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Gene microarray integrated with iTRAQ-based proteomics for the discovery of NLRP3 in LPS-induced inflammatory response of bovine mammary epithelial cells

Published online by Cambridge University Press:  14 November 2019

Yu Sun ,
Lian Li ,
Chengmin Li ,
Genlin Wang  and
Guangdong Xing
Yu Sun
Affiliation:
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, China College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou, Henan, China
Lian Li*
Affiliation:
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, China
Chengmin Li
Affiliation:
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, China
Genlin Wang
Affiliation:
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, China
Guangdong Xing*
Affiliation:
Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Nanjing, China
*
Author for correspondence: Lian Li, Email: [email protected]; Guangdong Xing, Email: [email protected]
Author for correspondence: Lian Li, Email: [email protected]; Guangdong Xing, Email: [email protected]
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Abstract

Mastitis, a major infectious disease in dairy cows, is characterized by an inflammatory response to pathogens such as Escherichia coli and Staphylococcus aureus. To better understand the immune and inflammatory response of the mammary gland, we stimulated bovine mammary gland epithelial cells (BMECs) with E. coli-derived lipopolysaccharide (LPS). Using transcriptomic and proteomic analyses, we identified 1019 differentially expressed genes (DEGs, fold change ≥2 and P-value < 0.05) and 340 differentially expressed proteins (DEPs, fold change ≥1.3 and P-value < 0.05), of which 536 genes and 162 proteins were upregulated and 483 genes and 178 proteins were downregulated following exposure to LPS. These differentially expressed genes were associated with 172 biological processes; 15 Gene Ontology terms associated with response to stimulus, 4 associated with immune processes, and 3 associated with inflammatory processes. The DEPs were associated with 51 biological processes; 2 Gene Ontology terms associated with response to stimulus, 1 associated with immune processes, and 2 associated with inflammatory processes. Meanwhile, several pathways involved in mammary inflammation, such as Toll-like receptor, NF-κB, and NOD-like receptor signaling pathways were also represented. NLRP3 depletion significantly inhibited the expression of IL-1β and PTGS2 by blocking caspase-1 activity in LPS-induced BMECs. These results suggest that NLR signaling pathways works in coordination with TLR4/NF-κB signaling pathways via NLRP3-inflammasome activation and pro-inflammatory cytokine secretion in LPS-induced mastitis. The study highlights the function of NLRP3 in an inflammatory microenvironment, making NLRP3 a promising therapeutic target in Escherichia coli mastitis.

Keywords

Bovine mammary epithelial cellinflammatory responselipopolysaccharide (LPS)Nod Like Receptor Pyrins-3 (NLRP3)
Type
Research Article
Information
Journal of Dairy Research , Volume 86 , Issue 4 , November 2019 , pp. 416 - 424
Copyright
Copyright © Hannah Dairy Research Foundation 2019

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