MicroRNA‐145 regulates immune cytokines via targeting FSCN1 in Staphylococcus aureus‐induced mastitis in dairy cows

Dairy cow mastitis is a detrimental factor in milk quality and food safety. Mastitis generally refers to inflammation caused by infection by pathogenic microorganisms. Our studies in recent years have revealed the role of miRNA regulation in Staphylococcus aureus‐induced mastitis. In the present stu...

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Veröffentlicht in:Reproduction in domestic animals 2019-06, Vol.54 (6), p.882-891
Hauptverfasser: Chen, Zhi, Xu, Xin, Tan, Taoling, Chen, Daijie, Liang, Huanjie, Sun, Kaidi, Li, Mingxun, Zhang, Huimin, Mao, Yongjiang, Yang, Zhangping
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container_end_page 891
container_issue 6
container_start_page 882
container_title Reproduction in domestic animals
container_volume 54
creator Chen, Zhi
Xu, Xin
Tan, Taoling
Chen, Daijie
Liang, Huanjie
Sun, Kaidi
Li, Mingxun
Zhang, Huimin
Mao, Yongjiang
Yang, Zhangping
description Dairy cow mastitis is a detrimental factor in milk quality and food safety. Mastitis generally refers to inflammation caused by infection by pathogenic microorganisms. Our studies in recent years have revealed the role of miRNA regulation in Staphylococcus aureus‐induced mastitis. In the present study, we overexpressed and suppressed miR‐145 to investigate the function of miR‐145 in Mac‐T cells. Flow cytometry, ELISA and EdU staining were used to detect changes in the secretion of several Mac‐T cytokines and in cell proliferation. We found that overexpression of miR‐145 in Mac‐T cells significantly reduced the secretion of IL‐12 and TNF‐α, but increased the secretion of IFN‐γ; the proliferation of bovine mammary epithelial cells was also inhibited. Using quantitative real‐time PCR (qRT‐PCR), Western blotting and luciferase multiplex verification techniques, we found that miR‐145 targeted and regulated FSCN1. Knock‐down of FSCN1 significantly increased the secretion of IL‐12, while the secretion of TNF‐α was significantly downregulated in Mac‐T cells. Upon S. aureus infection of mammary gland tissue, the body initiated inflammatory responses; Bta‐miR‐145 expression was downregulated, which reduced the inhibitory effect on the FSCN1 gene; and upregulation of FSCN1 expression promoted mammary epithelial cell proliferation to allow the recovery of damaged tissue. The results of the present study will aid in understanding the immune mechanism opposing S. aureus infection in dairy cows and will provide a laboratory research basis for the prevention and treatment of mastitis.
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Mastitis generally refers to inflammation caused by infection by pathogenic microorganisms. Our studies in recent years have revealed the role of miRNA regulation in Staphylococcus aureus‐induced mastitis. In the present study, we overexpressed and suppressed miR‐145 to investigate the function of miR‐145 in Mac‐T cells. Flow cytometry, ELISA and EdU staining were used to detect changes in the secretion of several Mac‐T cytokines and in cell proliferation. We found that overexpression of miR‐145 in Mac‐T cells significantly reduced the secretion of IL‐12 and TNF‐α, but increased the secretion of IFN‐γ; the proliferation of bovine mammary epithelial cells was also inhibited. Using quantitative real‐time PCR (qRT‐PCR), Western blotting and luciferase multiplex verification techniques, we found that miR‐145 targeted and regulated FSCN1. Knock‐down of FSCN1 significantly increased the secretion of IL‐12, while the secretion of TNF‐α was significantly downregulated in Mac‐T cells. Upon S. aureus infection of mammary gland tissue, the body initiated inflammatory responses; Bta‐miR‐145 expression was downregulated, which reduced the inhibitory effect on the FSCN1 gene; and upregulation of FSCN1 expression promoted mammary epithelial cell proliferation to allow the recovery of damaged tissue. The results of the present study will aid in understanding the immune mechanism opposing S. aureus infection in dairy cows and will provide a laboratory research basis for the prevention and treatment of mastitis.</description><identifier>ISSN: 0936-6768</identifier><identifier>EISSN: 1439-0531</identifier><identifier>DOI: 10.1111/rda.13438</identifier><language>eng</language><publisher>Oxford: Blackwell Publishing Ltd</publisher><subject>Cattle ; Cell growth ; Cell proliferation ; Change detection ; Cytokines ; Dairy cattle ; Dairy products ; Enzyme-linked immunosorbent assay ; Epithelial cells ; Flow cytometry ; Food quality ; Food safety ; FSCN1 ; Gene expression ; Infections ; Inflammation ; Interferon ; Lymphocytes ; Lymphocytes T ; Mammary gland ; Mammary glands ; Mastitis ; Microorganisms ; miRNA ; miR‐145 ; Ribonucleic acid ; RNA ; Staphylococcus aureus ; Staphylococcus infections ; Tumor necrosis factor ; Western blotting</subject><ispartof>Reproduction in domestic animals, 2019-06, Vol.54 (6), p.882-891</ispartof><rights>2019 Blackwell Verlag GmbH</rights><rights>Copyright © 2019 Blackwell Verlag GmbH</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3308-b927da91c8579447836ec460c921a0d3e315ea92148aea64cbd70841298cf5193</citedby><cites>FETCH-LOGICAL-c3308-b927da91c8579447836ec460c921a0d3e315ea92148aea64cbd70841298cf5193</cites><orcidid>0000-0003-3924-1891</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Frda.13438$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Frda.13438$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27903,27904,45553,45554</link.rule.ids></links><search><creatorcontrib>Chen, Zhi</creatorcontrib><creatorcontrib>Xu, Xin</creatorcontrib><creatorcontrib>Tan, Taoling</creatorcontrib><creatorcontrib>Chen, Daijie</creatorcontrib><creatorcontrib>Liang, Huanjie</creatorcontrib><creatorcontrib>Sun, Kaidi</creatorcontrib><creatorcontrib>Li, Mingxun</creatorcontrib><creatorcontrib>Zhang, Huimin</creatorcontrib><creatorcontrib>Mao, Yongjiang</creatorcontrib><creatorcontrib>Yang, Zhangping</creatorcontrib><title>MicroRNA‐145 regulates immune cytokines via targeting FSCN1 in Staphylococcus aureus‐induced mastitis in dairy cows</title><title>Reproduction in domestic animals</title><description>Dairy cow mastitis is a detrimental factor in milk quality and food safety. 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Upon S. aureus infection of mammary gland tissue, the body initiated inflammatory responses; Bta‐miR‐145 expression was downregulated, which reduced the inhibitory effect on the FSCN1 gene; and upregulation of FSCN1 expression promoted mammary epithelial cell proliferation to allow the recovery of damaged tissue. 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source Wiley Online Library - AutoHoldings Journals
subjects Cattle
Cell growth
Cell proliferation
Change detection
Cytokines
Dairy cattle
Dairy products
Enzyme-linked immunosorbent assay
Epithelial cells
Flow cytometry
Food quality
Food safety
FSCN1
Gene expression
Infections
Inflammation
Interferon
Lymphocytes
Lymphocytes T
Mammary gland
Mammary glands
Mastitis
Microorganisms
miRNA
miR‐145
Ribonucleic acid
RNA
Staphylococcus aureus
Staphylococcus infections
Tumor necrosis factor
Western blotting
title MicroRNA‐145 regulates immune cytokines via targeting FSCN1 in Staphylococcus aureus‐induced mastitis in dairy cows
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