基于高通量测序分析多巴胺受体DRD5缺失小鼠肠道菌群的变化

Analysis on changes of gut microbiota in DRD5 knockout mice based on high-throughput sequencing

目的探讨多巴胺受体DRD5缺失小鼠肠道菌群结构的变化和多样性。方法通过基因敲除技术构建DRD5敲除小鼠模型,根据小鼠基因型分为两组:野生型小鼠作为对照组,DRD5敲除小鼠作为敲除组。8周龄时,收集5只对照组小鼠和5只敲除组小鼠的粪便标本,利用Illumina MiSeq测序平台对细菌的16S rRNA进行高通量测序,对测序结果进行生物信息学分析。结果敲除组与对照组小鼠的肠道菌群结构存在差异。在门水平上,敲除组与对照组优势菌门拟杆菌门和厚壁菌门的相对丰度比较,差异无统计学意义(P>0.05);变形菌门和蓝细菌门两组间比较,差异有统计学意义(P<0.05)。在属水平上,敲除组与对照组比较,罗姆布茨菌属、真杆菌属、脱硫弧菌属、ⅩⅢ_UCG-001属、胃嗜气科菌属、Rs-E47_termite属、双歧杆菌属、ASF356属、布劳特菌属、毛螺菌属的相对丰度差异有统计学意义(P<0.05)。进一步对菌群进行分析,敲除组的罗姆布茨菌属、真杆菌、脱硫弧菌属、ⅩⅢ_UCG-001属、Rs-E47_termite属、双歧杆菌属、ASF356属和毛螺菌科NK4A136组的相对丰度均低于对照组(P<0.05),胃嗜气科菌属和布劳特菌属的相对丰度均高于对照组(P<0.05)。结论两组小鼠的菌群构成及构成比皆存在差异,DRD5缺失后会引起小鼠肠道微生态失调。

Objective To investigate the changes in the structure and diversity of intestinal flora in dopamine receptor DRD5-deficient mice.Methods The DRD5 knockout mouse model was constructed by gene knockout technology and the mice were divided into two groups according to the genotype:wild type mice as the control group and DRD5 knockout mice as the knockout group.At 8 weeks of age,fecal samples of 5 control group mice and 5 knockout group mice were collected,and 16S rRNA of bacteria was sequenced by Illumina MiSeq sequencing platform,and the sequencing results were analyzed bioinformatically.Results There was difference in gut microbiota between control and knockout group.At the phylum level,there was no significant difference in the relative abundance of Bacteroidetes and Firmicutes(P>0.05).There was significant difference in the structure of Proteobacteria and Cyanobacteria between the 2 groups(P<0.05).At the genus level,there were significant differences in the relative abundance of Romboutsia,Eubacterium,Desulfovibrio,FamilyⅩⅢ_UCG-001,Gastranaerophilales,Rs-E47_termite,Bifidobacterium,ASF356,Blautia and Lachnospiraceae NK4A136 between the 2 groups(P<0.05).Further analysis of gut microbiota showed that the relative abundance of Romboutsia,Eubacterium,Desulfovibrio,FamilyⅩⅢ_UCG-001,Rs-E47_termite,Bifidobacterium,ASF356 and Lachnospiraceae NK4A136 in the knockout group were lower than those in the conrol group(P<0.05),but the relative abundance of Gastranaerophilales and Blautia in the knockout group were higher than those in the control group(P<0.05).Conclusion There were differences in the composition and composition ratio between the two groups of mice,and DRD5 deletion caused intestinal microecological disorders in mice.