基于PCR-DGGE分析抗性淀粉对高脂饮食HFA小鼠肠道菌群的影响

PCR-DGGE Based Analysis of the Impact of Resistant Starch on Gut Microbiota in HFA Mice

本文研究了香蕉抗性淀粉对高脂饮食菌群人源化小鼠(HFA)肠道菌群的调控效果。将30只肠道菌群人源化昆明小鼠随机分为3组(10只/组),分别饲喂普通饮食、高脂饮食和高脂饮食+抗性淀粉。提取第0周和第8周新鲜粪便DNA,比较用16S r DNAV6-V8区段特异性引物扩增产物,以PCR-DGGE技术手段检测抗性淀粉组、高脂饮食组和普通饮食组肠道菌群差异。第0周,三组小鼠菌群总菌丰富度,多样性指数和均匀度无显著差异(p>0.05),小鼠和粪便供者之间菌群相似性为36%;三组小鼠肠道菌群相似性达到60-93%,PCA分析显示三组小鼠的数据点都聚在一起,说明在干预前无明显差异,适用于分析抗性淀粉对肠道菌群的影响;膳食干涉第8周后小鼠肠道菌群的多样性指数和丰富度数值,抗性淀粉组都极显著低于高脂组和普通饲料组(p<0.01),PCA分析也显示三组小鼠肠道菌群各组的数据点都分布在不同区域,说明抗性淀粉对高脂饮食HFA小鼠肠道菌群结构产生显著影响。

The effects of banana resistant starch on the alteration of gut microbiota in Human Flora-Associated(HFA) mouse model fed on a high fat diet, were investigated. Thirty HFA mice were randomly assigned into three groups(n = 10/group), which were fed a normal diet, high fat diet, and high fat diet + resistant starch, respectively. DNA was extracted from the fresh feces of all mice, which were collected prior to the beginning of the experiment(0th week) and after eight weeks(8th week). The primer-specific amplification products using the V6-V8 region of 16 S r DNA were compared and differences in the profile of gut microbiota among the resistant starch, high fat diet, and normal diet groups were tested using polymerase chain reaction–denaturing gradient gel electrophoresis(PCR-DGGE). Richness, diversity index, and evenness value of gut microbiota were not significantly different between the three groups at 0th week(p > 0.05). Flora similarity between the mice and the human feces donor was 36%, but the flora similarity between the three mice groups ranged from 60% to 93%. Principal component analysis(PCA) showed that the data points of all three groups were clustered together, indicating no difference between the three groups before intervention and that the HFA mouse model was suitable for the investigation of resistant starch on gut microbiota. Richness and diversity index values of gut microbiota between the resistant starch group and the other two groups 8 weeks after the dietary intervention were significantly different(p < 0.01). PCA of the mouse intestinal bacterial community showed that data points were distributed within different zones after the dietary interventions. In conclusion, resistant starch has a huge influence on the gut bacterial community structure of HFA mice fed on a high fat diet.