七味白术散对菌群失调腹泻小鼠肠道细菌多样性的影响

Effects of Qiweibaizhu powder on the intestinal bacterial diversity in dysbacteriotic diarrhea mice

摘要:

【背景】肠道微生物与人体多项生理功能密切相关，我们课题组前期研究表明七味白术散对菌群失调腹泻有较好的疗效。【目的】探讨与七味白术散疗效相关肠道微生物，比较传统中药饮片与超微中药饮片的疗效，进一步明确七味白术散治疗菌群失调腹泻的机理，为临床应用提供科学依据。【方法】应用抗生素建立菌群失调小鼠腹泻模型，分别灌胃给药七味白术散传统汤剂和超微50%量七味白术散汤剂。治疗结束后，提取肠道内容物微生物宏基因组DNA，建立16S rRNA基因文库进行测序。【结果】肠道微生物中的群落由乳酸菌(Lactobacillus spp.)、屎肠球菌(Enterococcus feacium)、梭状芽孢杆菌(Clostridium spp.)、黏液真杆菌(Blautia producta)、毛螺旋菌(Anaerostipes spp.)、腐生性葡萄球菌(Staphylococcus saprophyticus)和不能培养的细菌组成，其中乳酸菌为优势菌，占全部细菌DNA克隆数的61.90%。经抗生素造模后Lactobacillus spp.数量明显减少，Enterococcus feacium成为优势菌种，Clostridium spp.、Blautia product、Anaerostipes spp.和Staphylococcus saprophyticus等在数量上开始有增长的趋势，而治疗结束后，传统七味白术散治疗组和超微50%量七味白术散治疗组的Lactobacillus spp.比例均有所恢复，且超微50%量七味白术散治疗组的乳酸菌比例与正常组最接近；通过建立聚类树和计算各组中各菌群的多样性(H)、丰富度(S)、均匀度(E)及优势度指数(D)可知，超微50%量七味白术散治疗组可与正常组聚为一类，且超微50%量七味白术散治疗组各项指数与正常组最为接近，说明超微50%量七味白术散治疗组肠道微生物的基因文库及多样性与正常组最接近，超微50%量七味白术散汤剂的治疗效果优于七味白术散传统汤剂。【结论】应用16S rRNA基因克隆文库技术进一步明确了正常小鼠肠道中细菌群落的组成，以及七味白术散对菌群失调腹泻 小鼠肠道细菌群落的恢复效果，超微50%量七味白术散汤剂的治疗效果优于七味白术散传统 汤剂。

Abstract:

[Background] Intestinal microbiota is related to several physiological functions of human body. A large number of studies have shown that Qiweibaizhu powder has effects on diarrhea mice. [Objective] Compare with traditional Chinese medicine and ultra-shattered medicine curative effect. To explore the correlation efficacy with Qiweibaizhu powder to provide scientific evidence for the clinical application. [Methods] Dysbacteriotic diarrheal mice model were constructed via administration of antibiotics and then treated with traditional Qiweibaizhu powder and 50% amount of ultra-shattered Qiweibaizhu powder. After the treatment, 16S rRNA gene of microorganisms in intestinal contents of different groups was analyzed by PCR cloning, and corresponding gene library was constructed. [Results] Lactobacillus spp., Enterococcus feacium, Clostridium spp., Blautia producta, Anaerostipes spp., Staphylococcus saprophyticus and uncultured bacterium were in the normal intestinal microbial flora, and Lactobacillus spp. was the dominant bacteria in the predominant intestinal microbial flora, accounting for 61.90% of the total bacterial DNA clone number. Compared with the control group, the proportion of Lactobacillus spp. in the model group was significantly decreased and opportunistic pathogen was increased. After intrgastric administration, the proportion of Lactobacillus spp. in the traditional Qiweibaizhu powder treatment group and 50% amount of ultra-shattered Qiweibaizhu powder treatment group was recovered, which the proportion of Lactobacillus spp. in 50% amount of ultra-shattered Qiweibaizhu powder treatment group was close to the normal group. Through establishing a clustering tree and calculating the diversity (H), species richness (S) and dominance index (D) of each category, we knew that each index of 50% amount of ultra-shattered Qiweibaizhu powder treatment group were approximately to the control group. In conclusion, the intestine microbial diversity of 50% amount of ultra-shattered Qiweibaizhu powder treatment group was most close to the normal group. [Conclusion] The 16S rRNA gene clone library technique was used to further clarify the composition of bacterial community in normal mice and the recovery of intestinal bacterial community in Qiweibaizhu powder, and the effect of 50% amount of ultra-shattered Qiweibaizhu powder treatment was better than the traditional Qiweibaizhu powder treatment.