菌群感应AHLs分子对小鼠肠道菌群多样性影响的分析
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摘要
目的探讨菌群感应信号分子N-酰基高丝氨酸内酯(N-acyl homoserine lactones,AHLs)对小鼠肠道微生物菌群构成的影响,为感染状态下肠道微生态的紊乱补充理论依据。方法 3-oxo-C_(10)-HSL分子经腹腔注射小鼠后,于24 h分别在小鼠十二指肠及空回肠部位用拭子涂抹法取样,并通过Illumina Hiseq 2500高通量测序平台对样品16S rDNA测序,并进行微生物菌群分类构成及多样性分析。结果在实验小鼠肠道内共检出22个细菌门,109个细菌属;对照组和3-oxo-C_(10)-HSL处理组小鼠肠道不同部位菌群均以厚壁菌门(>50%)和变形菌门(>20%)为主,但两组在属水平菌群构成有明显差异,对照组优势菌属依次为葡萄球菌、埃希氏-志贺菌、乳球菌、假单胞菌、肠球菌,3-oxo-C_(10)-HSL处理组优势菌属主要为肠球菌、埃希氏-志贺菌、葡萄球菌;多样性和丰度分析提示3-oxo-C_(10)-HSL处理组肠道菌群多样性均有提高,大肠埃希菌属等主要优势菌丰度显著上升,而且空回肠部位的菌落丰度最高。结论 AHLs对小鼠肠道菌群构成影响较小,而对菌群内部某些特定类群结构变化影响较明显。
Objective Study the effection of N-acyl homoserine lactones(AHLs) on gut microbes flora in mice so as to supply the theoretical basis of intestinal microecological disorder under infection status. Method After the mice were intraperitoneal injected with 3-oxo-C_(10)-HSL for 24 h, samples of mice duodenum and jejuno-ileum contents were acquired with swab smear method. The sample 16 S rDNA sequencing was detected by Illumina Hiseq 2 500 high-throughput sequencing platform, and the classification of microbial flora composition and diversity were analyzed. Result There were 22 phyla and 109 genera in all identified in the mice gut, and Firmicutes(>50%) and Proteobacteria(>20%) dominated the microbiota. The control group and 3-oxo-C_(10)-HSL treatment group had obvious difference in the level of flora composition, for the former the dominant genera mainly included Staphylococcus, Escherichia-Shigella, galactococcus, Pseudomonas and Enterococcus, and for the latter the dominant genera mainly included Enterococcus, Escherichia-Shigella, and Staphylococcus. It was also revealed that the 3-oxo-C_(10)-HSL treatment could improve the gut microbial diversity, and the abundance of dominant genera such as Escherichia-Shigella and jejuno-ileum microbiota significantly rose. Conclusion AHLs had little effect on gut microbiota in mice, but some specific microbial taxa would be significantly affected.
Objective Study the effection of N-acyl homoserine lactones(AHLs) on gut microbes flora in mice so as to supply the theoretical basis of intestinal microecological disorder under infection status. Method After the mice were intraperitoneal injected with 3-oxo-C_(10)-HSL for 24 h, samples of mice duodenum and jejuno-ileum contents were acquired with swab smear method. The sample 16 S rDNA sequencing was detected by Illumina Hiseq 2 500 high-throughput sequencing platform, and the classification of microbial flora composition and diversity were analyzed. Result There were 22 phyla and 109 genera in all identified in the mice gut, and Firmicutes(>50%) and Proteobacteria(>20%) dominated the microbiota. The control group and 3-oxo-C_(10)-HSL treatment group had obvious difference in the level of flora composition, for the former the dominant genera mainly included Staphylococcus, Escherichia-Shigella, galactococcus, Pseudomonas and Enterococcus, and for the latter the dominant genera mainly included Enterococcus, Escherichia-Shigella, and Staphylococcus. It was also revealed that the 3-oxo-C_(10)-HSL treatment could improve the gut microbial diversity, and the abundance of dominant genera such as Escherichia-Shigella and jejuno-ileum microbiota significantly rose. Conclusion AHLs had little effect on gut microbiota in mice, but some specific microbial taxa would be significantly affected.
引文
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[5] Edgar R C.Search and clustering orders of magnitude faster than BLAST [J].Bioinformatics,2010,26 (19):2460-2461.
[6] Wang Q,Garrity G M,Tiedje J M,et al.Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy [J].Appl Environ Microbiol,2007,73 (16):5261-5267.
[7] Chao A.Non-parametric estimation of the number of classes in a population [J].Scandinavian Journal of Statistics,1984,11:265-270.
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[10] Davis B M,Jensen R,Williams P,et al.The interaction of N-acylhomoserine lactone quorum sensing signaling molecules with biological membranes:implications for inter-kingdom signaling [J].PloS one,2010,5 (10):e13522.
[11] Vikstrom E,Bui L,Konradsson P,et al.The junctional integrity of epithelial cells is modulated by Pseudomonas aeruginosa quorum sensing molecule through phosphorylation-dependent mechanisms [J].Exp Cell Res,2009,315 (2):313-326.
[12] Gardiner S M,Chhabra S R,Harty C,et al.Haemodynamic effects of the bacterial quorum sensing signal molecule,N-(3-oxododecanoyl)-L-homoserine lactone,in conscious,normal and endotoxaemic rats [J].Br J Pharmacol,2001,133 (7):1047-1054.
[13] Campos-Galvao M E,Ribon A O,Araujo E F,et al.Changes in the Salmonella enterica Enteritidis phenotypes in presence of acyl homoserine lactone quorum sensing signals [J].J Basic Microbiol,2016,56 (5):493-501.
[14] Ley R E,Hamady M,Lozupone C,et al.Evolution of mammals and their gut microbes [J].Science,2008,320 (5883):1647-1651.
[15] 郭辉,王翠娥,彭刚,等.VITEK2 Compact全自动微生物分析系统在啮齿类细菌学检测中的应用 [J].实验动物科学,2014,31 (2):16-19.
[16] Redfern J,Verran J.What is a microbiologist?A survey exploring the microbiology workforce [J].FEMS Mmicrobiol Lett,2015,362 (24):fnv208.
[17] Queipo-Ortuno M I,Seoane L M,Murri M,et al.Gut microbiota composition in male rat models under different nutritional status and physical activity and its association with serum leptin and ghrelin levels [J].PloS one,2013,8 (5):e65465.
[18] Li Y,Han L,Xu M,et al.The Primary Research on the Gut Microbes in KKAy Mice [J].Indian J Microbiol,2014,54 (1):12-19.
[19] Shang Q,Yang G,Wang Y,et al.Illumina-based analysis of the rhizosphere microbial communities associated with healthy and wilted Lanzhou lily (Lilium davidii var.unicolor) plants grown in the field [J].World J Microbiol Biotechnol,2016,32 (6):95.
[20] Peng X,Yu K Q,Deng G H,et al.Comparison of direct boiling method with commercial kits for extracting fecal microbiome DNA by Illumina sequencing of 16S rRNA tags [J].J Microbiol Methods,2013,95 (3):455-462.
[21] Swift S,Karlyshev A V,Fish L,et al.Quorum sensing in Aeromonas hydrophila and Aeromonas salmonicida:identification of the LuxRI homologs AhyRI and AsaRI and their cognate N-acylhomoserine lactone signal molecules [J].J Bacteriol,1997,179 (17):5271-5281.
[22] Biswa P,Doble M.Production of acylated homoserine lactone by gram-positive bacteria isolated from marine water [J].FEMS Microbiol Lett,2013,343 (1):34-41.
[23] Winzer K,Falconer C,Garber N C,et al.The Pseudomonas aeruginosa lectins PA-IL and PA-IIL are controlled by quorum sensing and by RpoS [J].J Bacteriol,2000,182 (22):6401-6411.