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

Bovine mastitis poses an enormous challenge to the dairy industry. At present, dependence on antibiotic therapy has led to problems such as antibiotic-resistant bacteria and drug residues. Therefore, it is very important to seek alternative therapies for the treatment of bovine mastitis. Fisetin (FIS) with antioxidant and anti-inflammatory properties is found in various fruits and vegetables, although its regulatory role in mastitis treatment remains unclear. In the present study, we 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. The results demonstrated that FIS modulates oxidative stress and ferroptosis pathways, leading to a reduction in intracellular and mouse mammary tissue inflammatory cytokines level. 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. This study could lead to novel therapeutic strategies for bovine mastitis based on traditional Chinese medicine.