基于高通量测序的老年人肠道菌群多样性分析  

Analysis of gut microbiota diversity in the elderly based on high-throughput sequencing

作　　者：邱惠萍[1,2] 姚水洪 卢伟力[3] 洪萌[2] 任艳丽 邵骏威 陈智 Qiu Huiping;Yao Shuihong;Lu Weili;Hong Meng;Ren Yanli;Shao Junwei;Chen Zhi(Department of Infectious Diseases,Medical School,Quzhou College of Technology,Quzhou 324000,China;State Key Laboratory for Diagnosis and Treatment of Infectious Diseases,the First Affiliated Hospital,College of Medicine,Zhejiang University,Hangzhou 310003,China;Department of Infectious Diseases,People′s Hospital of Quzhou,Quzhou 324000,China)

机构地区：[1]衢州职业技术学院医学院传染病学教研室,324000 [2]浙江大学医学院附属第一医院传染病诊治国家重点实验室,杭州310003 [3]衢州市人民医院感染科,324000

出　　处：《中华微生物学和免疫学杂志》2020年第4期262-268,共7页Chinese Journal of Microbiology and Immunology

基　　金：十三五国家科技重大专项(2018ZX10302206);浙江省教育厅高校访问学者教师专业发展项目(FG2018158);衢州市第三批科技重点创新团队项目(201706)。

摘　　要：目的:利用高通量测序技术,研究老年人肠道菌群多样性随年龄变化的特点。方法:招募90例健康志愿者为研究对象,按年龄进行分组,其中<60岁(中青年组)为基线对照Age A组,年龄≥60岁者以每10岁为一个年龄段分为Age B、Age C、Age D、Age E 4组。采集研究对象粪便样本,提取DNA,采用二代高通量基因测序技术,对16S rDNA V3~V4高变区进行扩增测序、生物信息学分析,比较不同年龄老年人群肠道菌群和功能基因的差异。结果:在门水平上,不同年龄组肠道菌群组成包括厚壁菌门、拟杆菌门、变形菌门、放线菌门4个主要门,其中占比最高的是厚壁菌门,占60%以上,其次为拟杆菌门;从属水平上,粪杆菌属(Faecalibacterium)随着年龄增加丰度不断降低。α多样性分析显示,不同年龄老年人肠道菌群的丰度较高,均匀度较好,各组间无明显差异性;但是β多样性分析发现,Age A组和Age B组群落结构组成差异较大;Age B组和Age C组群落结构相似性最高。结论:从中青年过渡到60岁以上,肠道菌群的群落结构发生了变化,在60~80岁趋于相对稳定,年龄超过80岁其菌群结构再次发生变化,老年人慢性炎症性疾病、代谢性疾病和肿瘤多发可能与Faecalibacterium菌的降低有关。Objective To investigate the changes in gut microbiota diversity with age in elderly people using high-throughput sequencing.Methods Ninety healthy volunteers were recruited.People who were<60 years old(middle-aged group)were set up as a baseline control group(Age A group),while those aged≥60 years old were further divided into four groups(60-<70:Age B group,70-<80:Age C group,80-<90:Age D group,≥90:Age E group).Fecal samples were collected to extract DNA.The second-generation sequencing technology was used to amplify and sequence the V3-V4 hypervariable region of 16S rDNA.Bioinformatics analysis was performed to compare the differences in gut microbiota and functional genes among groups.Results At the phylum level,gut microbiota were composed mainly of Firmicute,Bacteroidetes,Proteobacteria and Actinobacteria in different groups.The proportion of Firmicute was the highest,accounting for over 60%,followed by that of Bacteroidetes.At the genus level,the abundance of Faecalibacterium genus decreased with age.Theαdiversity analysis showed that the gut microbiota in the elderly of different ages had higher abundance and uniformity,and there was no significant difference among groups.However,theβdiversity analysis showed that in community structure there was difference between Age A and Age B groups,and similarity between Age B and Age C groups.Conclusions The community structure of gut microbiota changed significantly between young and middle-aged people and the elderly over 60 years old.It tended to be relatively stable in people of 60-80 years old,but changed again when they were over 80 years old.Chronic inflammatory diseases,metabolic diseases and tumors in the elderly might be associated with the decrease in Faecalibacterium.

关 键 词：老年人 肠道菌群 高通量测序 PICRUSt功能预测 

分 类 号：R3[医药卫生—基础医学]