基于16S rRNA基因测序比较分析不同抗生素干预对小鼠肠道菌群动态变化的影响

Use of 16S rRNA high-throughput sequencing for comparative analysis of dynamic changes of gut microbiome in mice after intervention with different antibiotics

目的探讨不同种类抗生素对小鼠肠道菌群多样性和组成动态变化的影响。方法采用健康SPF级雌性C57BL/6小鼠20只随机分为对照组(CON组)6只、万古霉素组(VAN组)6只和多黏菌素组(PMB组)8只,通过饮用抗生素方式连续处理5周,于3周、5周采集小鼠粪便样本进行16S rRNA基因的V4-V5区测序。结果与CON组相比,VAN组处理3周和5周菌群多样性显著降低,以疣微菌门(阿克曼菌科)、变形菌门(肠杆菌科)及无胆支原体科相对丰度显著增加,拟杆菌门(Muribaculaceae菌科、普雷沃菌科)和毛螺菌科显著减少为特征(P均<0.05)。PMB组处理3周后,菌群多样性显著增加,以普雷沃菌科和毛螺菌科相对丰度显著增加,疣微菌门(阿克曼菌科)和Muribaculaceae菌科显著减少为特征。PMB处理5周后,菌群多样性显著降低,以疣微菌门(阿克曼菌科)相对丰度显著增加,而厚壁菌门(毛螺菌科)显著减少为特征(P均<0.05)。结论用不同种类的抗生素处理不同时间后小鼠肠道菌群组成不同,但多样性均显著降低,且抗生素处理时间越长菌群多样性降低幅度越大,菌群种类越少。

Objective This study was aimed to explore the dynamic changes in diversity and composition of gut microbiota of C57 BL/6 mice treated with different antibiotics for different durations. Methods Twenty SPF mice were randomly divided into control group(CON group), vancomycin group(VAN group) and polymyxin group(PMB group), respectively. Antibiotics were added into the drinking water, which were continuously used for 5 weeks. Fresh fecal samples from mice were collected once a week. Altogether, 40 samples were collected and their total DNAs were extracted. Sequencing was performed on bacterial 16 S rRNA V4-V5 region. Results The administration of different antibiotics did not affect the growth of mice. Compared with the controls, mice who were treated by vancomycin 3 and 5 weeks had significantly increased abundances in phyla of Verrucomicrobia and Proteobacteria as well as families of Akkermansiaceae, Enterobacteriaceae and Acholeplasmataceae, while their Bacteroidetes phylum and families of Muribaculaceae, Prevotellaceae, Lachnospiraceae were significantly decreased(all P<0.05). For mice who were treated by polymyxin for 3 weeks, the relative abundances in families of Prevotellaceae, and Lachnospiraceae were significantly increased, while Verrucomicrobia phylum, families of Muribaculaceae and Akkermansiaceae were significantly decreased. For mice who were treated by polymyxin for 5 weeks, the relative abundances in Verrucomicrobia phylum and Akkermansiaceae family were significantly increased, while those in phylum Firmicutes and family Lachnospiraceae were significantly decreased(all P<0.05). Conclusion Our results indicated that the richness and diversity of gut microbiota in mice were dramatically decreased after the treatment by vancomycin and polymyxin, the longer the use of these antibiotics, the higher the decrease of microbiome diversity. The finding of this study will provide insights into the community and diversity of gut microbiome in mice after treatment with different antibiotics and will help understand the effect of the long-term use of antibiotics on alterations in gut microbiome of patients.