首页|补中益气汤经肠道菌群的调控改善脾虚证的作用机制研究

补中益气汤经肠道菌群的调控改善脾虚证的作用机制研究

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探讨补中益气汤通过调控肠道菌群改善脾虚证的作用机制。采用人源粪菌移植(脾虚证患者粪菌移植)和苦寒泻下(番泻叶10 g·kg-1)法建立脾虚证小鼠模型,给予补中益气汤(8。58 g·kg-1)治疗5 d,并以伪无菌(大剂量联合抗生素)和健康人粪菌移植小鼠为对照;通过酶联免疫法检测肠组织免疫球蛋白(IgA)、白细胞介素(IL)-2、IL-1β、干扰素(IFN)-γ、肝瘤坏死因子(TNF)-α、5-羟色胺(5-HT)变化,流式细胞仪测定肠组织CD8+/CD3+细胞比例,16S rRNA基因高通量测序和qPCR检测肠道菌群组成及含量变化,相关性分析探讨肠道菌群在改善脾虚证中介导作用。结果表明,补中益气汤显著升高粪菌移植和番泻叶2种脾虚证小鼠肠组织IgA,降低其IL-1β、TNF-α、5-HT和CD8+/CD3+细胞比例,对IL-2和IFN-y在2种脾虚证小鼠肠组织的变化具有双向调节作用。补中益气汤可抑制番泻叶脾虚证小鼠肠道菌群的过度生长,降低群落丰富度;对2种脾虚证小鼠紊乱的肠道菌群结构均有改善作用。与正常小鼠菌群的比较分析表明,Algoriphagus、Mycobacterium、CL500_29_marine_group为2种脾虚证模型的共有差异菌属;Acinetobacter、Parabacteroides、Ruminococcus为粪菌移植脾虚证模型特有差异菌属;Sphingorhabdiis、Lactobacillus、Anaeroplasma为番泻叶脾虚证模型特有差异菌属,补中益气汤对上述菌属均有明显调节作用。qPCR分析表明,补中益气汤可明显升高2种脾虚证模型小鼠粪便中Akkermansia muciniphila含量,降低Bacteroides uniformis含量。相关性分析发现,在不同方式诱导的脾虚证动物模型中,上述肠道免疫相关细胞因子与特征性肠道微生物具有明显相关性,其中IL-1β在2个脾虚证模型中与Acinetobacter和CL500_29_marine_group均显著正相关,而2种脾虚证模型中肠组织IL-2和IFN-γ水平不同可能与其不同菌群结构有关。综上,补中益气汤可调节不同脾虚证动物模型中紊乱的肠道菌群结构,改善其肠道免疫状态,这可能是其治疗脾虚证的作用机制之一。
Buzhong Yiqi Decoction ameliorates spleen deficiency syndrome by regulating gut microbiota
This study aims to decipher the mechanism of Buzhong Yiqi Decoction(BZYQD)in the treatment of spleen deficiency syndrome via gut microbiota.The mouse models of spleen deficiency syndrome were established by fecal microbiota transplantation(FMT,from patients with spleen deficiency syndrome)and administration of Sennae Folium(SF,10 g·kg-1),respectively,and trea-ted with BZYQD for 5 d.The pseudosterile mice(administrated with large doses of antibiotics)and the mice transplanted with fecal bacteria from healthy human were taken as the controls.The levels of IgA,interleukin(IL)-2,IL-1β,interferon(IFN)-γ,tumor nec-rosis factor-alpha(TNF-α),and 5-hydroxytryptamine(5-HT)in the intestinal tissue of two models were measured by enzyme-linked immunosorbent assay,and the CD8+/CD3+ratio was determined by flow cytometry.The composition and changes of the gut microbiota were determined by 16S rRNA high-throughput sequencing and qPCR.Furthermore,the correlation analysis was performed to study the mediating role of gut microbiota in the treatment.The results showed that BZYQD elevated the IgA level,lowered the IL-1β,TNF-α,and 5-HT levels,and decreased the CD8+/CD3+ratio in the intestinal tissue of the two models.Moreover,BZYQD had two-way regu-latory effects on the levels of IL-2 and IFN-y.BZYQD inhibited the overgrowth and reduced the richness of gut microbiota in the SF model,and improved the gut microbiota structure in the two models.Algoriphagus,Mycobacterium,and CL500_29_marine_group were the common differential genera in the two models compared with the control.Acinetobacter,Parabacteroides,and Ruminococcus were the differential genera unique to the FMT model,and Sphingorhabdus,Lactobacillus,and Anaeroplasma were the unique differential genera in the SF model.BZYQD was capable of regulating all these genera.The qPCR results showed that BZYQD increased the rela-tive abundance of Akkermansia muciniphila and decreased that of Bacteroides uniformis in the two models.The correlation analysis re-vealed that the levels of above intestinal cytokines were significantly correlated with characteristic gut microorganisms in different mo-dels.The IL-1β level had a significantly positive correlation with Acinetobacter and CL500_29_marine_group in the two models,while the different levels of IL-2 and IFN-γ in the two models may be related to its different gut microbiota structures.In conclusion,BZYQD could regulate the disordered gut microbiota structure in different animal models of spleen deficiency syndrome to improve the intestinal immune status,which might be one of the mechanisms of BZYQD in treating spleen deficiency syndrome.

Buzhong Yiqi Decoctionspleen deficiency syndromefecal microbiota transplantationSennae Foliumanimal model

于涵川、孟杨杨、王恩康、袁建业、彭颖、李晓波

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上海交通大学药学院,上海 200240

上海中医药大学 附属龙华医院脾胃病研究所,上海 200032

补中益气汤 脾虚证 粪菌移植 番泻叶 动物模型

国家自然科学基金国家科技支撑计划重点项目(十一五)

818039902006BAI08B05-02

2024

中国中药杂志
中国药学会

中国中药杂志

CSTPCD北大核心
影响因子:1.718
ISSN:1001-5302
年,卷(期):2024.49(4)
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