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 |
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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. 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><subject>Cattle</subject><subject>Cell growth</subject><subject>Cell proliferation</subject><subject>Change detection</subject><subject>Cytokines</subject><subject>Dairy cattle</subject><subject>Dairy products</subject><subject>Enzyme-linked immunosorbent assay</subject><subject>Epithelial cells</subject><subject>Flow cytometry</subject><subject>Food quality</subject><subject>Food safety</subject><subject>FSCN1</subject><subject>Gene expression</subject><subject>Infections</subject><subject>Inflammation</subject><subject>Interferon</subject><subject>Lymphocytes</subject><subject>Lymphocytes T</subject><subject>Mammary gland</subject><subject>Mammary glands</subject><subject>Mastitis</subject><subject>Microorganisms</subject><subject>miRNA</subject><subject>miR‐145</subject><subject>Ribonucleic acid</subject><subject>RNA</subject><subject>Staphylococcus aureus</subject><subject>Staphylococcus infections</subject><subject>Tumor necrosis factor</subject><subject>Western blotting</subject><issn>0936-6768</issn><issn>1439-0531</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2019</creationdate><recordtype>article</recordtype><recordid>eNp10ctKAzEUBuAgCtbqwjcIuNHFtMkkM5MsS7Uq1Aqtroc0k9bUudRcLLPzEXxGn8TUcSWYzSGH7xxCfgDOMRrgcIamEANMKGEHoIcp4RFKCD4EPcRJGqVZyo7BibUbhHDCsqwHdg9ammY-G319fGKaQKPWvhROWairytcKytY1r7oOjXctoBNmrZyu13CyGM8w1DVcOLF9actGNlJ6C4U3ytuwTdeFl6qAlbBOO233thDatFA2O3sKjlaitOrst_bB8-TmaXwXTR9v78ejaSQJQSxa8jgrBMeSJRmnNGMkVZKmSPIYC1QQRXCiRLhQJpRIqVwWGWIUx5zJVYI56YPLbu_WNG9eWZdX2kpVlqJWjbd5HCOeojCfBnrxh24ab-rwuqAoJowmeK-uOhW-zVqjVvnW6EqYNsco30eQhwjynwiCHXZ2p0vV_g_z-fWom_gGYkyJIQ</recordid><startdate>201906</startdate><enddate>201906</enddate><creator>Chen, Zhi</creator><creator>Xu, Xin</creator><creator>Tan, Taoling</creator><creator>Chen, Daijie</creator><creator>Liang, Huanjie</creator><creator>Sun, Kaidi</creator><creator>Li, Mingxun</creator><creator>Zhang, Huimin</creator><creator>Mao, Yongjiang</creator><creator>Yang, Zhangping</creator><general>Blackwell Publishing Ltd</general><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope><orcidid>https://orcid.org/0000-0003-3924-1891</orcidid></search><sort><creationdate>201906</creationdate><title>MicroRNA‐145 regulates immune cytokines via targeting FSCN1 in Staphylococcus aureus‐induced mastitis in dairy cows</title><author>Chen, Zhi ; Xu, Xin ; Tan, Taoling ; Chen, Daijie ; Liang, Huanjie ; Sun, Kaidi ; Li, Mingxun ; Zhang, Huimin ; Mao, Yongjiang ; Yang, Zhangping</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3308-b927da91c8579447836ec460c921a0d3e315ea92148aea64cbd70841298cf5193</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2019</creationdate><topic>Cattle</topic><topic>Cell growth</topic><topic>Cell proliferation</topic><topic>Change detection</topic><topic>Cytokines</topic><topic>Dairy cattle</topic><topic>Dairy products</topic><topic>Enzyme-linked immunosorbent assay</topic><topic>Epithelial cells</topic><topic>Flow cytometry</topic><topic>Food quality</topic><topic>Food safety</topic><topic>FSCN1</topic><topic>Gene expression</topic><topic>Infections</topic><topic>Inflammation</topic><topic>Interferon</topic><topic>Lymphocytes</topic><topic>Lymphocytes T</topic><topic>Mammary gland</topic><topic>Mammary glands</topic><topic>Mastitis</topic><topic>Microorganisms</topic><topic>miRNA</topic><topic>miR‐145</topic><topic>Ribonucleic acid</topic><topic>RNA</topic><topic>Staphylococcus aureus</topic><topic>Staphylococcus infections</topic><topic>Tumor necrosis factor</topic><topic>Western blotting</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><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><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Reproduction in domestic animals</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chen, Zhi</au><au>Xu, Xin</au><au>Tan, Taoling</au><au>Chen, Daijie</au><au>Liang, Huanjie</au><au>Sun, Kaidi</au><au>Li, Mingxun</au><au>Zhang, Huimin</au><au>Mao, Yongjiang</au><au>Yang, Zhangping</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>MicroRNA‐145 regulates immune cytokines via targeting FSCN1 in Staphylococcus aureus‐induced mastitis in dairy cows</atitle><jtitle>Reproduction in domestic animals</jtitle><date>2019-06</date><risdate>2019</risdate><volume>54</volume><issue>6</issue><spage>882</spage><epage>891</epage><pages>882-891</pages><issn>0936-6768</issn><eissn>1439-0531</eissn><abstract>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.</abstract><cop>Oxford</cop><pub>Blackwell Publishing Ltd</pub><doi>10.1111/rda.13438</doi><tpages>10</tpages><orcidid>https://orcid.org/0000-0003-3924-1891</orcidid></addata></record> |
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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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