抗生素和高脂饮食对大鼠肠道乳杆菌多样性的影响

Effect of antibiotics or high-fat diet on diversity of intestinal Lactobacillus spp. in rat

摘要:

【目的】对正常、高脂、抗生素处理大鼠肠道内乳杆菌进行定性和定量分析，比较不同处理组大鼠肠道乳杆菌的多样性。【方法】应用纯培养和非培养技术(16S rRNA基因序列分析、变性梯度凝胶电泳、实时荧光定量PCR)对大鼠肠道乳杆菌进行分离鉴定和多样性分析。【结果】16S rRNA基因序列同源性分析结果显示，正常组大鼠肠道内分离出的乳杆菌包括约氏乳杆菌(Lactobacillus johnsonii)、鼠乳杆菌(Lactobacillus murinus)、嗜酸乳杆菌(Lactobacillus acidophilus)、罗伊氏乳杆菌(Lactobacillus reuteri)、植物乳杆菌(Lactobacillus plantarum)、肠道乳杆菌(Lactobacillus intestinals)、动物乳杆菌(Lactobacillus animalis)和阴道乳杆菌(Lactobacillus vaginalis)；但L. animalis在高脂处理组大鼠肠道内未分离到，L. intestinals和L. vaginalis在抗生素处理组大鼠中未分离到。DGGE结果显示3个组别大鼠肠道中乳杆菌构成差异明显，同一组内样品间相似性较高；相较于正常组和高脂组，抗生素组的丰度较差；且正常组大鼠肠道内乳杆菌的多样性高于高脂组和抗生素组。q-PCR结果显示正常组大鼠肠道乳杆菌的数量明显高于高脂组和抗生素组，高脂组的数量也明显高于抗生素组，且3个组别之间存在显著差异(P<0.01)。【结论】高脂饮食及抗生素的使用会减少肠道内乳杆菌多样性。

Abstract:

[Objective] This study aimed to investigate the diversity of intestinal Lactobacillus in normal diet rats (NDR), high-fat diet rats (HFDR) and antibiotic diet rats (ADR). [Methods] Culture-dependent and culture-independent methods (16S rRNA gene sequencing, DGGE and q-PCR) were applied to isolation, identification and diversity analysis of intestinal Lactobacillus in above three groups. [Results] According to the 16S rRNA gene sequencing analysis, L. johnsonii, L. murinus, L. acidophilus, L. reuteri, L. plantarum, L. intestinals, L. animalis and L. vaginalis were isolated from the intestinal of NDR. Whereas L. animalis were not able to find in HFDR, L. intestinals and L. vaginalis were not isolated from ADR. The DGGE results showed that there was an obvious contrast in the composition of intestinal Lactobacillus among 3 groups, and a high similarity in the same group as well. Comparing the richness of Lactobacillus in three groups, ADR was the lowest one. Moreover, the intestinal Lactobacillus of NDR owed the highest diversity than the others. The q-PCR data indicated that the population of Lactobacillus in NDR was higher than that in other two groups, there was a significantly different (P<0.01) among three groups. [Conclusion] The diversity of the intestinal Lactobacillus can be reduced with the interference of high-fat diet or antibiotic.