感染减毒鼠伤寒沙门氏菌对小鼠粪样代谢组的影响——WIPM 和Bruker 500 MHz 核磁共振波谱仪检测结果的比较

波谱学杂志

感染减毒鼠伤寒沙门氏菌对小鼠粪样代谢组的影响——WIPM 和Bruker 500 MHz 核磁共振波谱仪检测结果的比较

陈璐1,2，宋侃 1,3，王玉兰1*

1. 中国科学院生物磁共振分析重点实验室，波谱与原子分子物理国家重点实验室，武汉磁共振中心（中国科学院武汉物理与数学研究所），湖北武汉 430071；2. 长江航务管理局疾病预防控制中心，湖北武汉 430019；3. 中国科学院大学，北京100049

收稿日期:2014-01-14 修回日期:2014-07-21 出版日期:2014-09-05 发布日期:2014-09-05
作者简介:陈璐(1988-)，女，江西抚州人，硕士研究生，主要从事生物医学代谢组学研究，E-mail：tjchenlu@163.com. *通讯联系人：王玉兰，电话：027-87198430，E-mail：yulan.wang@wipm.ac.cn.
基金资助:
国家重大科学仪器设备开发专项资助项目(2011YQ12003503).

Effects of Attenuated Salmonella Typhimurium Infection on Fecal Metabonome in Mice——comparison between WIPM and Bruker 500 MHz NMR spectrometers

CHEN Lu1,2，SONG Kan1,3，WANG Yu-lan1*

1. Key Laboratory of Magnetic Resonance in Biological Systems, State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, National Center for Magnetic Resonance in Wuhan (Wuhan Institute of Physics and Mathematics, Chinese Academy of Sciences), Wuhan 430071, China;
2. Center for Disease Prevention and Control of Changjiang River Administration of Navigational Affairs, Wuhan 430019, China;
3. University of Chinese Academy of Sciences, Beijing 100049, China

Received:2014-01-14 Revised:2014-07-21 Online:2014-09-05 Published:2014-09-05
About author:WANG Yu-lan, Tel: +86-27-87197143, E-mail: yulan.wang@wipm.ac.cn.
Supported by:
国家重大科学仪器设备开发专项资助项目(2011YQ12003503).

摘要/Abstract

摘要：

鼠伤寒沙门氏菌属于胞内侵袭性兼性厌氧菌，减毒后的鼠伤寒沙门氏菌安全性大大提高，已被广泛用于病毒、细菌及寄生虫疫苗的研制．但是目前还没有研究利用代谢组学方法来分析减毒鼠伤寒沙门氏菌感染对宿主和肠道菌群相互作用的影响．同时中国科学院武汉物理与数学研究所自主研发的WIPM 500 MHz 核磁共振(NMR)波谱仪已经研制成功，但是该谱仪是否可以用于代谢组学研究尚未经过考证．本研究同时使用WIPM 和Bruker500 MHz 核磁共振波谱仪对同一批减毒鼠伤寒沙门氏菌感染小鼠的粪样进行检测，并结合多变量数据分析方法来研究减毒鼠伤寒沙门氏菌感染对小鼠粪样代谢组的影响．研究结果表明，WIPM NMR 波谱仪检测数据的分析结果与Bruker NMR 波谱仪的实验结果具有高度的一致性，表明WIPM 谱仪可以用于代谢组学研究．研究发现减毒鼠伤寒沙门氏菌感染能够引起小鼠粪样代谢物中多种氨基酸和尿嘧啶含量的上升，并伴有胆汁酸、乳酸和丙酸含量的下降．以上代谢物含量的改变表明，减毒鼠伤寒沙门氏菌感染使小鼠肠道微生物的代谢功能发生了紊乱．

关键词: 核磁共振(NMR), 粪样代谢组, 减毒鼠伤寒沙门氏菌, 谱仪对比

Abstract:

Salmonella typhimurium is a intracellular, invasive, facultative anaerobic bacteria. With improved safety, attenuated salmonella typhimurium has been widely used in development of vaccine for preventing viruses, bacteria, and parasites infections. However, limited research has been done so far on the biological effects of attenuated salmonella typhimurium infection on animal hosts. In this work, we measured fecal metabonome of mice infected with attenuated salmonella typhimurium using NMR spectroscopy. Two sets of NMR data were acquired with the same protocols on a Bruker 500 MHz NMR spectrometer and an in-house 500 MHz NMR spectrometer developed in Wuhan Institute of Physics and Mathematics (WIPM). Multivariate data analysis methods were used to analyzed the data. It was found that infection induced increases of certain amino acids and uracil in feces, and decreases of bile acids, lactate and propionate. Such metabolic alterations indicated that infection of attenuated salmonella typhimurium may cause disturbance of gut microbiotal communities. The results obtained from the home-developed spectrometer in WIPM showed high consistency with those obtained from Bruker 500 MHz NMR spectrometer, suggesting that the WIPM spectrometer is ready for metabonomic studies.

Key words: NMR, fecal metabonome, attenuated salmonella typhimurium, spectrometer

中图分类号:

O482.53

陈璐1,2，宋侃 1,3，王玉兰1*. 感染减毒鼠伤寒沙门氏菌对小鼠粪样代谢组的影响——WIPM 和Bruker 500 MHz 核磁共振波谱仪检测结果的比较[J]. 波谱学杂志.

CHEN Lu1,2，SONG Kan1,3，WANG Yu-lan1*. Effects of Attenuated Salmonella Typhimurium Infection on Fecal Metabonome in Mice——comparison between WIPM and Bruker 500 MHz NMR spectrometers[J]. Chinese Journal of Magnetic Resonance.

参考文献

[1] Everest P, Ketley J, Hardy S, et al. Evaluation of salmonella typhimurium mutants in a model of experimental gastroenteritis[J]. Infect Immun, 1999, 67(6): 2 815－2 821.

[2] Hur J, Lee J H. Immune responses to new vaccine candidates constructed by a live attenuated salmonella typhimurium delivery system expressing escherichia coli f4, f5, f6, f41 and intimin adhesin antigens in a murine model[J]. J Vet Med Sci, 2011, 73(10): 1 265－1 273.

[3] Xin W, Li Y, Mo H, et al. Pspa family fusion proteins delivered by attenuated salmonella enterica serovar typhimurium extend and enhance protection against streptococcus pneumoniae[J]. Infect Immun, 2009, 77(10): 4 518－4 528.

[4] Yang X L, Liu W C, Yang W W, et al. Oral immunization of mice with vaccine of attenuated salmonella typhimurium expressing helicobacter pylori urease b subunit[J]. Biomed Environ Sci, 2005, 18(6): 411－418.

[5] Rahman M M, Uyangaa E, Han Y W, et al. Oral administration of live attenuated salmonella enterica serovar typhimurium expressing chicken interferon-alpha alleviates clinical signs caused by respiratory infection with avian influenza virus h9n2[J]. Vet Microbiol, 2011, 154(1-2): 140－151.

[6] Cheng Ming(程明), He Jin-sheng(何金生), Fu Yuan-hui(付远辉), et al. Antibody responses induced by mucosal DNA vaccine encoding the codon-optimized F protein of human respiratory syncytial virus (RSV) delivered with attenuated salmonella typhimurium(以减毒鼠伤寒沙门菌为载体的人呼吸道合胞病毒F 蛋白DNA 疫苗滴鼻免疫抗体分析)[J]. Acta

Microbiologica Sinica(微生物学报), 2011, 51(7): 965－971.

[7] Chen G, Dai Y, Chen J, et al. Oral delivery of the sj23lhd-gst antigen by salmonella typhimurium type iii secretion system protects against schistosoma japonicum infection in mice[J]. PLoS Negl Trop Dis, 2011, 5(9): e1313.

[8] Yang Y, Zhang Z, Yang J, et al. Oral vaccination with ts87 DNA vaccine delivered by attenuated salmonella typhimurium elicits a protective immune response against trichinella spiralis larval challenge[J]. Vaccine, 2010, 28(15): 2 735－2 742.

[9] Qu D, Yu H, Wang S, et al. Induction of protective immunity by multiantigenic DNA vaccine delivered in attenuated salmonella typhimurium against toxoplasma gondii infection in mice[J]. Vet Parasitol, 2009, 166(3-4): 220－227.

[10] Tang H, Wang Y. Metabonomics：A revolution in progress[J]. Prog Biochem Biophys, 2006, 33(5): 401－417.

[11] Chen Lu(陈璐), Wang Yu-lan(王玉兰). Metabolite composition of planarian analyzed by nuclear magnetic resonance spectroscopy (涡虫代谢物组成的核磁共振分析)[J]. Chinese J Magn Reson(波谱学杂志) , 2013, 30(4): 516－527.

[12] Cevallos-Cevallos J M, Danyluk M D, Reyes-De-Corcuera J I. Gc-ms based metabolomics for rapid simultaneous detection of escherichia coli o157:H7, salmonella typhimurium, salmonella muenchen, and salmonella hartford in ground beef and chicken[J]. J Food Sci, 2011, 76(4): M238－M246.

[13] Antunes L C M, Arena E T, Menendez A, et al. Impact of salmonella infection on host hormone metabolism revealed by metabolomics[J]. Infect Immun, 2011, 79(4): 1 759－1 769.

[14] Clairmont C, Lee K C, Pike J, et al. Biodistribution and genetic stability of the novel antitumor agent vnp20009, a genetically modified strain of salmonella typhimurium[J]. J Infect Dis, 2000, 181(6): 1 996－2 002.

[15] Clairmont C, Bermudes D, Ittensohn M, et al. Genetically modified salmonella typhimurium, vnp20009, a novel anticancer agent: Bio-distribution, genetic stability, selective tumor accumulation, and antitumor efficacy[J]. Cancer Gene Ther, 1999, 6(6): S11－S12.

[16] Nicholson J K, Connelly J, Lindon J C, et al. Metabonomics: A platform for studying drug toxicity and gene function[J]. Nat Rev Drug Discov, 2002, 1(2): 153－161.

[17] Tian Yuan(田园), Tang Hui-ru(唐惠儒). Identification and structural determination of saccharides in rat feces(大鼠粪样中几种糖类物质的结构确定)[J]. Chinese J Magn Reson(波谱学杂志) , 2012, 29(3): 361－371.

[18] Jolliffe I. Principal Component Analysis[M]. Wiley Online Library: 2005.

[19] Eriksson L, Trygg J, Wold S. CV-ANOVA for significance testing of pls and opls® models[J]. J Chemometr, 2008, 22(11-12): 594－600.

[20] Jiang L, Huang J, Wang Y, et al. Metabonomic analysis reveals the ccl4-induced systems alterations for multiple rat organs[J]. J Proteome Res, 2012, 11(7): 3 848－3 859.

[21] Fan T W M, Lane A N. Structure-based profiling of metabolites and isotopomers by NMR[J]. Prog Nucl Mag Res Sp, 2008, 52(2-3): 69－117.

[22] Wang Y, Tang H, Holmes E, et al. Biochemical characterization of rat intestine development using high-resolution magic-angle-spinning 1h nmr spectroscopy and multivariate data analysis[J]. J Proteome Res, 2005, 4(4): 1 324－1 329.

[23] Marchesi J R, Holmes E, Khan F, et al. Rapid and noninvasive metabonomic characterization of inflammatory bowel disease[J]. J Proteome Res, 2007, 6(2): 546－551.

[24] Johnston S J, Ridge S A, Cassidy J, et al. Regulation of dihydropyrimidine dehydrogenase in colorectal cancer[J]. Clin Cancer Res, 1999, 5(9): 2 566－2 570.

[25] Jones B V, Begley M, Hill C, et al. Functional and comparative metagenomic analysis of bile salt hydrolase activity in the human gut microbiome[J]. Proc Natl Acad Sci USA, 2008, 105(36): 13 580－13 585.

[26] Grubb B R. Avian cecum: Role of glucose and volatile fatty acids in transepithelial ion transport[J]. Am J Physiol-Gastr L, 1991, 260(5): G703－G710.

[27] Gill S R, Pop M, DeBoy R T, et al. Metagenomic analysis of the human distal gut microbiome[J]. Science, 2006, 312(5778):