基于代谢组学探讨益气养阴解毒化瘀方对肝硬化轻微型肝性脑病肠道代谢物的影响

姜婷婷; 张群; 王宪波; 江宇泳; 刘晓利; 于浩; 杨志云

doi:10.12449/JCH250312

基于代谢组学探讨益气养阴解毒化瘀方对肝硬化轻微型肝性脑病肠道代谢物的影响

DOI: 10.12449/JCH250312

首都医科大学附属北京地坛医院中西医结合中心，首都医科大学基础-临床联合实验室，北京 100015

基金项目:

国家自然科学基金 (82104781);

北京市属医院科研培育计划 (PZ20241802);

北京市医院管理中心“登峰”人才培养计划 (DFL20191803);

高层次公共卫生技术人才建设项目培养计划 (Discipline Leader-02-16)

伦理学声明：本研究方案于2024年3月7日经首都医科大学附属北京地坛医院伦理委员会审批，批号：京地伦科字（2024）第（015）-01号，所有入组患者及家属均签署知情同意书。

利益冲突声明：本文不存在任何利益冲突。

作者贡献声明：姜婷婷、张群、江宇泳、王宪波负责课题设计，资料分析，撰写论文；于浩、刘晓利参与收集数据，修改论文；杨志云负责拟定写作思路，指导撰写文章并最后定稿。

详细信息

通信作者:
杨志云， yangzhiyun2016@163.com （ORCID： 0000-0002-1522-0749）

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出版历程
- 收稿日期: 2024-07-08
- 录用日期: 2024-08-12
- 出版日期: 2025-03-25

Effect of Yiqi Yangyin Jiedu Huayu prescription on intestinal metabolites in liver cirrhosis with minimal hepatic encephalopathy： A study based on metabolomics

Department of Integrated Traditional Chinese and Western Medicine，Beijing Ditan Hospital，Capital Medical University，Laboratory for Clinical Medicine，Capital Medical University，Beijing 100015，China

Research funding:

National Natural Science Foundation of China (82104781);

Research and Cultivation Program of Beijing Municipal Hospital (PZ20241802);

Beijing Hospital Management Center “Peak” talent training program (DFL20191803);

High-level Public Health Technical Personnel Construction Project Training Program (Discipline Leader-02-16)

More Information

Corresponding author: YANG Zhiyun， yangzhiyun2016@163.com （ORCID： 0000-0002-1522-0749）

摘要

摘要: 目的通过肠道代谢组学探讨益气养阴解毒化瘀方对肝硬化轻微型肝性脑病（MHE）的作用。方法选取2024年3月—2024年5月于首都医科大学附属北京地坛医院住院且经肝性脑病心理测量评分诊断为MHE的肝硬化患者（n=11）作为研究对象，同时招募健康患者家属作为对照组（n=11）。通过采集对照组及益气养阴解毒化瘀方治疗前后MHE患者的粪便进行代谢组学分析，探索MHE患者的肠道代谢物特征，并通过人群队列探讨益气养阴解毒化瘀方对MHE患者肠道代谢的影响。计数资料两组间比较采用Fisher精确检验；符合正态分布的计量资料两组间比较采用成组t检验，同一组内治疗前后的比较采用配对t检验，非正态分布的计量资料两组间及同组内治疗前后的比较采用Wilcoxon秩和检验。结果 MHE组与对照组共检测出29种差异代谢物，差异代谢物主要为氨基酸类、有机酸类、有机胺类、碳水化合物类、脂肪酸类和维生素类，MHE组明显上调12种，下调17种，主要富集在鸟氨酸代谢通路、支链氨基酸代谢通路和芳香氨基酸代谢通路。MHE患者经益气养阴解毒化瘀方治疗前后共检测出80种差异代谢物，差异代谢物主要为碳水化合物类、有机酸类和氨基酸类，中药治疗后上调56种，下调24种，差异通路也主要富集在鸟氨酸代谢通路、支链氨基酸代谢通路和芳香氨基酸代谢通路。结论益气养阴解毒化瘀方可能通过调节MHE患者的肠道代谢发挥治疗作用。
- 肝硬化 /
- 肝性脑病 /
- 益气养阴解毒化瘀方 /
- 中医药疗法 /
- 代谢组学
Abstract: Objective To investigate the effect of Yiqi Yangyin Jiedu Huayu prescription on minimal hepatic encephalopathy in liver cirrhosis based on intestinal metabolomics. Methods A total of 11 patients with liver cirrhosis who were hospitalized in Beijing Ditan Hospital， Capital Medical University， from March to May 2024， and were diagnosed with MHE based on psychometric hepatic encephalopathy score were enrolled as subjects， and 11 healthy family members of the patients were enrolled as control group. Fecal samples were collected for metabolomics analysis from the control group and the patients with MHE before and after treatment with Yiqi Yangyin Jiedu Huayu prescription， and a population cohort study was conducted to investigate the effect of Yiqi Yangyin Jiedu Huayu prescription on intestinal metabolism of patients with MHE. The Fisher’s exact test was used for categorical data between two groups； the independent samples t-test was used for comparison of normally distributed continuous data between two groups， the paired t-test was used for comparision before and after treatment within the same group， and the Wilcoxon rank-sum test was used for comparison of non-normally distributed continuous data between two groups. Results A total of 29 differentially expressed metabolites were detected between the MHE group and the control group， mainly amino acids， organic acids， organic amines， carbohydrates， fatty acids， and vitamins， and there were 12 upregulated metabolites and 17 downregulated metabolites in the MHE group， which were mainly enriched in the metabolic pathways of ornithine， branched-chain amino acid， and aromatic amino acid. After the treatment with Yiqi Yangyin Jiedu Huayu prescription， 80 differentially expressed metabolites were detected in the patients with MHE， mainly carbohydrates， organic acids， and amino acids， and there were 56 upregulated metabolites and 24 downregulated metabolites， which were mainly enriched in the metabolic pathways of ornithine， branched-chain amino acid， and aromatic amino acid. Conclusion Yiqi Yangyin Jiedu Huayu prescription can exert a therapeutic effect on patients with MHE by regulating intestinal metabolism.
- Liver Cirrhosis /
- Hepatic Encephalopathy /
- Yiqi Yangyin Jiedu Huayu Prescription /
- Traditional Chinese Medicine Therapy /
- Metabolomics

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注：*P<0.05，**P<0.01，***P<0.001。

图 1 MHE组与对照组的主要代谢物分类组成

Figure 1. Classification and composition of major metabolites in MHE and control groups

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图 2 肠道差异性代谢物的单维统计分析

Figure 2. Unidimensional statistical analysis of intestinal differential metabolites

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注： Corr.Coeffs，代谢物与第一主成分相关性的相关系数。

图 3 肠道差异性代谢物的多维统计分析

Figure 3. Multidimensional statistical analysis of intestinal differential metabolites

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图 4 MHE患者的差异代谢通路

Figure 4. Differential metabolic pathways in MHE patients

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图 5 益气养阴解毒化瘀方治疗前后代谢物的PCA分析

Figure 5. PCA analysis of metabolites before and after Yiqi Yangyin Jidu Huayu prescription

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图 6 益气养阴解毒化瘀方治疗前后代谢物的PLS-DA分析

Figure 6. PLS-DA analysis of metabolites before and after Yiqi Yangyin Jidu Huayu prescription

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图 7 益气养阴解毒化瘀方治疗前后的主要代谢物分类组成

Figure 7. Classification and composition of main metabolites before and after treatment with Yiqi Yangyin Jiedu Huayu prescription

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图 8 益气养阴解毒化瘀方治疗前后肠道差异性代谢物的单维统计分析

Figure 8. Unidimensional statistical analysis of intestinal differential metabolices before and after treatment with Yiqi Yangyin Jiedu Huayu prescription

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图 9 益气养阴解毒化瘀方治疗前后肠道差异性代谢物的多维统计分析

Figure 9. Multidimensional statistical analysis of intestinal differential metabolites before and after treatment with Yiqi Yangyin Jiedu Huayu prescription

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图 10 益气养阴解毒化瘀方治疗前后的差异性代谢物种类热图

Figure 10. Heat map of different metabolites before and after treatment of Yiqi Yangyin Jiedu Huayu prescription

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图 11 益气养阴解毒化瘀方治疗前后的差异性代谢物散点图

Figure 11. Scatter diagram of differential metabolites before and after treatment of Yiqi Yangyin Jiedu Huayu prescription

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图 12 益气养阴解毒化瘀方治疗前后的差异性代谢通路分析

Figure 12. Analysis of different metabolic pathways before and after treatment with Yiqi Yangyin Jiedu Huayu prescription

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表 1 MHE组和对照组的基线资料

Table 1. Baseline characteristics of patients with MHE and control groups

项目	MHE组（n=11）	对照组（n=11）	统计值	P值
男/女（例）	8/3	7/4		0.833
年龄（岁）	61.00±11.17	60.00±8.44	t=0.237	0.815
BMI（kg/m²）	20.73±5.26	21.90±5.53	t=-0.513	0.613
NCT-A（s）	68.90±23.00	36.82±15.80	t=3.506	0.002
DST（分）	17.80±4.50	49.18±16.35	t=-6.101	<0.05

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表 2 12种上调的代谢物

Table 2. 12 up-regulated metabolites

代谢物分类	代谢物
脂肪酸类	柠苹酸
有机胺类	尿素、酪胺
有机酸类	乳酸、甲基丙二酸
氨基酸类	鸟氨酸
碳水化合物类	乳果糖、核糖核酸、木糖内酯、异麦芽糖
苯类	苯基乙胺
醇类	2，3-丁二醇

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表 3 17种下调的代谢物

Table 3. 17 down-regulated metabolites

代谢物分类	代谢物
脂肪酸类	油酸、亚油酸、己二酸、神经酸
有机酸类	酮亮氨酸
有机胺类	亚精胺
氨基酸类	肌氨酸
碳水化合物类	甘油酸、核糖
苯丙酸类	3-（3-羟基苯基）-3-羟基丙酸
维生素类	δ-生育醇、α-生育酚、γ-生育酚
胆汁酸类	二氯乙酸
内脂类	2-油酸单甘油酯
吡啶类	烟酸
咪唑类	咪唑丙烯酸

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表 4 益气养阴解毒化瘀方治疗前后肝功能的变化

Table 4. Changes in liver function before and after treatment with Yiqi Yangyin Jiedu Huayu prescription

指标	治疗前（n=11）	治疗后（n=11）	统计值	P值
ALT（U/L）	69.65±56.66	17.50±10.87	t=-3.532	0.005
AST（U/L）	53.80（31.90～87.50）	15.70（9.70～21.50）	t=-3.513	<0.001
TBil（μmol/L）	72.70（39.90～78.95）	25.50（19.28～30.43）	Z=-2.856	0.004
NH₃（μmol/L）	43.70±10.96	21.74±6.97	t=5.608	<0.001
Child-Pugh评分（分）	11.36±2.25	6.73±1.62	t=5.551	<0.001
MELD评分（分）	12.50（10.63～17.38）	5.00（2.83～9.67）	Z=-3.034	0.002
NCT-A（s）	68.90±23.00	46.18±16.48	t=2.093	0.049
DST（分）	17.80±4.50	23.45±5.43	t=-2.572	0.018

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