理中汤对抗生素相关性腹泻模型构建中肠道菌群变化的影响  

Influence of Lizhong Tang Intestinal Flora Changes in Its Treatment to AAD Model

作　　者：舒青龙[1] 王萍[1] 封勇[1] 冯洁[1] 

机构地区：[1]江西中医药大学,南昌330004

出　　处：《中国实验方剂学杂志》2015年第21期82-87,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81473455;31560038);江西省自然科学基金项目(20122BAB205073);江西省教育厅项目(GJJ13598)

摘　　要：目的：观察抗生素相关性腹泻（AAD）模型构建过程中肠道菌群结构变化及理中汤对AAD模型肠道菌群结构的影响。方法：先选取81只雄性BALB/c小鼠进行预实验,用高、中、低剂量（5,1,0.5 g·kg-1）氨苄青霉素构建肠道菌群紊乱模型,再灌胃艰难梭菌（1.6×108CFU）构建AAD模型,并用高、中、低剂量（7.5,5.5,3.5 g·kg-1）理中汤进行治疗。按动物表型变化得出最佳造模及治疗条件,再选取20只雄性BALB/c小鼠,随机分为4组（各组n=5）：空白组、肠道菌群紊乱组、AAD组、理中汤治疗组（7.5 g·kg-1）按最优化步骤进行造模及治疗,各组粪便混合为样品XL1,XL2,XL3,XL4,应用16S rRNA高通量测序技术研究其菌群变化。结果：AAD模型构建的表型变化符合AAD临床表现。XL1~XL4 4组样品菌群基于最小操作分类单元（OTUs）的变化为：860,282,385,394个;基于门水平的丰度变化为：变形菌门（6.94%,41.77%,1.99%,2.89%）,厚壁菌门（44.34%,34.89%,36.82%,32.11%）,拟杆菌门（46.6%,22.74%,60.8%,64.49%）;基于属水平的数量变化为：88,58,52,59个,各组间菌群变化明显,（P〈0.05）。结论：AAD模型构建过程中肠道菌群结构变化剧烈,多样性及丰度均下降;理中汤能改变AAD模型肠道菌群结构,增加菌群多样性。Objective： To observe the intestinal flora changes during the construction of antibiotic associated diarrhea（ AAD） model and the effect of Lizhong Tang on intestinal flora changes in AAD model.Method： Eighty-one male BALB / c mice were selected forpreliminary experiment. High,medium and low dose（ 5,1,0. 5 g·kg- 1） ampicillin was used to construct intestinal flora disorder model,then the mice were fed with Clostridium difficile（ 1. 6 × 108 CFU,colony-forming units） to construct the AAD model,and finally were treated with high,medium and low dose（ 7. 5,5. 5,3. 5 g·kg- 1） Lizhong Tang respectively. According to the animal phenotype and optimum treatment conditions during preliminary experiment,20 male BALB / c mice were selected and randomly divided into 4 groups（ n = 5 in each group） ： blank group,intestinal flora disorder group,AAD group,Lizhong Tang treatment group（ 7. 5 g ·kg- 1）. Modeling and treatment were conducted according to optimization steps. Fecal samples in four groups were respectively mixed as XL1,XL2,XL3,XL4,and then were subjected to flora changes analysis using 16 S rRNA high-throughput sequencing technology. Result： The phenotypic changes during AAD model building were consistent with clinical AAD symptom. The intestinal flora changes at OTUs level were 860,282,385,and 394,respectively; the abundance changes at phylum level were as follows： Proteobacteria（ 6. 94%,41. 77%,1. 99%,2. 89%）,Firmicutes（ 44. 34%,34. 89%,36. 82%,32. 11%）,Bacteroidetes（ 46. 6%,22. 74%,60. 8%,64. 49%） respectively; the number changes at genus level were 88,58,52,and 59 respectively. There were significant flora changesin all groups（ P〈 0. 05）.Conclusion： The intestinal flora structures were changed drastically with decreased diversity and abundance during the construction of AAD model; Lizhong Tang could change the structure of intestinal flora in AAD model,and increase the diversity of flora.

关 键 词：抗生素相关性腹泻 理中汤 肠道菌群 微生态 

分 类 号：R285.5[医药卫生—中药学]