基于16S rRNA基因测序技术分析急性肝内胆汁淤积大鼠的肠道菌群
Analysis of intestinal flora in rats with acute intrahepatic cholestasis based on 16S rRNA gene sequencing
黄一曼 1孙凤霞 2李晓玲2
作者信息
- 1. 北京中医药大学,北京 100029;首都医科大学附属北京中医医院肝病科
- 2. 首都医科大学附属北京中医医院肝病科
- 折叠
摘要
目的 探讨 ANIT诱导的大鼠急性肝内胆汁淤积(acute intrahepatic cholestasis,AIC)与肠道菌群的关系.方法 以 SPF级的 SD大鼠为实验对象,随机分为正常对照组、模型组,每组 24 只,依据取材时间不同又将各组随机分为 24 h、48 h、72 h 3 个亚组,每组 8 只.模型组通过一次性予 2%ANIT按 100 mg·kg-1·d-1 灌胃造模诱发AIC模型,正常对照组予等容积色拉油灌胃.通过高通量测序法对肠道粪便细菌 16S rRNA的 V3～V4 区进行基因测序及生物信息学分析.结果 AIC造模后,随着时间增加,α 多样性指数逐步升高,造模 72 h组相比造模 24 h组,Shannon指数有显著性差异;β多样性发现,利用基于加权Unifrac距离PcoA分析或 NMDS分析均显示:正常对照组、造模 24 h组和造模 48 h组的肠道菌群显示比较明显的聚集效应;在门水平上,厚壁菌门(Firmi-cutes)丰度在造模 48 h后显著下降,变形菌门(Proteobacteria)的丰度显著上升,拟杆菌门(Bacteroidetes)的丰度显著上升;LEfSe 对组间差异显著的物种分析结果显示,LDA值大于 4 的微生物类群有 35 个.结论 模型组干预后大鼠肠道菌群结构和多样性均发生显著变化.
Abstract
Objective To investigate the correlation between rat gut microbiota and ANIT induced acute intrahepatic cholestatic(AIC).Methods SPF SD rats were randomly divided into normal control group and model group.Accord-ing to the time of sampling,each group was divided into 24 h,48 h,and 72 h subgroups,with 8 rats in each subgroup.Rats of model group were given a single dose of 2%ANIT(100 mg·kg-1·d-1)by gavage to establish the model.Changes in the gut microbiota composition were analyzed using 16S rRNA sequencing.Results After AIC modeling,α diversity index gradually increased with the increase of time.The Shannon index had significant difference after 72 h modeling compared with 24 h modeling,β diversity found that PcoA analysis or NMDS analysis based on weighted Unif-rac distance showed that:the intestinal flora in the control group,24 h and 48 h after modeling showed obvious aggrega-tion effect.At the phyla level,the abundance of Firmicutes decreased significantly 48 h after modeling,the abundance of proteobacteria increased significantly,and the abundance of Bacteroides increased significantly.The results of species analysis with significant differences between groups by LEfSe showed that there were 35 microbial groups with LDA val-ues greater than 4.Conclusion The structure and diversities of gut microbiota of model group rats were significantly changed compared with normal control group.
关键词
急性肝内胆汁淤积/大鼠/16S/rRNA基因测序技术/肠道菌群Key words
Acute intrahepatic cholestasis/Rat/16S rRNA sequencing/Gut microbiota引用本文复制引用
基金项目
首都医科大学附属北京中医医院中医药学科建设项目()
首都医科大学临床专科学院(系)培养基金开放课题(CCMU2022ZKYXZ008)
首都卫生发展科研专项项目(首发2018-2-2173)
北京市属医院科研培育计划项目(PZ2018023)
出版年
2024
NETL
NSTL
万方数据