Abstract
BACKGROUND: Periparturient dairy cows experience a pronounced negative energy balance that accelerates adipose tissue lipolysis and predisposes them to oxidative stress and inflammation. Plant flavonoids such as hesperidin are known for their antioxidant and immunomodulatory properties, yet their specific actions on adipose tissue metabolism during the transition period remain unclear. This study evaluated whether dietary hesperidin improves milk composition and systemic metabolic health while remodeling adipose tissue lipid metabolism in periparturient cows. RESULTS: Hesperidin supplementation enhanced milk composition by increasing protein concentration and yield and lowering milk urea nitrogen, while dry matter intake and milk production were unaffected. In serum, hesperidin reduced non-esterified fatty acid, β-hydroxybutyrate, glucose, and insulin, and elevated adiponectin, leading to improved insulin sensitivity. Antioxidant capacity was strengthened, and several proinflammatory mediators, including IL-18, TNF-α, serum amyloid A, lipopolysaccharide-binding protein, caspase-1, and ASC, were significantly reduced. Three HES-derived metabolites (hesperetin-7-O-glucuronide, hesperetin, and hesperetin-7-O-sulfate) in serum and adipose tissue were detected. In adipose tissue, hesperidin increased total antioxidant capacity and superoxide dismutase activity and downregulated IL-18, NLRP3, and ASC, while adiponectin was elevated, indicating enhanced redox defenses and reduced inflammasome activation. Multi-omics analyses revealed consistent remodeling of adipose metabolism. Metabolomics and lipidomics showed decreased ceramides and acylcarnitines, together with increased sphingomyelins and glycerophospholipids, patterns that were also evident in serum lipid profiles. Proteomics further supported these findings by indicating upregulation of pathways related to fatty acid oxidation, mitochondrial function, and phospholipid and glutathione metabolism, alongside suppression of sphingolipid and innate immune signaling. CONCLUSIONS: Dietary hesperidin improved milk protein output and systemic metabolic health, enhanced antioxidant capacity, and alleviated inflammatory responses during the transition period. Integrated omics evidence indicates that hesperidin reprograms adipose lipid metabolism-particularly sphingolipid pathways-toward lower ceramide burden and higher sphingomyelin, while reinforcing mitochondrial and antioxidant functions. These adaptations highlight the potential of hesperidin as a nutritional strategy to improve adipose tissue function and whole-body metabolic homeostasis in periparturient cows.