基于UPLC-QTOF-MS的单增李斯特菌复方新诺明耐药性菌株代谢组学分析  

Metabolomics Analysis of Trimethoprim-sulfamethoxazole Resistant Strains of Listeria monocytogenes Based on Ultra-high Performance Liquid Chromatography Coupled with Quadrupole Time-of-Flight Mass Spectrometry

作　　者：周继福 王娉[1] 郭佳 陈颖[1] Zhou Jifu;Wang Ping;Guo Jia;Chen Ying(Chinese Academy of Inspection and Quarantine,Beijing 100176;College of Food Science and Engineering,Nanjing University of Finance and Economics,Nanjing 210023;College of Food Science and Engineering,Tianjin University of Science&Technology,Tianjin 300457)

机构地区：[1]中国检验检疫科学研究院,北京100176 [2]南京财经大学食品科学与工程学院,南京210023 [3]天津科技大学食品科学与工程学院,天津300457

出　　处：《中国食品学报》2022年第1期215-226,共12页Journal of Chinese Institute Of Food Science and Technology

基　　金：“十三五”国家重点研发计划重点专项(2018YFC1603800)。

摘　　要：采用非靶标代谢组学方法,研究单增李斯特菌复方新诺明耐药株和敏感株的代谢组学差异。以复方新诺明(SXT)耐药株和敏感株为研究对象,使用超高效液相色谱-四极杆飞行时间质谱联用技术(UPLC-QTOF-MS)获得菌株胞内代谢轮廓信息,建立OPLS-DA判别模型,并对差异化合物的数量、种类及其在代谢通路中的调控表达进行分析。结果表明:耐药株和敏感株的代谢特征存在显著差异,共鉴定出21种对分类有显著贡献的化合物,主要以氨基酸和核苷酸为主。差异代谢物主要参与氨基酸代谢、碳水化合物代谢和核苷酸代谢等通路。腺苷、色氨酸和亮氨酸在2条显著性差异代谢途径中的表达变化表明:菌株受复方新诺明作用后,细胞内的肽、蛋白质合成受阻,细胞氧化应激提高。甘油磷脂代谢表明:细胞调控转运、蛋白功能和信号转导受复方新诺明严重影响。本研究阐明了菌株受复方新诺明作用的耐药性形成原因,为药物作用靶点的寻找和干预菌株防控提供帮助。In this study,non-target metabolomics methods were used to study the metabonomic differences between trimethoprim-sulfamethoxazole(SXT)-resistant and sensitive strains of Listeria monocytogenes.Ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-QTOF-MS) was used to obtain the intracellular metabolic profile information of the strains contained by SXT-resistant and sensitive strains.OPLS-DA discriminant models were established to illustrate the metabonomics differences,then the number and type of different compounds were qualitatived and their regulatory expression in metabolic pathways were analyzed.The results showed that there were significant differences in the metabolic characteristics of drug-resistant strains and sensitive strains.A total of 21 compounds that contributed significantly to the classification were identified,mainly amino acids and nucleotides.Differential metabolites are mainly involved in the pathways of amino acid metabolism,carbohydrate metabolism and nucleotide metabolism.The expression changes of adenosine,tryptophan and leucine in two significantly different metabolic pathways indicate that the synthesis of peptides and proteins in the cell has been blocked and the cell oxidative stress has increased after the strain is affected by the compound trimethoprim.The metabolism of glycerophospholipids indicated that the cell regulation of transport,protein function,and signal transduction were severely affected by the strains being affected by the SXT.The study clarified the reasons for the formation of the drug resistance of the strains affected by the SXT,and provided help for the search for drug targets and intervention in the prevention and control of strains.

关 键 词：单增李斯特菌 耐药性 非靶标代谢组学 代谢通路 

分 类 号：O657.63[理学—分析化学] TS201.3[理学—化学]