基于16SrDNA测序的肥胖儿童肠道菌群变化分析
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摘要
目的通过16SrDNA测序研究肥胖儿童肠道微生物的组成及变化特点,为后期相关研究奠定基础。方法于2017年4月-2018年4月选取肥胖儿童及正常体重儿童共59例,收集粪便,采用16SrDNA扩增子测序的方法检测儿童肠道菌群组成。结果肥胖组儿童和正常体重组儿童的肠道微生物多样性差异无统计学意义(P>0.05),但分布规律和数量差异有统计学意义(P<0.05)。在门水平中厚壁菌门、拟杆菌门、放线菌门是儿童肠道微生物群落中最丰富的菌群。其中疣微菌门数量在正常体重组显著高于肥胖组儿童。在属水平,发现多尔氏菌数量在肥胖组儿童肠道内显著高于正常体重组儿童,嗜黏蛋白阿克曼氏菌数量在正常体重组儿童显著高于肥胖组儿童。结论肠道菌群分布规律和数量的改变与肥胖的发生有密切联系。
Objective To study the composition and characteristics of intestinal microorganisms in obese children by illumina amplicon sequencing of 16 SrDNA Tag,in order to provide basis for further related research. Methods Totally 59 obese children and children with normal weight were enrolled in this study,and their feces were collected.16 SrDNA method was used to detect the composition and difference of intestinal flora in different groups. Results There was no significant difference on intestinal microbial diversity between obese and normal weight children(P>0.05),but significant differences on distribution and quantity were found(P<0.05).At the phylum level,Firmicutes,Bacteroidetes and Acidobacteria were the most abundant flora in children′s intestinal microbial community.The number of Verrucomicrobia in normal weight group was significantly higher than that in obese group.At the genus level,it was found that the number of Dorea was significantly higher in obese group than that in normal weight group,while the number of Akkmermansia was significantly higher in normal weight children. Conclusion The change of intestinal flora distribution pattern and quantity is closely related to the occurrence of obesity.
Objective To study the composition and characteristics of intestinal microorganisms in obese children by illumina amplicon sequencing of 16 SrDNA Tag,in order to provide basis for further related research. Methods Totally 59 obese children and children with normal weight were enrolled in this study,and their feces were collected.16 SrDNA method was used to detect the composition and difference of intestinal flora in different groups. Results There was no significant difference on intestinal microbial diversity between obese and normal weight children(P>0.05),but significant differences on distribution and quantity were found(P<0.05).At the phylum level,Firmicutes,Bacteroidetes and Acidobacteria were the most abundant flora in children′s intestinal microbial community.The number of Verrucomicrobia in normal weight group was significantly higher than that in obese group.At the genus level,it was found that the number of Dorea was significantly higher in obese group than that in normal weight group,while the number of Akkmermansia was significantly higher in normal weight children. Conclusion The change of intestinal flora distribution pattern and quantity is closely related to the occurrence of obesity.
引文
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[3] Fullston T,Ohlsson Teague EM,Palmer NO,et al.Paternal obesity initiates metabolic disturbances in two generations of mice with incomplete penetrance to the F2generation and alters the transcriptional profile of testis and sperm microRNA content[J].FASEB J,2013,27(10):4226-4243.
[4] Marchesi JR.Prokaryotic and eukaryotic diversity of the human gut[J].Adv Appl Microbiol,2010,72:43-62.
[5]李辉,季成叶,宗心南,等.中国0~18岁儿童、青少年体块指数的生长曲线[J].中华儿科杂志,2009,47(7):493-498.
[6] Archer SD,McDonald IR,Herbold CW,et al.Benthic microbial communities of coastal terrestrial and ice shelf Antarctic meltwater ponds[J].Front Microbiol,2015,6:485.
[7] Edgar RC.UPARSE:highly accurate OTU sequences from microbialamplicon reads[J].Nat Methods,2013,10(10):996-998.
[8] Wang Q,Garrity GM,Tiedje JM,et al.Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy[J].AEM,2007,16(73):5261-5267.
[9] Quast C,Pruesse E,Yilmaz P,et al.The SILVA ribosomal RNA gene database project:improved data processing and web-based tools[J].Nucleic.Acids Res,2013,41(D1):590-596.
[10]李东萍,郭明璋,许文涛.16SrRNA测序技术在肠道微生物中的应用研究进展[J].生物技术通报,2015,31(2):71-77.
[11] Karlsson CL,nnerflt J,Xu J,et al.The microbiota of the gut in preschool children with normal and excessive body weight[J].Obesity,2012,20(11):2257-2261.
[12]李静,路媛媛,樊超男,等.学龄前肥胖儿童肠道菌群的变化[J].中国儿童保健杂志,2015,23(4):388-390.
[13] Xu P,Li M,Zhang J,et al.Correlation of intestinal microbiota with overweight and obesity in Kazakh school children[J].BMC Microbiol,2012,12:283.
[14] Duburcq T,Hubert T,Saint-Leger P,et al.Impact of endotoxin challenge in obese pigs[J].Shock,2014,41(6):546-553.
[15] Jeon W,Kim YC,Hong M,et al.Microcrystalline cellulose for delivery of recombinant protein-based antigen against erysipelas in mice[J].Biomed Res Int,2018,2018:7670505.
[16] Denes E,Camilleri Y,Fiorenza F,et al.First case of osteomyelitis due to Erysipelothrix rhusiopathiae:pubic osteomyelitis in a gored farmer[J].Int J Infect Dis,2015,30:133-134.
[17] Zitak M,Kovatcheva-Datchary P,Markiewicz,LH,et al.Altered microbiota contributes to reduced diet-induced obesity upon cold exposure[J].Cell Metab,2016,23(6):1216-1223.
[18] Flint HJ,Rincon EA,Marco MT,et al.Polysaccharide utilization by gut bacteria:potential for new insights from genomic analysis[J].Nat Rev Microbiol,2008,6(2):121-131.
[19] Verdam FJ,Fuentes S,de Jonge C,et al.Human intestinal microbiota composition is associated with local and systemic inflammation in obesity[J].Obesity,2013,21(12):E607-615.
[20] Samuel BS,Shaito A,Motoike T,et al.Effects of the gut microbiota on host adiposity are modulated by the shortchain fatty-acid binding G protein-coupled receptor,Gpr41[J].Proc Natl Acad Sci USA,2008,105(43):167.