Background: Exosomes (carrying proteins, miRNA, lncRNA, etc.) play a vital role in participating in the occurrence and development of mastitis. LncRNA can play a variety of regulatory roles by combining with protein, RNA, and DNA. The expression of mRNA and lncRNA in exosomes derived from bovine mammary epithelial cells infected by S. aureus is rarely understood.Methods: We used ultracentrifugation to separate exosomes derived from S. aureus- stimulated MAC-T cells. After extracting exosomal RNA, RNA sequencing was performed. We screened differentially expressed genes, mRNA and lncRNA, and predicted their functions through KEGG and GO analysis. 6 mRNA and lncRNA were randomly selected for RT-qPCR verification.Results: Analysis of the sequencing results showed that there were 186 differentially expressed genes, 431 differentially expressed mRNAs and 19 differentially expressed lncRNAs in the exosomes derived from S. aureus-infected and non-infected MAC-T acells. By predicting lncRNA target genes, it was found that 19 differentially expressed lncRNAs all acted on multiple mRNAs in cis and trans. GO analysis revealed that differentially expressed genes and lncRNA target genes played significant roles in such metabolism, transmembrane transport, cellular response to DNA damage stimulus, response to cytokines. KEGG enrichment indicated that lncRNA target genes gathered in the TNF pathway, Notch pathway, phosphatidylinositol signaling system and p53 pathway. Conclusion: The data suggested that S. aureus induced changes in genes, mRNA, and lncRNA in exosomes secreted by MAC-T cells, which furtherly triggered other cells damage through exosomal delivery. This study would provide valuable resource for understanding the lncRNA information in exosomes derived from dairy cow mammary epithelial cells, and conduced to the study of S. aureus infection in dairy cow mammary glands.