Proteomic analysis of bovine mammary epithelial cells after in vitro incubation withS. agalactiae: potential biomarkers

文献类型： 外文期刊

第一作者： Tong, Jinjin

作者： Tong, Jinjin;Sun, Mingwei;Zhang, Hua;Yang, Delian;Zhang, Yonghong;Jiang, Linshu;Xiong, Benhai

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关键词： proteomic analysis; bovine mammary epithelial cells; Streptococcus agalactiae; potential biomarkers

期刊名称：VETERINARY RESEARCH （ 影响因子：3.683； 五年影响因子：4.106 ）

ISSN： 0928-4249

年卷期： 2020 年 51 卷 1 期

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收录情况： SCI

摘要： Streptococcus agalactiaeis one of the causative agents of subclinical mastitis, a common disease of dairy cows that causes great economic losses in the industry worldwide. It is thought that pathology is mainly due to inflammatory damage of bovine mammary epithelial cells (bMECs); however, the mechanism by whichS. agalactiaedamages the bMECs is not clear. The aim of this study was to evaluate the inflammatory effects ofS. agalactiaeon bMECs and the resulting changes in protein profiles. The bMECs were incubated withS. agalactiaefor different times and assayed for cell viability by MTT assay, apoptosis by annexin V and propidium iodide dual staining, and morphological and ultrastructural changes by scanning and transmission electron microscopy. Quantitative real-time PCR was used to determine the effect ofS. agalactiaeon expression of mRNA of inflammatory factors in bMECs and protein levels were quantitated by liquid chromatography/mass spectrometry. Exposure toS. agalactiaesignificantly decreased the cell viability and triggered apoptosis, as well as up-regulating TNF-alpha, IL-1 beta and IL-6 mRNA, and inhibiting IL-8 expression.S. agalactiaealso induced morphological and ultrastructural changes. Furthermore, we identified 325 up-regulated and 704 down-regulated proteins in the treated vs control group. All significant differentially expressed proteins (DSEPs) were classified into three major areas by function: biological processes, cellular components and molecular functions. These differentially expressed proteins included enzymes and proteins associated with various metabolic processes and cellular immunity. Pathway enrichment analysis showed that eight down-regulated signaling pathways were significantly enriched. Exposure to even subclinical levels ofS. agalactiaecan lead to inflammation and bMEC damage. Our data suggest some possible molecular mechanisms for the harmful effects of subclinical mastitis in dairy cows.

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