16S rRNA基因测序技术分析肝阳上亢型脑出血患者的肠道菌群结构

李裕思; 许华冲; 王俊月; 郑凡超; 邓力; 黎敬波; 陈龙娇; 袁淮涛; 陈孝银

doi:10.13422/j.cnki.syfjx.20190550

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16S rRNA基因测序技术分析肝阳上亢型脑出血患者的肠道菌群结构

临床 | 更新时间：2021-02-09

- 16S rRNA基因测序技术分析肝阳上亢型脑出血患者的肠道菌群结构
- Analysis of Bacterial Flora Structure of Patients with Cerebral Hemorrhage Due to Hyperactivity of Liver-Yang by 16S rRNA Gene Sequencing Technique
- 中国实验方剂学杂志 2019年25卷第8期页码：83-88
- 作者机构：
  
  1.广州中医药大学　基础医学院，广州　 510000；
  2.暨南大学　中医学院，广州　 510000；
  3.广州中医药大学　第二临床医学院，广州　 510000；
  4.广州中医药大学　第一附属医院，广州　 510000
- 作者简介：
  
  李裕思，在读硕士，从事五运六气与外感病证证候规律研究，Tel: 020-39358006，E-mail: 2445668577@qq.com
  陈孝银，博士，教授，从事中医外感病因学研究，Tel: 020-85226197，E-mail: tchenxiaoyin@jnu.edu.cn
- 基金信息：
  
  国家自然科学基金项目(81830114;81774164;81473557);广东省自然科学基金项目(2017A030313737)
- DOI：10.13422/j.cnki.syfjx.20190550
  中图分类号： R22;R24;R28;C37;R378.2
- 收稿日期：2018-10-15，
  
  网络出版日期：2018-11-20，
  
  纸质出版日期：2019-04-20
- 稿件说明：
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李裕思, 许华冲, 王俊月, 等. 16S rRNA基因测序技术分析肝阳上亢型脑出血患者的肠道菌群结构[J]. 中国实验方剂学杂志, 2019,25(8):83-88.

Yu-si LI, Hua-chong XU, Jun-yue WANG, et al. Analysis of Bacterial Flora Structure of Patients with Cerebral Hemorrhage Due to Hyperactivity of Liver-Yang by 16S rRNA Gene Sequencing Technique[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(8): 83-88.
李裕思, 许华冲, 王俊月, 等. 16S rRNA基因测序技术分析肝阳上亢型脑出血患者的肠道菌群结构[J]. 中国实验方剂学杂志, 2019,25(8):83-88. DOI： 10.13422/j.cnki.syfjx.20190550.

Yu-si LI, Hua-chong XU, Jun-yue WANG, et al. Analysis of Bacterial Flora Structure of Patients with Cerebral Hemorrhage Due to Hyperactivity of Liver-Yang by 16S rRNA Gene Sequencing Technique[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(8): 83-88. DOI： 10.13422/j.cnki.syfjx.20190550.

摘要

目的：

探讨肝阳上亢型急性脑出血患者与健康人群的肠道菌群差异。

方法：

选取2018年于广州中医药大学第一附属医院颅脑科病房收治的9例肝阳上亢型急性脑出血患者的粪便样本为观察组，同时，选取体检的6例健康人粪便样本为对照组，提取两组样本中细菌总DNA，根据16S rRNA V4区设计引物进行扩增，通过Illumina MiSeq平台进行双端测序(paired-end)，通过生物信息分析软件对测序结果进行分析，比较两组样本菌群组成和结构。

结果：

操作分类单元(OTU)的韦恩图分析(Venn分析)显示观察组与对照组之间OTU数目存在明显差异。偏最小二乘法-判别分析(PLS-DA)显示肝阳上亢型急性脑出血患者与健康人在肠道菌群组成上具有明显差异。物种及其丰度分析上，门分类水平上观察组厚壁菌门与拟杆菌门的相对丰度比值(F/B)较正常组明显增加，其中疣微菌门的相对丰度明显低于对照组(

0.05)。属分类水平上，两组的普雷沃氏菌属、拟杆菌属、阿克曼氏菌属、布劳特氏菌属、氨基酸球菌属相对丰度差异有统计学意义(

0.05，

0.01)，其中观察组拟杆菌属和普雷沃氏菌属的相对丰度比值(B/P)显著高于对照组。种分类水平上两组在普氏菌、阿克曼粘细菌、卵形拟杆菌、脆弱拟杆菌、伶俐瘤胃球菌上具有明显差异(

0.05，

0.01)。主坐标分析显示两组样本被显著分开，说明两组样本菌群结构存在显著差异。

结论：

肝阳上亢型急性脑出血与肠道菌群结构紊乱有关，其中普氏菌和阿克曼粘细菌的相对丰度下降是其发生的主要原因。

Abstract

Objective:

To explore the difference of intestinal flora between the patients with acute cerebral hemorrhage due to hyperactivity of liver-Yang and the healthy population.

Method:

The fecal samples of 9 patients with acute cerebral hemorrhage due to hyperactivity of liver-Yang from the first affiliated hospital of Guangzhou university of traditional Chinese medicine in 2018 were selected as observation group

and 6 stool samples from healthy subjects were selected as the control group.The total bacterial DNA was extracted from the two groups of samples

amplified according to the 16S rRNA V4 region

and paired-end sequencing was performed on the Illumina MiSeq platform.The sequencing results were analyzed by bioinformatics analysis software.The flora composition and structure of the samples from two groups were compared.

Result:

Venn analysis of operational taxonomic units(OTU) showed significant difference in OTU numbers between the observation group and control group.Partial least squares-discriminant analysis(PLS-DA) showed that there was a significant difference in the composition of intestinal flora between patients with acute cerebral hemorrhage and healthy subjects.On the analysis of species and abundance

at the classification level of phylum

compared with the control group

the ratio of relative abundance values of Firmicutes and Bacteroidetes(F/B) in the observation group was significantly increased

and the relative abundance of Verrucomicrobia was significantly decreased(

0.05); at the classification level of genus

there were significant differences between the two groups in the

Prevotella

Bacteroides

Akkermansia

Blautia

and

Acidaminococcus

(

0.05

0.01)

the ratio of relative abundance values of

Bacteroides

and

Prevotella

(B/P) in the observation group was significantly higher than that of the control group; at the classification level of species

there were significant differences between the two groups in

copri

muciniphila

ovatus

fragilis

and

Ruminococcus callidus

(

0.05

0.01). Principal coordinate analysis showed that the two groups of samples were significantly separated

indicating that there were significant differences in flora structure between the two groups.

Conclusion:

Acute cerebral hemorrhage due to hyperactivity of liver-Yang is associated with structural disorder of intestinal flora

which is closely related to the decrease in relative abundance of

copri

and

muciniphila

关键词

Keywords

references

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