基于16S rRNA技术分析石珍安神汤对脱髓鞘小鼠肠道菌群多样性的影响

doi:10.3969/j.issn.1006-7795.2020.01.013

首都医科大学学报 ›› 2020, Vol. 41 ›› Issue (1): 64-70.doi: 10.3969/j.issn.1006-7795.2020.01.013

• 精神疾病中西医结合治疗 • 上一篇下一篇

基于16S rRNA技术分析石珍安神汤对脱髓鞘小鼠肠道菌群多样性的影响

马超^1,2, 贺毅^1,2, 孙作厘^1,2, 刘鑫垚^1,2, 冯正田^1,2, 陈沛^1,2, 宁艳哲^1,2, 朱虹^1,2, 尹冬青^1,2, 贾竑晓^1,2

1. 首都医科大学附属北京安定医院国家精神心理疾病临床研究中心精神疾病诊断与治疗北京市重点实验室, 北京 100088;
2. 首都医科大学人脑保护高精尖创新中心, 北京 100069

收稿日期:2019-12-02 出版日期:2020-02-21 发布日期:2020-02-13
通讯作者: 贾竑晓 E-mail:jhxlj@ccmu.edu.cn
基金资助:
国家自然科学基金项目（81873398，81904120），首都卫生发展科研专项重点攻关项目（2018-1-2122），北京中医药科技发展基金青年项目（QN 2018-27），北京市医院管理中心"登峰"计划（DFL20191901）。

Analysis of effect of Shi-Zhen-An-Shen-Tang on intestinal microflora diversity of mice exposed to cuprizone based on 16S rRNA technique

Ma Chao^1,2, He Yi^1,2, Sun Zuoli^1,2, Liu Xinyao^1,2, Feng Zhengtian^1,2, Chen Pei^1,2, Ning Yanzhe^1,2, Zhu Hong^1,2, Yin Dongqing^1,2, Jia Hongxiao^1,2

1. The National Clinical Research Center for Mental Disorders & Beijing Key Laboratory of Mental Disorders, Beijing Anding Hospital, Capital Medical University, Beijing 100088, China;
2. Advanced Innovation Center for Human Brain Protection, Capital Medical University, Beijing 100069, China

Received:2019-12-02 Online:2020-02-21 Published:2020-02-13
Supported by:
This study was supported by National Natural Science Foundation of China (81873398, 81904120), Capital Health Research and Development of Special Funding (2018-1-2122), Chinese Medicine Science and Technology Development Fund of Beijing (QN 2018-27),Beijing Hospitals Authority's Ascent Plan(DFL20191901).

摘要/Abstract

摘要： 目的基于16S rRNA技术研究石珍安神汤（Shi-Zhen-An-Shen-Tang，SZAST）对脱髓鞘模型小鼠肠道菌群多样性的影响，并探讨该复方治疗精神分裂症的作用机制。方法 SPF级5周龄雄性C57BL/6小鼠30只，采用数字表法随机分为普通饲料对照组、模型组、中药低剂量组、中药中剂量组、中药高剂量组、喹硫平组，每组5只。适应性喂养2周后，对照组正常饮食，其他组食用混合了0.2%（质量分数）双环乙酮草酰二腙（cuprizone，CPZ）的特殊饲料。食用特殊饲料4周后，SZAST组给予SZAST灌胃2周（低浓度组5.5 g·kg^-1·d^-1，中浓度组11.0 g·kg^-1·d^-1，高浓度组16.5 g·kg^-1·d^-1），模型组以0.9%（质量分数）氯化钠注射液灌胃2周（10 mL·kg^-1·d^-1），喹硫平组按10 mg·kg^-1·d^-1给予喹硫平溶液灌胃。2周后小鼠麻醉取盲肠内容物。以Illumina MiSeq为测序平台，对其16S rRNA V3-V4区的肠道菌群可操作分类单元（operational taxonomic units，OTUs）数量，alpha与beta多样性，丰富度指数与多样性指数以及差异菌与菌属进行综合性分析与评价。结果石珍安神汤可改善脱髓鞘小鼠OTUs数量及alpha与beta多样性的失调，对脱髓鞘小鼠的β-变形菌纲、伯克菌目、拟杆菌科、产碱杆菌科、毛螺菌属-NK4A136等差异菌分布结构均产生回调作用。结论石珍安神汤对脱髓鞘模型小鼠的异常菌群多样性起到调节作用，并通过16S rRNA技术揭示了肠道菌群在白质损害导致精神分裂症中的相关病理机制。

关键词: 精神分裂症, 石珍安神汤, 双环己酮草酰二腙, 肠道菌群, 高通量测序, 16S rRNA 技术, 可操作分类单元

Abstract: Objective To explore the effect of Shi-Zhen-An-Shen-Tang(SZAST) on intestinal microflora diversity of demyelinated model mice, and further to reveal its therapeutic characteristics and mechanisms based on the 16S rRNA technique. Methods Thirty Young adult male C57BL/6 mice (5 weeks old) were randomly divided into six groups, each group of 5 mice:normal control, model, and low dosage of SZAST, medium dosage of SZAST, high dosage of SZAST and quetiapine(QTP)-treated group. The control group were given mixed cuprizone (CPZ, a copper chelator, 0.2%, w/w) rodent chow for six successive weeks to induce demyelination. During the last two weeks, mice were given an oral gavage of saline, or SZAST of three different doses (a low dose of 5.5 g·kg^-1·d^-1, a medium dose of 11.0 g·kg^-1·d^-1, or a high dose of 16.5 g·kg^-1·d^-1), or QTP, respectively. Then, the mice were anesthetized and their cecum contents removed. Illumina MiSeq was used as the sequencing platform to analyze and evaluate the number of operational taxonomic units (OTUs), richness and diversity indexes, diversity of alpha and beta, differential phylum and genus of intestinal flora in V3-V4 zone of 16S rRNA. Results SZAST could regulate the number of OTUs, richness and diversity indexes of intestinal flora, imbalance of alpha and beta diversity of demyelinated model mice.Compared to the control group, Lachnospiraceae family were decreased in the model group (p<0.05), and SZAST could increase the diversity of Lachnospiraceae family. Compared to the control group, Beta-proteobacteria, Burkholderiales, Alcaligenaceae, and Bacteroidales were increased in the model group (p<0.05), but SZAST could decrease these intestinal flora. Conclusion SZAST had a therapeutic effect on the abnormal flora diversity of demyelinated model mice. It revealed the relevant pathological mechanism of mental schizophrenia caused by white matter damage through 16S rRNA technology.

Key words: schizophrenia, Shi-Zhen-An-Shen-Tang, cuprizone, gut microbiota, high-throughput sequencing, 16S rRNA technique, operational taxonomic units(OTUs)

中图分类号:

马超, 贺毅, 孙作厘, 刘鑫垚, 冯正田, 陈沛, 宁艳哲, 朱虹, 尹冬青, 贾竑晓. 基于16S rRNA技术分析石珍安神汤对脱髓鞘小鼠肠道菌群多样性的影响[J]. 首都医科大学学报, 2020, 41(1): 64-70.

Ma Chao, He Yi, Sun Zuoli, Liu Xinyao, Feng Zhengtian, Chen Pei, Ning Yanzhe, Zhu Hong, Yin Dongqing, Jia Hongxiao. Analysis of effect of Shi-Zhen-An-Shen-Tang on intestinal microflora diversity of mice exposed to cuprizone based on 16S rRNA technique[J]. Journal of Capital Medical University, 2020, 41(1): 64-70.

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基于16S rRNA技术分析石珍安神汤对脱髓鞘小鼠肠道菌群多样性的影响

Analysis of effect of Shi-Zhen-An-Shen-Tang on intestinal microflora diversity of mice exposed to cuprizone based on 16S rRNA technique

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