用DGGE技术分析植物乳杆菌对小鼠肠道菌群失调的影响
Effect of Lactobacillus plantarum on disordered intestinal flora of mice introduced by antibiotics using PCR-DGGE technology
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摘要: 采用PCR-DGGE方法研究植物乳杆菌对小鼠肠道失调菌群微生态的影响。选用昆明雌性小鼠建立小鼠抗生素相关性菌群失调模型,灌服植物乳杆菌菌液(109CFU/mL)5d,分离提取小鼠粪便中微生物群落的DNA,通过PCR-DGGE技术分析植物乳杆菌调节过程微生物群落结构与多样性的影响。PCR-DGGE图谱显示,随着植物乳杆菌饲喂时间的延长,小鼠肠道菌群丰富度呈现逐渐增加的演替过程。植物乳杆菌灌胃第4d小鼠肠道菌群的丰富度指数(18)与多样性指数(2.75)与正常小鼠基本一致,DGGE图谱的测序结果表明,植物乳杆菌调节小鼠肠道菌群失调主要表现在两个方面:选择性地增加有益菌如Bacteroides stercoris、Bacteroides uniformis strain等的量;抑制致病菌如Clostridium clostridioforme、Pseudomonas stutzeri等的生长。说明植物乳杆菌对头孢地尼引起的肠道菌群失调具有调整作用;建立PCR-DGGE法监测肠道菌群华体会(中国)变化的分析方法,为进一步研究益生菌对肠道菌群的作用机制奠定基础。Abstract: This article’s aim was to adopt PCR -DGGE method to study the effect of feeding Lactobacillus plantarum to antibiotic intestinal flora in mice on the gut microflora. Female KM mice were continuously fed with cefdinir solution for five days to cause antibiotic-associated intestinal flora in them, and then treated by oral gavage of Lactobacillus plantarum (10 9 CFU/mL) for another 5 days. Feces were abstracted and 16S rDNA-based PCR-DGGE method was used to analyze the influence of Lactobacillus plantarum adjustment process on the microbial community structure and diversity. The main preponderant strips were rubber cutting cloned and sequenced. PCR-DGGE profiles showed that with the extension of the Lactobacillus plantarum feeding time, the richness of the mouse intestinal flora presented a gradual increase in the successionprocess. The richness index (18) and diversity index (2.75) of the intestinal flora after the mice were gavaged using Lactobacillus plantarum for fourth day were the same as the normal ones. DGGE profiles, sequencing results showed that plant lactobacillus control mice intestinal dysbacteriosis mainly manifested in two aspects:selective increase beneficial bacteria such as Bacteroides stercoris, Bacteroides uniformis strain of quantity. Inhibition of pathogenic bacteria such as Clostridium clostridioforme, Pseudomonas stutzeri growth, etc. Results showed that the Lactobacillus plantarum had adjustment effect on intestinal flora disorder caused by cefdinir. Establishment of PCR-DGGE method analysis methods were in order to monitor the dynamic changes of intestinal flora, and to lay the foundation for further study of probiotics on the mechanism of action of the intestinal flora.