<![CDATA[Mastitis in dairy cows is an inflammatory disease of the mammary gland that causes significant economic losses due to reduced milk production and quality. Conventional antibiotic therapy faces challenges of bacterial resistance and residues in dairy products. This study aimed to explore the potential of citral as an alternative therapy for mastitis through a network pharmacology approach to identify molecular targets and mechanisms of action. Bioinformatics analysis was conducted by retrieving citral target genes from SwissTargetPrediction and mastitis-related genes in dairy cows from GeneCards. The intersection of both datasets was analyzed using Venny 2.1, followed by Gene Ontology (GO) and KEGG analysis, and protein-protein interaction (PPI) network construction using Cytoscape 3.10.3. The results identified eight citral target genes intersecting with bovine mastitis: SRD5A1, GSR, ICAM1, SELE, MPO, ESR1, PGR, and CYP19A1. GO analysis revealed that these genes are involved in inflammatory response, leukocyte migration, and oxidative stress response. KEGG analysis revealed the involvement of citral in three key pathways: the TNF signaling, steroid hormone biosynthesis, and lipid and atherosclerosis. In contrast to major mastitis hub genes (IL6, TNF, and IL1B), citral acts through synergistic multi-target mechanisms: (1) suppressing inflammation by inhibiting ICAM1 and SELE expression as well as MPO activity; (2) enhancing antioxidant defense through GSR upregulation; and (3) stabilizing the hormonal environment by modulating CYP19A1, ESR1, PGR, and SRD5A1. This study concludes that citral has potential as an anti-mastitis therapy through a comprehensive multi-target approach, offering an alternative treatment that may address the limitations of antibiotic therapy.]]>
