Multi-omics Analysis of the Effect of Brucella Virulence Factor A on the NADH-ubiquinone Oxidoreductase Complex Ⅰ of Mitochondria in Sertoli Cells
[Objective]This study was aimed to investigate the damage effect of Brucella virulence factor A(BvfA)in Sertoli cells(TM4 cells),so as to reveal the pathogenic mechanism of Brucella on host.[Method]RNA-Seq double-end sequencing,Label-free proteomics and LC-MS non-target metabolomics were used to analyze the omics difference of TM4 cells infected with Brucella S19 △bvfA and S19 strains.The differentially expressed proteins and differential metabolite pathways were analyzed using KEGG database,and the protein expression were validated by parallel reaction monitoring(PRM).[Result]There were 400 differentially expressed genes,422 differentially expressed proteins and 271 differential metabolites in TM4 cells infected with S19△bvfA and S19 strains.Multi-omics analysis showed that the common KEGG pathway of differentially expressed genes,proteins and metabolites in TM4 cells infected with S19 △bvfA and S19 strains was retrograde endocannabinoid signaling pathway.In this pathway,20 proteins expression was down-regulated in mitochondrial NADH-ubiquinone redoxase Complex Ⅰ of TM4 cells infected with S1 9△bvfA strain.PRM result showed that the expression trends of Ndufb11,Ndufa9,Ndufv1,Ndufv2,Ndufs8 and Ndufs2 in Complex Ⅰ were consistent with the results of Label-free proteomics.[Conclusion]BvfA induced variations of transcriptional,protein and metabolic levels in Brucella infected TM4 cells,and BvfA down-regulated the expression of NADH-ubiquinone oxidoreductase complex proteins in mitochondria of Sertoli cells.
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