基于16S rDNA测序分析腹腔注射苦参碱的昆明小鼠肠道菌群结构

Influence of Matrine on Intestinal Flora of Kunming Mice through Intraperitoneal Injection Based on 16S rDNA Sequencing

旨在通过16S rDNA测序技术分析腹腔注射苦参碱的昆明小鼠肠道菌群的结构。本研究将20只昆明小鼠随机分为2组,分别是苦参碱组(MT组)和阴性对照组(NC组),连续腹腔给药5 d,每天给药2次,收集各组粪便和各肠段组织,进行β多样性、Lefse及Metastats分析,qPCR检测差异菌种在各肠段的mRNA表达量,通过KEGG分析肠道菌群变化导致的代谢途径差异。稀释曲线结果显示,所测样本数据足以反映样品中物种多样性;β多样性分析结果显示,苦参碱可以调节肠道菌群的结构,Lefse及Metastats分析结果均显示,苦参碱显著增加了拟杆菌门(Bacteroidetes)、拟杆菌目(Bacteroidales)、Muribaculaceae、益生菌嗜酸乳杆菌(Lactobacillus acidophilus)的丰度,而显著减少了厚壁菌门(Firmicutes)、瘤胃菌科(Ruminococcaceae)和脱硫弧菌属(Desulfovibrio)的丰度。与Lefse及Metastats分析结果一致,qPCR结果显示苦参碱组小鼠粪便中嗜酸乳杆菌含量增加。同时,苦参碱可以增强嗜酸乳杆菌在各肠段的定植。通过KEGG分析发现,NC与MT组之间在聚糖的生物合成与代谢、运输与分解代谢等代谢途径存在显著差异。本研究结果表明,腹腔注射苦参碱可以显著改变昆明小鼠肠道菌群的结构,增加有益菌嗜酸乳杆菌在肠道中的定植,并造成了聚糖生物合成与代谢、运输与分解代谢等代谢途径的差异,为进一步揭示苦参碱发挥药效作用的机理奠定了基础。

The study aimed to analyse the structure of the intestinal flora of Kunming mice injected with matrine by 16S rDNA sequencing.Twenty Kunming mice were randomly divided into 2 groups named as matrine group(MT group)and negative control group(NC group).The intraperitoneal administration was administered twice a day for 5 days,the feces and intestinal tissues of each group were collected forβdiversity,Lefse and Metastats analysis.qPCR was used to detect the mRNA expression of different strains in each intestinal segment,and KEGG was used to analyze the differences in metabolic pathways caused by the changes in intestinal flora.The rarefaction curve results showed that the measured sample data was sufficient to reflect the species diversity in the samples.The results ofβdiversity analysis showed that matrine regulated the structure of the intestinal flora.Lefse and Metastats analysis results showed that matrine significantly increased the abundance of Bacteroidetes,Bacteroidales,Muribaculaceae and probiotic Lactobacillus acidophilus,and significantly reduced the abundance of Firmicutes,Ruminococcaceae and Desulfovibrio.The qPCR results showed that the expression of Lactobacillus acidophilus in the feces of mice in the matrine group was increased,which was consistent with the results of Lefse and Metastats analysis.It was also found that matrine enhanced the colonization of Lactobacillus acidophilus in various segments of the intestine.KEGG analysis found that there were significant differences in metabolic pathways such as glycan biosynthesis and metabolism,transport and catabolism between NC and MT groups.The study results showed that matrine significantly changed the structure of the intestinal flora in mice,increase the colonization of probiotic Lactobacillus acidophilus in the intestine and caused the differences in metabolic pathways such as glycan biosynthesis and metabolism,transport and catabolism.These results have accumulated data for further study of the mechanism of matrine exerting its pharmacological effects.