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

doi:10.16431/j.cnki.1671-7236.2024.04.008

中国畜牧兽医 ›› 2024, Vol. 51 ›› Issue (4): 1400-1409.doi: 10.16431/j.cnki.1671-7236.2024.04.008

• 生物技术 • 上一篇

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

宋青山¹, 杨江流³, 刘军⁴, 贾芳²

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

收稿日期:2023-09-19 发布日期:2024-03-27
通讯作者: 刘军, 贾芳 E-mail:liubio@126.com;20220004@immu.edu.cn
作者简介:宋青山,E-mail:3036374116@qq.com。
基金资助:
内蒙古自治区高等学校科学技术研究项目(NJZZ21059、NJZZ23003)

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

SONG Qingshan¹, YANG Jiangliu³, LIU Jun⁴, JIA Fang²

1. Hetao College, Bayannur 015000, China;
2. Inner Mongolia Key Laboratory of Molecular Biology, School of Basic Medical Sciences, Inner Mongolia Medical University, Hohhot 010110, China;
3. 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;
4. Changchun Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Changchun 130122, China

Received:2023-09-19 Published:2024-03-27

摘要/Abstract

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(Complex Ⅰ)中有20个蛋白表达量下调。PRM验证结果显示,Complex Ⅰ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。

关键词: 布鲁氏菌; 多组学; BvfA; 睾丸支持细胞

Abstract: 【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 S19△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.

Key words: Brucella; multi-omics; BvfA; Sertoli cells

中图分类号:

S852.61⁺4

宋青山, 杨江流, 刘军, 贾芳. 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[J]. 中国畜牧兽医, 2024, 51(4): 1400-1409.

SONG Qingshan, YANG Jiangliu, LIU Jun, JIA Fang. Multi-omics Analysis of the Effect of Brucella Virulence Factor A on the NADH-ubiquinone Oxidoreductase Complex Ⅰ of Mitochondria in Sertoli Cells[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(4): 1400-1409.

参考文献

[1] YAN X,HU S,YANG Y,et al.Proteomics investigation of the time course responses of RAW264.7 macrophages to infections with the wild-type and twin-arginine translocation mutant strains of Brucella melitensis[J].Frontiers in Cellular and Infection Microbiology,2021,11(6):679571.
[2] YU J,LI S,WANG L,et al.Pathogenesis of Brucella epididymoorchitis-game of Brucella death[J].Critical Reviews in Microbiology,2022,48(1):96-120.
[3] NAZ H,KORKMAZ P,CEVIK F,et al.The clinical and laboratory characteristics,treatments,and outcomes of patients with Brucella epididymoorchitis (BEO) compared to those without BEO[J]. Turkish Journal of Medical Sciences,2016,46(5):1323-1328.
[4] MKSKYTTASEON S.Brucella epididymoorchitis initially mimicking choledocholithiasis[J].Infection & Immunity,2011,39(1):393-394.
[5] MATHUR S,BANAI M,COHEN D.Natural Brucella melitensis infection and Rev.1 vaccination induce specific Brucella O-polysaccharide antibodies involved in complement mediated Brucella cell killing[J].Vaccines(Basel),2022,10(2):317.
[6] LAVIGNE J P,PATEY G,SANGARI F J, et al.Identification of a new virulence factor,BvfA,in Brucella suis[J].Infection and Immunity,2005,73(9):5524-5529.
[7] 贾芳,王鑫,王玉炯,等.bvfA基因对布鲁氏菌S19株生物学特性及睾丸支持细胞影响的研究[J].中国预防兽医学报,2022,44(6):611-616. JIA F,WANG X,WANG Y J,et al.Influence of bvfA gene on the biological characteristics of B.abortus S19 strain and Sertoli cells[J].Chinese Journal of Preventive Veterinary Medicine,2022,44(6):611-616.(in Chinese)
[8] LIU X,NIE S,CHEN Y,et al.Effects of 4-nonylphenol isomers on cell receptors and mitogen-activated protein kinase pathway in mouse Sertoli TM4 cells[J].Toxicology,2014,326(9):1-8.
[9] ZHANG Q,XIE X,LIU Y,et al.Fructose as an additional co-metabolite promotes refractory dye degradation:Performance and mechanism[J].Bioresource Technology,2019,46(2):430-440.
[10] WANG X,SUN G,FENG T,et al.Sodium oligomannate therapeutically remodels gut microbiota and suppresses gut bacterial amino acids-shaped neuroinflammation to inhibit Alzheimer’s disease progression[J].Cell Research,2019,29(6):787-803.
[11] LIU H,LIU X,LI X,et al.The molecular regulatory mechanisms of the bacteria involved in serpentine weathering coupled with carbonation[J].Chemical Geology,2021,565:120069.
[12] LAI S,ZHOU H,XIONG W,et al.Changing epidemiology of human brucellosis,China,1955-2014[J].Eemerging Infectious Disease,2017,23(2):184-194.
[13] JIANG H,O’CALLAGHAN D,DING J B.Brucellosis in China:History,progress and challenge[J].Infectious Diseases of Poverty,2020,55(9):55.
[14] TA N,YU R,LIAN H,et al.Analysis of laboratory and serological test results in patients with acute brucellosis during follow-up[J].Journal of Clinical Laboratory Analysis,2022,36:e24205.
[15] BRIDGES H R,FEDOR J G,BLAZA J N,et al.Testicular infection in brucellosis:Report of 34 cases[J].Journal of Microbiology Immunology and Infection,2018,51:82-87.
[16] ZHANG Y,ZHANG Y,ZHONG C,et al.Cr(VI) induces premature senescence through ROS-mediated p53 pathway in L-02 hepatocytes[J].Scientific Reports,2016,6:34578.
[17] BRIDGES H R,FEDOR J G,BLAZA J N,et al.Structure of inhibitor-bound mammalian complex Ⅰ[J].Nature Communications,2020,11:5261.
[18] BAKHTINA A A,PHARAOH G A,CAMPBELL M D,et al.Skeletal muscle mitochondrial interactome remodeling is linked to functional decline in aged female mice[J].Nature Aging,2023,3(3):313-326.
[19] NUBER F,SCHIMPF J,DI RAGO J P,et al.Biochemical consequences of two clinically relevant ND-gene mutations in Escherichia coli respiratory complex Ⅰ[J].Scientific Reports,2021,11:12641
[20] VERBEKE J,FAYT Y,MARTIN L,et al.Host cell egress of Brucella abortus requires BNIP3L-mediated mitophagy[J].Journal of the European Molecular Biology Society,2023,42:e112817.
[21] LOBET E,WILLEMART K,NINANE N,et al.Mitochondrial fragmentation affects neither the sensitivity to TNFalpha-induced apoptosis of Brucella-infected cells nor the intracellular replication of the bacteria[J].Scientific Reports,2018,8:5173.
[22] HE Y,REICHOW S,RAMAMOORTHY S,et al.Brucella melitensis triggers time-dependent modulation of apoptosis and down-regulation of mitochondrion-associated gene expression in mouse macrophages[J].Infection & Immunity,2006,74:5035-5046.
[23] MICHAEL O,EZE L Y,ROBERT M,et al.Effects of opsonization and gamma interferon on growth of Brucella melitensis 16M in mouse peritoneal macrophages in vitro[J].Infection and Immunity,2000,74:257-263.
[24] JONES S M,WINTER A J.Survival of virulent and attenuated strains of Brucella abortus in normal and gamma interferon-activated murine peritoneal macrophages[J].Infection and Immunity,1992,60:3011-3014.
[25] CHEN X,LIU H,SUN W,et al.Elevated urine histone 4 levels in women with ovarian endometriosis revealed by discovery and parallel reaction monitoring proteomics[J].Journal of Proteomics,2019,204:103398.
[26] JAIN A P,SAMBATH J,SATHE G,et al.Pan-cancer quantitation of epithelial-mesenchymal transition dynamics using parallel reaction monitoring-based targeted proteomics approach[J].Journal of Translational Medicine,2022,20(1):84.

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 Complex Ⅰ of Mitochondria in Sertoli Cells

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	李婷婷, 刘子超, 周永顺, 何营, 耿方豪, 陈洁然, 高剑峰. 两种处理方法对小鼠骨髓源性树突状细胞体外递呈布鲁氏菌免疫多肽组分析的影响[J]. 中国畜牧兽医, 2023, 50(9): 3730-3739.
[2]	李倩倩, 蔺思函, 王丽媛, 杨兵兵, 张明达, 沈秀丽, 杜志强. 猪种布鲁氏菌TIR结构域蛋白的生物信息学分析、重组表达及蛋白互作研究[J]. 中国畜牧兽医, 2023, 50(8): 3303-3312.
[3]	徐朕宇, 徐锦凤, 邓肖玉, 王月丽, 何金科, 谢珊珊, 王震, 易继海, 王勇, 苗玉和, 崔丽瑾, 陈创夫. 布鲁氏菌外膜囊泡生物信息学分析及免疫原性评价[J]. 中国畜牧兽医, 2023, 50(7): 2906-2914.
[4]	魏春燕, 郭嘉, 朱德馨, 张伟, 朱嘉乐, 邓兴梅, 贾思锋, 刘良波, 张辉. 布鲁氏菌BPE159基因缺失株的构建及BPE159蛋白对细胞自噬因子表达的影响[J]. 中国畜牧兽医, 2023, 50(1): 26-36.
[5]	邓兴梅, 曹树珠, 郭嘉, 朱德馨, 赵天艺, 柴迎锦, 张伟, 史超, 贾思锋, 张辉. 长链非编码RNA Gm35082-202对布鲁氏菌引起的细胞焦亡的影响[J]. 中国畜牧兽医, 2022, 49(9): 3599-3609.
[6]	肖洋洋, 马忠臣, 李芮芮, 陈创夫, 郑炜, 王勇, 王鹏雁. 布鲁氏菌分泌蛋白BspI生物信息学分析及多克隆抗体制备[J]. 中国畜牧兽医, 2022, 49(8): 3112-3121.
[7]	邱润辉, 关飞虎, 王梓行, 郭嘉, 朱德馨, 张伟, 魏春燕, 霍明凯, 孙志华, 张辉. 布鲁氏菌分泌蛋白BMCO基因缺失株的构建及生物学特性研究[J]. 中国畜牧兽医, 2022, 49(5): 1630-1640.
[8]	邱润辉, 关飞虎, 陶婷婷, 李佳, 赵天艺, 邓兴梅, 史超, 孙志华, 张辉. 布鲁氏菌分泌蛋白BPE005的亚细胞定位及与宿主细胞互作蛋白的筛选[J]. 中国畜牧兽医, 2022, 49(4): 1438-1448.
[9]	王月丽, 邵志然, 易继海, 王勇, 王震, 陈创夫. miR-145a-3p通过靶向ATG14调控布鲁氏菌诱导的RAW264.7自噬[J]. 中国畜牧兽医, 2022, 49(3): 1117-1125.
[10]	谢珊珊, 杨琴, 邓肖玉, 易继海, 王震, 陈创夫. 线粒体融合蛋白2基因干扰与过表达对布鲁氏菌诱导巨噬细胞凋亡的影响[J]. 中国畜牧兽医, 2022, 49(1): 283-293.
[11]	杨江流, 乔军伟, 宋青山, 贾芳. 布鲁氏菌S19毒力基因BvfA的原核表达及生物信息学分析[J]. 中国畜牧兽医, 2021, 48(8): 2746-2753.
[12]	代国栋, 闫鸿斌, 李立, 付宝权, 贾万忠. 人兽共患寄生虫病候选疫苗分子筛选方法研究进展[J]. 中国畜牧兽医, 2021, 48(6): 2177-2187.
[13]	李佳, 邓兴梅, 陶婷婷, 朱德馨, 邱润辉, 王梓行, 吴洁, 张辉. 布鲁氏菌侵染过程中has-miR-5196-3p对炎症小体的调控作用[J]. 中国畜牧兽医, 2021, 48(5): 1507-1515.
[14]	宋胜男, 郭嘉, 杨翌承, 桑春丽, 杨梅花, 王远志. 石河子地区宠物犬蜱的种类鉴定及其携带布鲁氏菌的检测[J]. 中国畜牧兽医, 2021, 48(5): 1717-1724.
[15]	张凡, 徐杰, 李刚, 周永顺, 何营, 韩凯, 陈福龙, 高剑峰. 哈萨克羊iNOS基因多态性与布鲁氏菌病的相关性分析[J]. 中国畜牧兽医, 2021, 48(3): 1102-1111.