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.
Keywords: Bovine mastitis; Ferroptosis; Fisetin; Gut metabolites; Gut microbiota; Oxidative stress.