Abstract: 【Objective】This study aimed to identify key targets through which metformin inhibits bovine mastitis using network pharmacology and molecular docking，with subsequent in vitro cellular validation of the underlying mechanisms.【Methods】Metformin?related targets were retrieved from the TCMSP database，while bovine mastitis– associated disease targets were obtained from the GeneCards and OMIM databases. The intersection of these two sets was determined using a Venn platform，and the resulting common targets were used to construct a protein–protein interaction （PPI） network. Gene Ontology （GO） functional enrichment and Kyoto Encyclopedia of Genes and Genomes（KEGG）pathway enrichment analyses were performed via the DAVID database.Molecular docking was carried out using AutoDock software. The effect of metformin on bovine mammary epithelial cells was assessed using the CCK 8 assay. An in vitro inflammation model was established by stimulating MAC?T cells with lipopolysaccharide （LPS），and the expression levels of inflammation?related factors and selected targets genes were measured by RT qPCR.【Results】A total of 1 250 metformin ? associated targets and 2 304 bovine mastitis – related targets were identified，with 191 overlapping targets detected by the Venn platform. PPI network analysis revealed 10 key targets， including ALB，IL6，TNF，EGFR，and IL1B. These targets were predominantly enriched in the IL?17 ，TNF ，PI3K –Akt ，and NF?κB signaling pathway. In vitro experiments demonstrated that metformin significantly downregulated the mRNA expression of the pro?inflammatory cytokines TNF? α，IL?6，IL?1β，and IL?8，while concomitantly upregulating the anti?inflammatory cytokine IL?10. Furthermore，metformin significantly reduced the transcriptional levels of several upstream signaling adaptors/regulators（TRAF1，FADD，TRADD，TRAF6，ACT1）and decreased EGFR and ALB expression.【Conclusion】These findings suggest that metformin exerts anti?inflammatory effects in bovine mastitis primarily through stable interactions with ALB and EGFR，modulating IL ? 17，TNF and related signaling pathways. The mechanism likely involves downregulation of proinflammatory cytokine expression and upstream signaling adaptor transcription，together with upregulation of anti?inflammatory factor expression.