SPF级雄性SD大鼠回肠与结肠菌群结构比较分析  

Analysis on Ileum and Colon Microflora of SPF Male SD Rats based on High-throughput Sequencing

作　　者：许琴[1] 倪艳 是文辉[1] 李建瑛[1] 刘江伟[1] 赵红琼[2] 徐新明 XU Qin;NI Yan;SHI Wenhui;LI Jianying;LIU Jiangwei;ZHAO Hongqiong;XU Xinming(General Hospital of Xinjiang Military Districts,Key Laboratory of Special Environmental Medicines of Xinjiang,Urumqi 830000,China;Veterinary Medicine College of Xinjiang Agricultural University,Urumqi 830052,China;Biotechnology Research Institute of Xinjiang Animal Husbandry Sciences Academy,Urumqi 830000,China)

机构地区：[1]新疆军区总医院,新疆特殊环境医学重点实验室,乌鲁木齐830000 [2]新疆农业大学动物医学学院,乌鲁木齐830052 [3]新疆畜牧科学院生物技术研究所,乌鲁木齐830000

出　　处：《实验动物与比较医学》2023年第1期53-60,共8页Laboratory Animal and Comparative Medicine

基　　金：全军实验动物专项科研课题“高原缺氧寒冷应激环境功能性肠病动物模型制作及干预”(SYDW〔2018〕10)。

摘　　要：目的使用高通量测序技术分析SPF级雄性SD大鼠回肠与结肠菌群结构与丰度。方法收集30只SPF级雄性SD大鼠的回肠及结肠腔内容物,提取内容物中细菌总DNA并进行PCR扩增。使用Ilumina NovaSeq测序平台对样本中细菌16S rRNA的V3-V4区进行测序。然后基于有效数据在门、属水平上分析肠道菌群的物种结构和丰度,采用QIIME软件(扩增子)分析工具分析回肠和结肠菌群的多样性及差异,同时应用Tax4Fun程序预测回肠及结肠菌群的优势基因富集通路。结果在门水平上,雄性SD大鼠回肠内微生物优势菌群主要是厚壁菌门(Firmicutes)和放线菌门(Actinobacteria),占比超过98%;结肠内微生物优势菌群主要是厚壁菌门(Firmicutes)和拟杆菌门(Bacteroidetes),占比超过95%。在属水平上,回肠以乳酸杆菌属(Lactobacillus)和另枝菌属(Alistipes)为主要优势菌群,结肠以乳酸杆菌属(Lactobacillus)及罗姆布茨菌属(Romboutsia)为优势菌群。在菌群多样性方面,结肠菌群的α多样性包括丰富度指数(Chao1指数)及多样性指数(香农指数Shannon index)都显著高于回肠(P<0.01),其菌群物种的组成结构差异性比回肠小;回肠菌群结构差异性显著的主要是厚壁菌门(Firmicutes)、罗姆布茨菌属(Romboutsia)和消化链球菌科(Peptostreptococcaceae),而结肠菌群结构差异性显著的是拟杆菌门(Bacteroidetes)、拟杆菌纲(Bacteroidia)和拟杆菌目(Bacteroidales)。在菌群功能方面,回肠菌群优势基因显著富集于类脂物代谢、多酮代谢、膜运输、生物降解等通路,而结肠菌群优势基因显著富集于甘聚糖生物合成代谢、能量代谢、辅助因子和维生素及其他产物的生物合成等通路。结论SPF级雄性SD大鼠回肠与结肠菌群在结构与丰度上存在显著差异,结肠菌群的丰富度及多样性高于回肠。Objective To analyze the microflora structure and abundance of ileum and colon of SPF male SD rats by high-throughput sequencing technique.Methods The ileal and colonic lumen contents of 30 SPF male SD rats were collected,the total bacterial DNA in the contents was extracted and amplified by PCR.The V3-V4 region of bacterial 16S rRNA in the samples was sequenced using Illumina NovaSeq sequencing platform.Then the species structure and abundance of the intestinal flora were analyzed at the phylum and genus level based on validated data.The diversity and differences between ileal and colonic flora were analyzed using the QIIME software(amplicon)analysis tool.The Tax4Fun program was applied to predict the dominant gene enrichment pathways of ileal and colonic flora.Results At the phylum level,the dominant microflora in the ileum of male SD rats were Firmicutes and Actinobacteria,accounting for more than 98%.The dominant colonic microbial community were mainly Firmicutes and Bacteroidetes,accounting for more than 95%.At the genus level,Lactobacillus and Alistipes were the predominant bacteria in the ileum.The colon was dominated by Lactobacillus and Romboutsia.In terms of flora diversity,the alpha diversity of colonic flora including richness index(Chao1 index)and diversity index(Shannon index)were significantly higher than those of ileum(P<0.01),and the structural variability of the composition of its flora species was smaller than that of ileum;the ileal flora with significant structural variability were mainly of the phylum Firmicutes,Romboutsia,Peptostreptococcaceae,while the colonic flora had significant structural differences in Bacteroidales.In terms of flora function,the ileal flora dominant genes were significantly enriched in lipid-like metabolism,polyketide metabolism,membrane transport,biodegradation and other pathways,while the colonic flora dominant genes were significantly enriched in glycan biosynthesis metabolism,energy metabolism,biosynthesis of cofactor and vitamins and other products,and other pat

关 键 词：肠道菌群 菌群丰度 回肠 结肠 SD大鼠 

分 类 号：Q95-33[生物学—动物学] R-332[医药卫生]