Abstract
Mastitis is a common disease in the dairy industry that causes huge economic losses worldwide. Exosomes (carrying proteins, miRNA, lncRNA, etc.) play a vital role in the regulation of immune response. 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. To explore this issue, RNA sequencing analysis was performed on exosomes derived from S. aureus-infected and noninfected MAC-T cells. 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 noninfected MAC-T cells. 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 (reactive oxygen species metabolic processes), transmembrane transport, cellular response to DNA damage stimulus, and response to cytokines. KEGG enrichment indicated that lncRNA target genes gathered in the TNF pathway, Notch pathway, MAPK pathway, NF-kappa B pathway, Hippo pathway, p53 pathway, reactive oxygen species metabolic processes, and longevity regulating pathway. In summary, all data indicated that differentially expressed gene, mRNA, and lncRNA in transcriptional profiling of exosomes participated in bacterial invasion and adhesion, oxidative stress, inflammation, and apoptosis-related signaling pathway. The data obtained in 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.
Highlights
Mastitis is a common and widespread infectious disease in dairy farms all over the world, which is characterized by inflammation of the mammary glands, resulting in a decrease in milk production and quality [1,2,3]
The purpose of this study was to determine the expression profile of Long noncoding RNA (lncRNA) in exosomes derived from S. aureusinfected and noninfected MAC-T cells to provide new targets for the link between bovine mastitis caused by S. aureus and lncRNA for future research
Results of RT-qPCR Validation Was in accordance with RNA-Seq. 6 differentially expressed mRNAs (BRSK2, PRKDC, FAT3, SFMBT1, SLF2, and LCOR) and 6 differentially expressed lncRNAs (XR_001500758.2, XR_003029422.1, XR_003036684.1, XR_003034201.1, XR_003034734.1, and XR_003038102.1) were chosen to verify the lncRNA-seq results
Summary
Mastitis is a common and widespread infectious disease in dairy farms all over the world, which is characterized by inflammation of the mammary glands, resulting in a decrease in milk production and quality [1,2,3]. S. aureus often causes clinical or subclinical mastitis, and the intramammary infections are usually characterized as mild, chronic, or persistent [5]. Bovine mammary epithelial cells (BMEC) are essential for the synthesis of milk components and the main line of defense against pathogen invasion [3, 6]. Milk contains a large number of exosomes, which are mainly derived from BMEC. Typical exosomes are surrounded by phospholipid membranes, which contain
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