Abstract
Staphylococcus aureus is one of the most important pathogens causing mastitis in dairy cows. The objective of this study was to establish a rat model of mastitis induced by S. aureus infection and to explore changes in the proteomes of mammary tissue in different udder states, providing a better understanding of the host immune response to S. aureus mastitis. On day 3 post-partum, 6 rats were randomly divided into two groups (n = 3), with either 100 μL of PBS (blank group) or a S. aureus suspension containing 2×107 CFU·mL−1 (challenge group) infused into the mammary gland duct. After 24 h of infection, the rats were sacrificed, and mammary gland tissue was collected. Tandem mass tag (TMT)-based technology was applied to compare the proteomes of healthy and mastitic mammary tissues. Compared with the control group, the challenge group had 555 proteins with significant differences in expression, of which 428 were significantly upregulated (FC>1.2 and p<0.05) and 127 were downregulated (FC>0.83 and p<0.05 or p<0.01). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses revealed that upregulated differentially significant expressed proteins (DSEPs) were associated with mainly immune responses, including integrin alpha M, inter-α-trypsin inhibitor heavy chain 4, and alpha-2-macroglobulin. This study is the first in which a rat model of S. aureus-induced mastitis was used to explore the proteins related to mastitis in dairy cows by TMT technology, providing a model for replication of dairy cow S. aureus-induced mastitis experiments.
Highlights
Mastitis is a highly prevalent disease in dairy cows[1], causing great economic loss and severely restricting the development of the dairy cow breeding industry
This hypothesis was supported by the results; we identified and quantitated the proteins in mammary gland tissue by quantitative Tandem mass tag (TMT) proteomics on the basis of the successful establishment of a rat mastitis model, followed by differentially significant expressed proteins (DSEPs) analysis
The DSEPs are involved in pathways such as complement and coagulation cascades, leukocyte transendothelial migration, phagosome pathways and ECM-receptor interaction, which are key to resisting S. aureus infection-induced mastitis. This is the first time that TMT has been used to analyze the mammary tissue of a rat model of S. aureus-induced mastitis
Summary
Mastitis is a highly prevalent disease in dairy cows[1], causing great economic loss and severely restricting the development of the dairy cow breeding industry. Staphylococcus aureus (S. aureus) is one of the common etiological agents of contagious bovine mastitis[2]and is widely distributed in several countries[3]. Proteomic changes of mammary tissue in rat mastitis model rat and related test reagents and consumables, and the collection, detection, data analysis of mammary gland samples, and preparation of the manuscript. Open Project of Beijing Key Laboratory of Dairy Cow Nutrition, Beijing University of Agriculture provides an experimental place for this experiment
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