基于16S rRNA测序技术研究痛泻要方调节腹泻型肠易激综合征模型大鼠肠道菌群的作用机制

Effects of Tong-Xie-Yao-Fang on Intestinal Flora of Rats with Diarrhea Irritable Bowel Syndrome and Its Mechanism Interpretation Based on 16S rRNA Sequencing

目的探讨痛泻要方是否可以通过调节肠道菌群而对腹泻性肠易激综合征大鼠发挥治疗作用。方法将24只SD大鼠随机分为4组:空白组、模型组、痛泻要方组和阳性药组。采用番泻叶灌胃加束缚刺激的方法制备大鼠腹泻型肠易激综合征模型。建模成功后连续给药1周。对大鼠进行体质量变化、病理检测、AWR评分等药理学效应指标检测,并对产生腹泻症状的大鼠计数。同时,通过16S rRNA高通量测序,对大鼠的肠道菌群进行测序分析。结果所采用的方法可以成功制备大鼠腹泻型肠易激综合征模型。经痛泻要方治疗后,模型大鼠的腹泻症状得到缓解;痛泻要方组大鼠体质量及AWR评分明显高于模型组(P<0.001)。与模型组相比,痛泻要方组大鼠肠道菌群的丰度、多样性增加;在门水平下分析,痛泻要方组大鼠体内的拟杆菌门丰度增加,变形菌门、螺旋体门和疣微菌门的丰度降低。结论痛泻要方能够通过调节肠道菌群的丰度及组成,对腹泻型肠激综合征大鼠发挥治疗作用。

OBJECTIVE To investigate whether Tong-Xie-Yao-Fang(TXYF)could play a therapeutic role in IBS-D by regulating intestinal flora.METHODS In this paper,twenty-four Sprague Dawley(SD)rats were randomly divided into four groups:control,IBS-D model,TXYF and pinaverium bromide positive groups.IBS-D model was established by given senna decoction intragastrically and chronic restraint stress.Then,these IBS-D model rats were treated with TXYF.Traditional pharmacology indexes,including body weight variety,pathological examination and abdominal withdrawal reflex(AWR)scores were tested.The number of rats that suffered with diarrhea were also counted.Furthermore,fecal microbiota alterations were assayed by 16S rRNA high-throughput sequencing.RESULTS Pharmacology indexes and diarrhea rate of model group indicated that IBS-D model was established successfully.According to the results,diarrhea of IBS-D rats could be controlled by TXYF treatment.Body weight and AWR scores(P<0.001)of TXYF group were found significantly higher than model group.Compared with model group,the results of fecal microbiota showed that the community richness and diversity for TXYF group were improved.Fecal microbiota analyzed in phylum displayed that the abundance of Bacteroidetes was increased,while those of Proteobacteria,Spiroch-aetae and Verrucomicrobia were decreased.CONCLUSION TXYF is proved to be beneficial to IBS-D rats through modifying the abundance of gut microbiota.