic impacts on reproductive health.