MicroRNA-145 regulates immune cytokines via targeting FSCN1 in Staphylococcus aureus-induced mastitis in dairy cows
MicroRNA-145 regulates immune cytokines via targeting FSCN1 in Staphylococcus aureus-induced mastitis in dairy cows
复制标题
MicroRNA-145通过靶向FSCN1在金黄色葡萄球菌引起的奶牛乳腺炎中调节免疫细胞因子
DOI：
10.1111/rda.13438
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发表时间：
2019-06-01
影响因子：
1.7
通讯作者：
Yang, Zhangping
中科院分区：
文献类型：
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作者：
Chen, Zhi;Xu, Xin;Yang, Zhangping
奶牛乳房炎是影响牛奶质量和食品安全的重要因素。乳腺炎通常是指由病原微生物感染引起的炎症。我们近年来的研究揭示了miRNA调控在金黄色葡萄球菌诱导的乳腺炎中的作用。在本研究中，我们过表达和抑制miR-145，以研究miR-145在Mac-T细胞中的功能。流式细胞术、ELISA和EdU染色用于检测几种Mac-T细胞因子分泌和细胞增殖的变化。我们发现，在Mac-T细胞中过表达miR-145显著降低了IL-12和TNF-α的分泌，但增加了IFN-γ的分泌;牛乳腺上皮细胞的增殖也受到抑制。利用实时定量PCR（qRT-PCR）、Western blotting和荧光素酶多重验证技术，我们发现miR-145靶向并调节FSCN 1。敲低FSCN 1显著增加IL-12的分泌，而Mac-T细胞中TNF-α的分泌显著下调。在S.金黄色葡萄球菌感染乳腺组织后，机体启动炎症反应; Bta-miR-145表达下调，降低了对FSCN 1基因的抑制作用; FSCN 1表达上调促进乳腺上皮细胞增殖，使受损组织得以恢复。本研究结果将有助于了解抗S.奶牛金黄色葡萄球菌感染的研究，将为乳腺炎的防治提供实验室研究依据。
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-alpha, but increased the secretion of IFN-gamma; 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-alpha 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.