基于多组学解析布鲁氏菌BvfA对睾丸支持细胞线粒体NADH-泛醌氧化还原酶复合体的影响  

Multi-omics Analysis of the Effect of Brucella Virulence Factor A on the NADH-ubiquinone Oxidoreductase ComplexⅠof Mitochondria in Sertoli Cells

作　　者：宋青山[1] 杨江流 刘军[4] 贾芳 SONG Qingshan;YANG Jiangliu;LIU Jun;JIA Fang(Hetao College,Bayannur 015000,China;Inner Mongolia Key Laboratory of Molecular Biology,School of Basic Medical Sciences,Inner Mongolia Medical University,Hohhot 010110,China;Key Laboratory of the Ministry of Education for the Conservation and Utilization of Special Biological Resources of Western China,School of Life Sciences,Ningxia University,Yinchuan 750021,China;Changchun Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Changchun 130122,China)

机构地区：[1]河套学院,巴彦淖尔015000 [2]内蒙古医科大学基础医学院,内蒙古自治区分子生物学重点实验室,呼和浩特010110 [3]宁夏大学生命科学学院,西部特色生物资源保护与利用教育部重点实验室,银川750021 [4]中国农业科学院长春兽医研究所,长春130122

出　　处：《中国畜牧兽医》2024年第4期1400-1409,共10页China Animal Husbandry & Veterinary Medicine

基　　金：内蒙古自治区高等学校科学技术研究项目(NJZZ21059、NJZZ23003)。

摘　　要：【目的】明确布鲁氏菌BvfA对宿主睾丸支持细胞(TM4细胞)的损伤作用,以揭示布鲁氏菌对宿主的致病机制。【方法】利用RNA-Seq双端测序、Label-free蛋白组学技术和LC-MS非靶代谢组学技术分析感染布鲁氏菌S19△bvfA与S19菌株的TM4细胞的组学差异,采用KEGG数据库对差异表达蛋白及差异代谢物通路进行分析,并利用平行反应监测(PRM)对蛋白表达量进行验证。【结果】感染S19△bvfA和S19菌株的TM4细胞差异表达基因共400个,差异表达蛋白共422个,差异代谢产物共271种。多组学联合分析发现,S19△bvfA和S19菌株感染TM4细胞差异表达基因、差异表达蛋白和差异代谢产物共同富集的KEGG通路为内源性大麻素信号通路,在该通路中发现S19△bvfA菌株感染TM4细胞的线粒体NADH-泛醌氧化还原酶复合体(ComplexⅠ)中有20个蛋白表达量下调。PRM验证结果显示,ComplexⅠ中Ndufb11、Ndufa9、Ndufv1、Ndufv2、Ndufs8和Ndufs2蛋白的表达趋势与Label-free蛋白组学技术测定结果一致。【结论】BvfA在布鲁氏菌感染TM4细胞时能引起转录、蛋白质和代谢水平的变化,布鲁氏菌BvfA使睾丸支持细胞线粒体NADH-泛醌氧化还原酶复合体蛋白表达下调。【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 ComplexⅠof TM4 cells infected with S19△bvfA strain.PRM result showed that the expression trends of Ndufb11,Ndufa9,Ndufv1,Ndufv2,Ndufs8 and Ndufs2 in ComplexⅠwere 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.

关 键 词：布鲁氏菌 多组学 BvfA 睾丸支持细胞 

分 类 号：S852.61[农业科学—基础兽医学]