基于肠道菌群的灶心土温脾止泻药效及机制研究

Wenpizhixie(温脾止泻)Effect and Mechanism of Zaoxintu(灶心土)in Treating Diarrhea Based on Gut Microbiota

目的:研究灶心土对脾阳虚泄泻模型小鼠的治疗作用。方法:取昆明小鼠60只,按随机数字表法分为正常对照组、模型对照组、灶心土4、8、12 g/kg组和蒙脱石散混悬液1.5 g/kg组。除正常对照组外,各组小鼠采用“饮食失节+苦寒泻下”的方法建立脾阳虚泄泻小鼠模型。第11 d起,灶心土各组分别灌胃相应药物或生理盐水,1次/d,连续给药7 d。观察小鼠给药前后精神状态及体质量的变化,计算腹泻率和稀便率,采用HE染色观察小鼠结肠和胃病理学改变并按相关标准评分。基于16S rRNA基因测序技术测定小鼠肠道菌群分布情况,分析多样性、菌群丰度及构成,寻找差异菌群。结果:与模型对照组比较,蒙脱石散1.5 g/kg组和灶心土12 g/kg组体质量显著增加(P<0.01)。腹泻率和稀便率显著降低(P<0.01);小鼠结肠和胃病理损伤明显改善(P<0.01)。脾阳虚泄泻模型小鼠出现明显的肠道菌群紊乱,灶心土给药后可显著降低模型小鼠肠道丹毒丝菌科和异杆菌属,提高毛螺旋菌科和Lachnospiraceae_K4A136_roup属的相对丰度。结论:灶心土对脾阳虚泄泻模型小鼠有止泻、减少胃肠病理损伤的作用,其温脾止泻机制与肠道菌群密切相关。

Objective:To study the therapeutic effect of Zaoxintu(灶心土)on diarrhea mouse model with Piyangxu(脾阳虚)syndrome.Methods:Sixty mice were randomly assigned into the control group,model group,montmorillonite suspension(1.5 g/kg)group,and Zaoxintu(灶心土)(ZXT 4,8,and 12 g/kg)groups.Except for the control group,all other mice were treated by"diet disorder+Kuhanxiexia(苦寒泻下)"method to establish the diarrhea model of Piyangxu(脾阳虚)syndrome.On day 11,the mice were administered by gavage with corresponding drugs or normal saline once daily for 7 days.The changes of mental state and body weight of mice before and after administration were observed,and the diarrhea rate and loose stool rate were calculated.The pathological changes of colon and stomach were observed via HE staining and scored according to relevant standards.The 16S rRNA gene sequencing was carried out to determine the diversity,abundance,and composition of gut microbiota and then the differential microbial groups were identified.Results:Compared with the model group,1.5 g/kg montmorillonite and 12 g/kg ZXT increased the body weight(P<0.01),decreased the diarrhea rate and loose stool rate(P<0.01),and alleviated the pathological damage of colon and stomach(P<0.01).The disorder of gut microbiota was observed in the diarrhea mouse model with Piyangxu(脾阳虚)syndrome.Compared with the model group,ZXT treatment reduced the relative abundance of Erysipelotrichaceae and Allobaculum and increased that of Lachnospiraceae and Lachnospiraceae_NK4A136_group.Conclusions:Zaoxintu(灶心土)has anti-diarrhea effect and can alleviate the gastrointestinal pathological damage in diarrhea mouse model with Piyangxu(脾阳虚)syndrome by regulating gut microbiota.