基于16S rDNA测序分析中药复方石苦秦散对腹泻小鼠盲肠的影响

Effects of the Traditional Chinese Medicine“Shikuqin”Powder on Caeca in Diarrhea Mice Based on 16S rDNA Sequencing

【目的】探明止泻中药复方石苦秦散对蓖麻油和番泻叶所致腹泻小鼠肠道微生物的影响,为研究石苦秦散止泻作用机制奠定基础。【方法】将70只小鼠分为正常对照组、阳性药物组、蓖麻油模型组、蓖麻油中药预防组、番泻叶模型组、番泻叶中药预防组和番泻叶中药治疗组,按试验设计进行灌胃处理后,观察各组小鼠腹泻情况,针对不同药物腹泻模型采集十二指肠、盲肠观察肠道病理变化,最后采集各组盲肠内容物进行16S rDNA测序。【结果】腹泻指数显示,灌胃1 h番泻叶中药预防组和中药治疗组均极显著(P<0.01)低于番泻叶模型组;蓖麻油模型灌胃2 h与正常对照组差异极显著。肠道病理变化主要为制造腹泻模型后黏膜下层有少量炎性细胞浸润,除正常对照组外其他组肌层和浆膜变薄。盲肠Alpha测序显示,蓖麻油造模后,中药预防组能显著提高盲肠菌群丰度和多样性;Beta多样性PCoA分析表明,蓖麻油模型组与其他组样本明显分开;物种分类显示,2种药物腹泻模型处理后中药预防组和治疗组均不同程度提高拟杆菌门(Bacteroidetes)、软壁菌门(Tenericutes)丰度,降低变形菌门(Proteobacteria)丰度;属水平在正常范围内,降低乳杆菌(Lactobacillus)和大肠杆菌(Escherichia)丰度,提高有益菌Mucispirillum和拟普雷沃菌属(Alloprevotella)丰度。【结论】蓖麻油和番泻叶小鼠腹泻模型使用中药复方石苦秦散预防和治疗后,可明显改善肠道微生物丰度,特别是炎症反应相关的微生物,说明石苦秦散可通过调整肠道炎症修复发挥止泻作用。

【Objective】The effects of the traditional Chinese medicine“Shikuqin”powder on intestinal microorganisms of diarrhea mice caused by castor oil and folium sennae were studied,which provided a basis for studying the antidiarrheal mechanism of Shikuqin powder.【Method】70 mice were divided into group for normal control,group for positive-drug,group for castor oil model,group for prevention of castor oil by traditional Chinese medicine,group for folium sennae model,group for prevention of folium sennae by traditional Chinese medicine and group for curing folium sennae by traditional Chinese medicine.After intragastric administration according the experimental design,the diarrhoea situations of 70 mice were observed.The pathological changes in duodenums and caeca of mice were collected according to different models of diarrhoea caused by different drugs,and then the matters in caecum were collected for 16S rDNA sequencing.【Result】The diarrhea index after intragastric administration for one hour,the group for prevention of folium sennae by traditional Chinese medicine and group for curing folium sennae by traditional Chinese medicine were both extremely significantly(P<0.01)lower than group for folium sennae model.After intragastric administration for 2 hours,the mice from group for castor oil model were extremely significantly different from mice from group for normal control.The pathological changes of intestinal tract mainly showed that there was a little inflammatory cell infiltration in the submucosa after the diarrhea model was made,and the muscle layer and serous membrane became thinner in other groups except the normal control group.Cecal Alpha sequencing showed that the abundance and diversity of cecal flora could be significantly increased in the group for prevention by traditional Chinese medicine after castor oil modeling.PCoA analysis of Beta diversity showed that the castor oil model group was significantly separated from other group.The species classification showed that after the treatment of the two drug diarrhea models,the abundance of Bacteroidetes and Tenericutes were increased in the prevention or treatment group,while the abundance of Proteobacteria was decreased in different degrees.Genus levels stayed in the normal range,as the abundance of Lactobacillus and Escherichia decreased and the abundance of beneficial bacteria Mucispirillum and Alloprevotella increased.【Conclusion】For the mouse model of diarrhea caused by castor oil and senna leaf,the abundance of intestinal microorganisms can be significantly improved after the prevention and treatment with compound Shikunqin powder,especially the microorganisms related to inflammatory response,indicating that Shikuqin powder may play an antidiarrheal effect by adjusting the repair of intestinal inflammation.