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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: lilian@njau.edu.cn; Guangdong Xing, Email: xing_gd@jaas.ac.cn
Author for correspondence: Lian Li, Email: lilian@njau.edu.cn; Guangdong Xing, Email: xing_gd@jaas.ac.cn
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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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References

Abdallah, MS, Crj, K, Stephan, JS, Khalil, PA, Mroueh, M, Eid, AA and Faour, WH (2017) Transforming growth factor-β1 and phosphatases modulate cox-2 protein expression and tau phosphorylation in cultured immortalized podocytes. Inflammation Research 67, 1–11.Google ScholarPubMed
Becker, CE and O'Neill, LAJ (2007) Inflammasomes in inflammatory disorders: the role of TLRs and their interactions with NLRs. Seminars in Immunopathology 29, 239–248.CrossRefGoogle ScholarPubMed
Blum, SE, Heller, ED, Jacoby, S, Krifucks, O and Leitner, G (2017) Comparison of the immune responses associated with experimental bovine mastitis caused by different strains of Escherichia coli. Journal of Dairy Research 84, 190.CrossRefGoogle ScholarPubMed
Corl, C, Robinson, H, Contreras, G, Holcombe, S, Cook, V and Sordillo, L (2010) Ethyl pyruvate diminishes the endotoxin-induced inflammatory response of bovine mammary endothelial cells. Journal of Dairy Science 93, 5188–5199.CrossRefGoogle ScholarPubMed
Demirel, I, Persson, A, Brauner, A, Särndahl, E, Kruse, R and Persson, K (2018) Activation of the NLRP3 inflammasome pathway by uropathogenic Escherichia coli is virulence factor-dependent and influences colonization of bladder epithelial cells. Frontiers in Cellular and Infection Microbiology 8, 81 doi: 10.3389/fcimb.2018.00081.CrossRefGoogle ScholarPubMed
Franchi, L, Muñoz-Planillo, R and Núñez, G (2012) Sensing and reacting to microbes through the inflammasomes. Nature Immunology 13, 325–332.CrossRefGoogle ScholarPubMed
Fu, Y, Gao, R, Cao, Y, Guo, M, Wei, Z, Zhou, E, Li, Y, Yao, M, Yang, Z and Zhang, N (2014) Curcumin attenuates inflammatory responses by suppressing TLR4-mediated NF-κB signaling pathway in lipopolysaccharide-induced mastitis in mice. International Immunopharmacology 20, 54–58.CrossRefGoogle ScholarPubMed
Gomes, F and Henriques, M (2016) Control of bovine mastitis: old and recent therapeutic approaches. Current Microbiology 72, 377–382.CrossRefGoogle ScholarPubMed
Griesbeckzilch, B, Meyer, HH, Kühn, CH, Schwerin, M and Wellnitz, O (2008) Staphylococcus aureus and Escherichia coli cause deviating expression profiles of cytokines and lactoferrin messenger ribonucleic acid in mammary epithelial cells. Journal of Dairy Science 91, 2215–2224.CrossRefGoogle Scholar
Guo, H, Callaway, JB and Ting, JP (2015) Inflammasomes: mechanism of action, role in disease, and therapeutics. Nature Medicine 21, 677–687.CrossRefGoogle Scholar
Gussmann, M, Steeneveld, W, Kirkeby, C, Hogeveen, H, Farre, M and Halasa, T (2019) Economic and epidemiological impact of different intervention strategies for subclinical and clinical mastitis. Preventive Veterinary Medicine 166, 78–85.CrossRefGoogle ScholarPubMed
Gutsmann, T, Müller, M, Carroll, SF, Mackenzie, RC, Wiese, A and Seydel, U (2001) Dual role of lipopolysaccharide (LPS)-binding protein in neutralization of LPS and enhancement of LPS-induced activation of mononuclear cells. Infection and Immunity 69, 6942.CrossRefGoogle ScholarPubMed
Günther, J, Petzl, W, Zerbe, H, Schuberth, HJ and Seyfert, HM (2016) TLR ligands, but not modulators of histone modifiers, can induce the complex immune response pattern of endotoxin tolerance in mammary epithelial cells. Innate Immunity 23, 155–164.CrossRefGoogle Scholar
Hertl, JA, Schukken, YH, Welcome, FL, Tauer, LW and Gröhn, YT (2014) Pathogen-specific effects on milk yield in repeated clinical mastitis episodes in Holstein dairy cows. Journal of Dairy Science 97, 1465–1480.CrossRefGoogle ScholarPubMed
Ibeaghaawemu, EM, Lee, JW, Ibeagha, AE, Bannerman, DD, Paape, MJ and Zhao, X (2008) Bacterial lipopolysaccharide induces increased expression of toll-like receptor (TLR) 4 and downstream TLR signaling molecules in bovine mammary epithelial cells. Veterinary Research 39, 11.CrossRefGoogle Scholar
Im, J, Lee, T, Jeon, JH, Baik, JE, Kim, KW, Kang, SS, Yun, CH, Kim, H and Han, SH (2014) Gene expression profiling of bovine mammary gland epithelial cells stimulated with lipoteichoic acid plus peptidoglycan from Staphylococcus aureus. International Immunopharmacology 21, 231–240.CrossRefGoogle ScholarPubMed
Jiang, L, Sørensen, P, Røntved, C, Vels, L and Ingvartsen, KL (2008) Gene expression profiling of liver from dairy cows treated intra-mammary with lipopolysaccharide. BMC Genomics 9, 443–443.CrossRefGoogle ScholarPubMed
Jo, EK, Kim, JK, Shin, DM and Sasakawa, C (2016) Molecular mechanisms regulating NLRP3 inflammasome activation. Cellular and Molecular Immunology 13, 148–159.CrossRefGoogle ScholarPubMed
Kang, S, Lee, JS, Lee, HC, Petriello, MC, Kim, BY, Do, JT, Lim, DS, Lee, HG and Han, SG (2016) Phytoncide extracted from pinecone decreases LPS induced inflammatory responses in bovine mammary epithelial cells. Journal of Microbiology and Biotechnology 26, 579–587.CrossRefGoogle ScholarPubMed
Lemarchand, E, Barrington, J, Chenery, A, Haley, M, Coutts, G, Allen, JE, Allan, SM and Brough, D (2019) Extent of ischemic brain injury after thrombotic stroke is independent of the NLRP3 (NACHT, LRR and PYD domains-containing protein 3) inflammasome. Stroke 50, 1232–1239.CrossRefGoogle ScholarPubMed
Li, CM, Wang, XL, Kuang, MQ, Li, L, Wang, YR, Yang, FX and Wang, GL (2019) UFL1 modulates NLRP3 inflammasome activation and protects against pyroptosis in LPS-stimulated bovine mammary epithelial cells. Molecular Immunology 112, 1–9.CrossRefGoogle ScholarPubMed
Liu, YG, Chen, JK, Zhang, ZT, Ma, XJ, Chen, YC, Du, XM, Liu, H, Zong, Y and Lu, GC (2017) NLRP3 inflammasome activation mediates radiation-induced pyroptosis in bone marrow-derived macrophages. Cell Death & Disease 8, e2579.CrossRefGoogle ScholarPubMed
Mariathasan, S and Monack, DM (2007) Inflammasome adaptors and sensors: intracellular regulators of infection and inflammation. Nature Reviews Immunology 7, 31–40.CrossRefGoogle ScholarPubMed
Ozaki, E, Campbell, M and Doyle, SL (2015) Targeting the NLRP3 inflammasome in chronic inflammatory diseases: current perspectives. Journal of Inflammation Research 8, 15–27.Google ScholarPubMed
Philpott, DJ, Girardin, SE and Sansonetti, PJ (2001) Innate immune responses of epithelial cells following infection with bacterial pathogens. Current Opinion in Immunology 13, 410–416.CrossRefGoogle ScholarPubMed
Porcherie, A, Cunha, P, Trotereau, A, Roussel, P, Gilbert, FB, Rainard, P, Gilbert, FB, Rainard, P and Germon, P (2012) Repertoire of Escherichia coli agonists sensed by innate immunity receptors of the bovine udder and mammary epithelial cells. Veterinary Research 43, 14.CrossRefGoogle ScholarPubMed
Qu, S, Wang, W, Li, D, Li, S, Zhang, L, Fu, Y and Zhang, N (2017) Mangiferin inhibits mastitis induced by LPS via suppressing NF-κB and NLRP3 signaling pathways. International Immunopharmacology 43, 85–90.CrossRefGoogle ScholarPubMed
Seegers, H, Fourichon, C and Beaudeau, F (2003) Production effects related to mastitis and mastitis economics in dairy cattle herds. Veterinary Research 34, 475–491.CrossRefGoogle ScholarPubMed
Shaftel, SS, Carlson, TJ, Olschowka, JA, Kyrkanides, S, Matousek, SB and O'Banion, MK (2007) Chronic interleukin-1beta expression in mouse brain leads to leukocyte infiltration and neutrophil-independent blood brain barrier permeability without overt neurodegeneration. Journal of Neuroscience 27, 9301–9309.CrossRefGoogle ScholarPubMed
Strandberg, Y, Gray, C, Vuocolo, T, Donaldson, L, Broadway, M and Tellam, R (2005) Lipopolysaccharide and lipoteichoic acid induce different innate immune responses in bovine mammary epithelial cells. Cytokine 31, 72–86.CrossRefGoogle ScholarPubMed
Sun, Y, Li, L, Wu, J, Yu, P, Li, C, Tang, J, Li, XJ, Huang, S and Wang, GL (2015) Bovine recombinant lipopolysaccharide binding protein (BRLBP) regulated apoptosis and inflammation response in lipopolysaccharide-challenged bovine mammary epithelial cells (BMEC). Molecular Immunology 65, 205–214.CrossRefGoogle Scholar
Wang, X, Feng, S, Ding, N, He, Y, Li, C, Li, M, Ding, X, Ding, H, Li, J, Wu, J and Li, Y (2018) Anti-inflammatory effects of berberine hydrochloride in an LPS-induced murine model of mastitis. Evidence-Based Complementary and Alternative Medicine, 2018, 1–9.Google Scholar
Wang, YJ, Gong, GQ, Chen, S, Xiong, LY, Zhou, XX, Huang, X and Kong, WJ (2014) NLRP3 inflammasome sequential changes in Staphylococcus aureus-induced mouse model of acute rhinosinusitis. International Journal of Molecular Sciences 15, 15806–15820.CrossRefGoogle ScholarPubMed
Wenting, D, Quanjuan, W, Fengqi, Z, Jianxin, L and Hongyun, L (2018) Understanding the regulatory mechanisms of milk production using integrative transcriptomic and proteomic analyses: improving inefficient utilization of crop by-products as forage in dairy industry. BMC Genomics 19, 403.Google Scholar
Yang, W, Zerbe, H, Petzl, W, Brunner, RM, Günther, J, Draing, C, von Aulock, S, Schuberth, HJ and Seyfert, HM (2008) Bovine TLR2 and TLR4 properly transduce signals from Staphylococcus aureus and E. coli, but S. aureus fails to both activate NF-kappaB in mammary epithelial cells and to quickly induce TNFalpha and interleukin-8 (cxcl8) expression in the udder. Molecular Immunology 45, 1385–1397.CrossRefGoogle Scholar
Yang, B, Zhou, Z, Li, X and Niu, J (2016) The effect of lysophosphatidic acid on toll-like receptor 4 expression and the nuclear factor-κB signaling pathway in THP-1 cells. Molecular and Cellular Biochemistry 422, 1–9.CrossRefGoogle ScholarPubMed
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