基于16S rDNA测序分析地榆七柏汤对溃疡性结肠炎大鼠肠道菌群的影响

Effect of Diyuqibai Decoction on intestinal microflora in ulcerative colitis rats based on 16S rDNA sequencing

目的观察地榆七柏汤对湿热型溃疡性结肠炎大鼠肠道菌群的影响,探讨其治疗溃疡性结肠炎的作用机制。方法取84只Wistar大鼠,随机选择9只作为空白组,其余大鼠采用高糖高脂饮食+葡聚糖硫酸钠法建立湿热型溃疡性结肠炎模型。将造模成功大鼠随机分为模型组、柳氮磺胺吡啶组及地榆七柏汤低、中、高剂量组,每组14只。柳氮磺胺吡啶组给予柳氮磺胺吡啶混悬液300 mg/kg灌肠,地榆七柏汤低、中、高剂量组分别给予含生药0.5 g/mL、1 g/mL、3 g/mL的地榆七柏汤保留灌肠,模型组和空白组给予等量生理盐水灌肠,均1次/d,连续干预14 d。观察大鼠一般状况,进行疾病活动度指数(DAI)评分,HE染色观察结肠组织病理形态,采用16S rDNA测序技术分析粪便菌群物种组成情况。结果与模型组比较,各药物组大鼠腹泻、便血等症状明显好转,DAI评分明显降低(P均<0.05),结肠组织病理损伤明显减轻。菌群构成方面,模型组大鼠毛螺菌属(Lachnospiraceae)、Mucispirillum属丰度均明显低于空白组(P均<0.05),各药物组均明显高于模型组(P均<0.05);模型组大鼠肠杆菌科(Enterobacteriaceae)丰度明显高于空白组(P<0.05),各药物组均明显低于模型组(P均<0.05)。结论地榆七柏汤可明显减轻湿热型溃疡性结肠炎大鼠疾病活动度和肠黏膜炎性损伤,其机制可能与调节肠道菌群、提升有益菌丰度、降低致病菌丰度相关。

Objective It is to observe the effect of Diyuqibai Decoction on intestinal flora of rats with ulcerative colitis(UC)of damp-heat type and explore its mechanism in the treatment of UC.Methods Eighty-four Wistar rats were taken,in which 9 rats were randomly selected as the blank group,and the rest of the rats were used to establish the models of damp-heat UC by the method of high-sugar and high-fat diet plus dextran sodium sulfate.The successfully modeled rats were randomly divided into model group,sulfasalazine(SASP)group and low,medium and high dose groups of Diyuqibai Decoction,with 14 rats in each group.The SASP group was given SASP suspension 300 mg/kg by enema,the low,medium and high dose groups of Diyuqibai Decoction were given medicated serum containing 0.5 g/mL,1 g/mL and 3 g/mL of Diyuqibai Decoction by retained enema,respectively,while the model group and blank group were given equal amounts of normal saline by enema,all once daily,continuously treated for 14 days.The general condition of the rats in each group was observed and scored by disease activity index(DAI),the histopathological changes of their colon tissue were observed by HE staining,the fecal flora species composition was analyzed by 16S rDNA sequencing.Results Compared with the model group,the symptoms such as diarrhea and blood stool of rats in each treatment group were significantly improved,their DAI score were significantly reduced(all P<0.05),and the histopathological damage of colon tissue was significantly alleviated.As for the composition of bacterial flora,the abundance of Lachnospiraceae and Mucispirillum in rats of the model group were significantly lower than those in the blank group(all P<0.05),and the abundances in each treatment group were significantly higher than those in the model group(all P<0.05);the abundance of Enterobacteriaceae in the model group was significantly higher than that in the blank group(P<0.05),and the abundance in each treatment group was significantly higher than that in the mdel group(P<0.05).Conclusion Diyuqibai Decoction can significantly reduce disease activity and improve intestinal mucosal inflammatory injury in rats with damp-heat ulcerative colitis.Its mechanism may be related to regulate intestinal flora,increase the abundance of beneficial bacteria and reduce the abundance of pathogenic bacteria.