Proteomics study on the protective mechanism of soybean isoflavone against inflammation injury of bovine mammary epithel
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ORIGINAL PAPER
Proteomics study on the protective mechanism of soybean isoflavone against inflammation injury of bovine mammary epithelial cells induced by Streptococcus agalactiae Hui Niu 1
&
Hua Zhang 1
&
Fuxin Wu 1
&
Benhai Xiong 2
&
Jinjin Tong 1
&
Linshu Jiang 1
Received: 19 June 2020 / Revised: 21 August 2020 / Accepted: 25 August 2020 # The Author(s) 2020
Abstract This study aimed to verify the anti-inflammatory effect of soybean isoflavones (SI) on the inflammatory response induced by Streptococcus agalactiae (S. agalactiae) of bovine mammary epithelial cells (bMECs) and to elucidate its possible mechanism. BMECs were pretreated with SI of different concentrations (20, 40, 60, 80, 100 μg/mL) for 0.5, 3, 6, 9, 12, 15, 18, 24 h. And then, S. agalactiae was used to infect bMECs for 6 h (MOI = 50:1) to establish the inflammation model. Cell viability, growth curves of S. agalactiae, cytotoxicity, and S. agalactiae invasion rate were determined. A proteomics technique was used to further detect differential proteins and enrichment pathways. SI (40 μg/mL) improved the viability of bMECs at 12 h (p < 0.05) and 60 and 80 μg/mL of SI greater (p < 0.01). Moreover, 60 μg/mL of SI protects cells from bacterial damage (p < 0.05). SI could inhibit S. agalactiae growth and internalization into bMECs in a time- and dose-dependent manner. In addition, proteomics results showed that 133 proteins were up-regulated and 89 proteins were down-regulated significantly. The differentially significantly expressed proteins (DSEPs) were mainly related to cell proliferation, differentiation, apoptosis, and migration. GO annotation showed that 222 DSEPs were divided into 23 biological processes (BP) terms, 14 cell components (CC) terms, and 12 molecular functions (MF) terms. DSEPs were significantly enriched in 10 pathways, of which the immune pathway was the main enrichment pathway. Keywords Soybean isoflavone . Dairy mammary epithelial cells . Streptococcus agalactiae . Protection mechanism . Proteomics
Introduction S. agalactiae is one of the common pathogens causing subclinical mastitis in dairy cows (Santos et al. 2011). Mammary epithelial cells, as the first line of defense against mastitis, play an important role in the process of resisting the invasion of pathogenic microorganisms (Bougarn et al. 2011). After dairy cows are infected with S. agalactiae, bacteria in the infected Hui Niu and Hua Zhang contributed equally to this work. * Jinjin Tong [email protected] * Linshu Jiang [email protected] 1
Department of Animal Science, Animal Science and Technology College, Beijing University of Agriculture, Beijing 102206, China
2
State Key Laboratory of Animal Nutrition, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China
mammary gland gradually increase. In the case of continuous infection, somatic cell count in milk is increasing, and mammary gland tissue is further damaged. The acini in the gland finally lose their complete structure, and the blood milk barrier is
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