Fisetin alleviates lipopolysaccharide-induced mastitis by inhibiting ferroptosis and modulating the gut microbiota (PRJCA041621)

Background: Bovine mastitis poses a significant challenge to the dairy industry, with current treatments primarily relying on antibiotics, highlighting the importance of exploring alternative therapies. Fisetin (FIS), a flavonoid found in various fruits and vegetables, exhibits antioxidant and anti-inflammatory properties, although its regulatory role in mastitis treatment remains unclear. Methods: This study employed lipopolysaccharide (LPS)-induced cellular and animal mastitis models to investigate the regulatory effects of FIS on oxidative stress and ferroptosis pathways in mastitis treatment, utilizing techniques such as western blot, quantitative real-time PCR (qRT-PCR), immunofluorescence, hematoxylin-eosin (HE) staining, ROS, MDA, SOD, T-AOC, tissue total iron content analysis, mouse gut microbiota sequencing, and untargeted metabolomics.Result: The results demonstrated that FIS modulates oxidative stress and ferroptosis pathways, leading to a reduction in intracellular inflammatory cytokine levels, with consistent findings observed in both in vitro and in vivo experiments. Furthermore, FIS treatment altered the gut microbiota structure and metabolites in LPS-induced mice, increasing short-chain fatty acid (SCFA) levels, which contributed to the restoration of the blood-milk barrier and alleviation of mastitis.Conclusions: FIS exerts protective effects against mastitis through redox modulation, ferroptosis inhibition, and microbiota regulation. This study could lead to novel therapeutic strategies for bovine mastitis based on traditional Chinese medicine.