应用454测序技术评价非酒精性脂肪性肝病患者肠道菌群结构差异*

doi:10.3969/j.issn.1672-5069.2015.02.010

实用肝脏病杂志 ›› 2015, Vol. 18 ›› Issue (2): 145-149.doi: 10.3969/j.issn.1672-5069.2015.02.010

应用454测序技术评价非酒精性脂肪性肝病患者肠道菌群结构差异^*

沈峰, 陈建能, 郑瑞丹, 王晓颖, 潘勤, 陈光榆, 章瑞南, 徐雷鸣, 范建高

200092 上海市上海交通大学医学院附属新华医院消化内科（沈峰,潘勤,章瑞南,徐雷鸣,范建高）; 漳州市正兴医院（陈建能,郑瑞丹）; 上海交通大学医学院附属新华医院病理科（王晓颖）; 上海交通大学医学院临床流行病学研究中心（陈光榆）

收稿日期:2014-10-09 出版日期:2015-03-10 发布日期:2016-02-19
通讯作者: 范建高,E-mail:fanjiangao@gmail.com
作者简介:沈峰,男,34岁,医学博士,主治医师。E-mail：feng-shen@hotmail.com;陈建能,男,27岁,医学硕士。E-mail：chenjn0596@163.com
基金资助:
国家重点基础研究发展计划（973）项目（编号：2012CB517501）; 中国肝炎防治基金会“天晴肝病”研究基金（编号：20120027）

Constitutional variation of gut microbiota detected by bar coded 454 pyrosequencing technology in patients with non-alcoholic fatty liver diseases

Shen Feng, Chen Jianneng, Zheng Ruidan, et al.

Department of Gastroenterology,Xinhua Hospital,Shanghai Jiaotong University School of Medicine,Shanghai 200092,China

Received:2014-10-09 Online:2015-03-10 Published:2016-02-19

摘要/Abstract

摘要： 目的阐明非酒精性脂肪性肝病(NAFLD)患者肠道菌群结构特征。方法采用454焦磷酸测序技术,对47例经“肝活检”确诊的NAFLD患者和34例健康人新鲜粪便样品16S DNA V3~V5可变区进行测序,通过序列比较及操作分类单元（OTU）划分评价肠道菌群结构差异。结果 NAFLD患者OTUs为（88.32±28.27）,显著低于健康人[（109.65±30.65）,P<0.01];在“门”水平,NAFLD患者厚壁菌门所占比例为（51.2±17.8）%,显著高于健康人的[（46.4±12.8）%,P=0.048],而拟杆菌门为（31.6±18.9）%,显著低于健康人的[（43.3±14.4）%,P<0.001];在“纲”水平上,NAFLD组Erysipelotrichi纲占（3.2±5.1）%,显著高于健康人的[（1.0±1.2）%,P=0.009],而Bacteroidia纲占（31.0±18.8）%,显著低于对照组的[（42.3±14.0）%,P=0.004];在“属”水平上,NAFLD组乳球菌占（0.0038±0.0001）%,低于健康组的[（0.0145±0.001）%,P=0.003],普氏菌属也同样显著减少（P=0.022）,而链球菌（Streptococcus）在NAFLD占（1.50±0.03）%,显著高于对照组的[（0.21±0.24）%,P=0.004]。结论 NAFLD患者肠道菌群多样性减少,存在构成显著异常,是致病因素抑或其后果仍有待于进一步研究。

关键词: 非酒精性脂肪性肝病, 肠道菌群, 454焦磷酸测序

Abstract: Objective To elucidate the changes of gut microbiota projects in patients with non-alcoholic fatty liver diseases（NAFLD）. Methods Fresh faecal samples from 47 NAFLD patients underwent liver biopsies and 34 health individuals were examined using bar coded 454 pyrosequencing technology. 16S DNA V3~V5 variable regions were sequenced and the operational taxonomy unit （OUT） was analyzed to evaluate the differences of microbiota projects. Results The OUT of patients with NAFLD were（88.32±28.27）,significantly lower than those in the controls[（109.65±30.65）,P<0.01）];In Phylum level,the abundance of Firmicutes had significantly increased in NAFLD patients than in the controls[（51.2±17.8）% vs.（46.4±12.8）%,P=0.048],while the abundance of Bacteroidetes had significantly reduced[（31.0±18.8）% vs.（43.3±14.4）%,P<0.01];In Class level,the abundance of Erysipelotrichi had significantly increased in NAFLD patients than in the controls [（3.2±5.1）% vs. （1.0±1.2）%,P=0.009],while the abundance of Bacteroidia had significantly decreased [（31.0±18.8）% vs.（42.3±14.0）%,P=0.004]; In Genus level, the Lactococcus[（0.0038±0.0001）%] had significantly reduced in NAFLD patients compared with controls [（0.0145±0.001）%,P=0.003];The same results were found in Prevotella Genus. However,Streptococcus had significantly increased in NAFLD patients [（1.50±0.03）%] than in controls （0.21±0.24）%,P=0.004]. Conclusions Patients with NAFLD had a lower diversity of gut microbiota projects.

Key words: Non-alcoholic fatty liver disease, Gut microbiota, Bar coded 454 pyrosequencing

沈峰, 陈建能, 郑瑞丹, 王晓颖, 潘勤, 陈光榆, 章瑞南, 徐雷鸣, 范建高. 应用454测序技术评价非酒精性脂肪性肝病患者肠道菌群结构差异^*[J]. 实用肝脏病杂志, 2015, 18(2): 145-149.

Shen Feng, Chen Jianneng, Zheng Ruidan, et al.. Constitutional variation of gut microbiota detected by bar coded 454 pyrosequencing technology in patients with non-alcoholic fatty liver diseases[J]. JOURNAL OF PRACTICAL HEPATOLOGY, 2015, 18(2): 145-149.

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应用454测序技术评价非酒精性脂肪性肝病患者肠道菌群结构差异^*

Constitutional variation of gut microbiota detected by bar coded 454 pyrosequencing technology in patients with non-alcoholic fatty liver diseases

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