铁皮石斛对脾虚便秘小鼠肠道菌群多样性的影响

Effect of Dendrobium officinale on the diversity of intestinal microbiota in mice with spleen deficiency constipation

目的探讨铁皮石斛对脾虚便秘小鼠肠道菌群的影响,为铁皮石斛对脾虚便秘的疗效机制及临床组方提供依据。方法选取12只SPF级昆明小鼠随机分为正常组、模型组、铁皮石斛组和枳术汤组,每组3只。造模阶段正常组灌胃无菌水,模型组和治疗组采用番泻叶水煎液灌胃7 d后,限制饮食饮水8 d,建立脾虚便秘模型。造模成功后,正常组和模型组灌胃无菌水,治疗组分别以铁皮石斛和枳术汤水煎液干预5 d。治疗结束后,收集各组小鼠盲肠内容物样本进行16S rDNA高通量测序。结果脾虚便秘小鼠排便量减少,粪便细小,干燥,质硬,呈黄色。铁皮石斛和枳术汤分别治疗后,小鼠排便量增加,粪便软硬适中,呈黑褐色。测序分析发现,正常组与铁皮石斛组的共有OTU数最多,与枳术汤组的共有OTU数最少。α-多样性指数与PCA结果表明,铁皮石斛组菌属结构较其余两组更接近正常组小鼠。物种相对丰度及组成结果表明,铁皮石斛增加了脾虚便秘小鼠双歧杆菌属相对丰度,抑制了乳杆菌属、拟杆菌属相对丰度,但对菌群丰度的调节作用不及枳术汤;LEfSe分析与随机森林分析鉴定出布劳特菌属和活泼瘤胃球菌是铁皮石斛组与其他组别的差异菌群。结论铁皮石斛能恢复脾虚便秘小鼠肠道菌群的多样性,可调节肠道菌群相对丰度,布劳特菌属和活泼瘤胃球菌可能是铁皮石斛干预脾虚便秘小鼠的肠道关键指示菌。

Objective To observe the effects of Dendrobium officinale(DO)on the intestinal microbiota of mice with spleen-deficiency constipation(SDC)and provide a basis for its therapeutic mechanism and clinical prescription formulation.Methods Twelve Kunming mice were divided into four groups:normal controls,models,DO,and Zhizhu decoction(n=3 each).The normal control group received sterile water;the other groups received senna leaf decoction for 7 days,followed by 8 days of diet and water restriction to induce SDC.After modeling,the treatment groups received DO or Zhizhu decoction respectively for 5 days.Cecal samples were then collected for 16S rDNA sequencing.Results In mice with SDC,fecal output was diminished,resulting in small,hard and dry feces.Treatment with DO and Zhizhu decoction improved fecal output,yielding softer,dark brown feces.Operational taxonomic unit(OTU)analysis revealed the normal control group had the most OTUs in common with the DO group,and the fewest with the Zhizhu group.In terms of α-diversity and principal component analysis(PCA),the DO group closely resembled the normal control group compared to the other treatments.Further analysis showed that DO increased the relative abundance of Bifidobacteria while inhibiting Lactobacillus and Pseudomonas,albeit less effectively than Zhizhu decoction.LEfSe and random forest analyses identified Blautia and Ruminococcus gnavus as a distinctive bacterium in the DO group.Conclusion DO was more effective than Zhizhu decoction for restoring gut microbial diversity and modulating key bacterial genera in mice with SDC.However,it was not as potent as Zhizhu in regulating overall microbiota abundance.Blautia and R.gnavus may serve as a potential biomarker in DO intervention for SDC.