Proteomics and Metabolomics Study on the Responses of Sertoli Cells Infected With Brucella and Its bvfA-Deletion Strains

文献类型： 外文期刊

第一作者： Jia, Fang

作者： Jia, Fang;Yang, Jiangliu;Wang, Yujiong;Zhou, Xuezhang;Jia, Fang;Liu, Jun

作者机构：

关键词： Brucella abortus S19; Brucella virulence factor A (BvfA); label-free; LC-MS; sertoli cells (TM4)

期刊名称：PROTEOMICS CLINICAL APPLICATIONS （ 影响因子：2.5； 五年影响因子：2.4 ）

ISSN： 1862-8346

年卷期： 2025 年 19 卷 1 期

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收录情况： SCI

摘要： ObjectiveTo investigate the potential effects of BvfA in reproductive system damage caused by Brucella.MethodsBrucella intracellular multiplication ability was determined by a gentamicin protection assay; the LDH method was used to determine the lethal effect of Brucella on TM4 cells. Afterward, Label-free proteomics and LC-MS/MS metabolomics assays were combined to reveal differential abundant proteins and metabolites of TM4 cells infected with bvfA-deletion strains and parental strains. Finally, PRM mass spectrometry and western blot analysis were carried out to confirm differential expression of proteins.ResultsThis report demonstrated that bvfA-deletion strains failed to invade TM4 cells and reconstitution of invasion when a strain with gene bvfA was reintroduced to the deletion strain in 3 h. The bvfA-deletion exhibited weakened intracellular multiplication compared with parental strains in TM4 cells in 12 h; however, the death rate of TM4 cells infected with bvfA-deletion strains was higher than that of TM4 cells infected with parental strains. Combined proteomics and metabolomics analyses revealed that the differential abundant proteins and metabolites in TM4 cells infected with bvfA-deletion and parental strains mainly involved the mineral absorption-related pathway, NADH:ubiquinone oxidoreductase subunit-related mitochondrial respiratory signaling pathway, and sphingolipid signaling pathway of TM4 cells. These three signaling pathways were involved in expression changes of TRPM6/7, STEAP1, Gnaq, Trp53, Pbk, Tns2, Akt2, and the NADH:ubiquinone oxidoreductase subunit, as well as content changes of l-Valine, l-Isoleucine, l-Methionine, PC, PE DG, and SM metabolites.SignificanceThese results indicated that BvfA of Brucella abortus S19 affected the above proteins and metabolites in TM4 cells.

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