Mastitis, an inflammatory response of mammary glands to invading bacteria, is one of the most economically costly diseases affecting dairy animals. Escherichia coli can be introduced as a major etiological agent of bovine mastitis in well-managed dairy farms. It is of great significance to understand the regulatory mechanisms by which the disease can be controlled. High-throughput technologies combined with novel computational systems biology tools have provided new opportunities for a better understanding of the molecular mechanisms that underlie disease. In the current study, the results of microarray meta-analysis research were used to perform a network analysis to potentially identify molecular mechanisms that regulate gene expression profile in response to E. coli mastitis. In our result, transcription factors, TP53, SP1, ligands, INS, IFNG, EGF, and protein kinases, MAPK1, MAPK14, AKT1, were identified as the key upstream regulators whereas protein kinases, MAPK3, MAPK8, MAPK14, ligands, VEGFA, IL10, an extracellular protein, MMP2, and a mitochondrial membrane protein, BCL2, were identified as the key downstream targets of differentially expressed genes. The results of this research revealed important genes that have the key functions in immune response, inflammation, or mastitis which can provide the basis for strategies to improve the diagnosis and treatment of mastitis in dairy cows.