基于真实世界数据挖掘及实验探讨酒精性肝病肝郁脾虚证的用药规律及作用机制*

作者：王雅欣1，刘思彤2，吴 颖1，宋思琦1，高 翔1，宋 瑾1，3，田敬华1，刘 汶1，张会存1,3

单位：1.首都医科大学附属北京中医医院，北京 100010； 2.北京中医药大学，北京 100029；3.北京市中医药研究所，北京 100010

引用：引用：王雅欣，刘思彤，吴颖，宋思琦，高翔，宋瑾，田敬华，刘汶，张会存.基于真实世界数据挖掘及实验探讨酒精性肝病肝郁脾虚证的用药规律及作用机制[J].中医药导报,2025,31(5):56-62,70.

DOI：10.13862/j.cn43-1446/r.2025.05.010

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摘要：

目的：基于真实世界数据，探究中医药治疗酒精性肝病肝郁脾虚证的用药规律，分析核心药物可能的作用机制，并通过体外实验进行验证。方法：筛选酒精性肝病肝郁脾虚证患者处方信息，建立数据库，使用Microsoft Excel 2019、IBM SPSS Statistics 22.0和SPSS Molder 18.0软件进行频数统计、性味归经统计、关联规则和聚类分析，筛选出核心药物。应用中药系统药理学数据库与分析平台（TCMSP）获取核心药物的有效成分及靶点，GeneCards数据库获取酒精性肝病疾病靶点，应用Cytoscape 3.8.0软件构建“药物-成分-靶点”网络，应用String平台进行PPI网络分析，应用Metascape平台进行基因本体（GO）与京都基因与基因组百科全书（KEGG）通路富集分析。采用MTT法筛选核心药物最佳干预浓度，将大鼠肝星状细胞（HSC-T6）分为正常对照组、模型组、低剂量组、中剂量组、高剂量组，采用不同浓度含药培养基对酒精诱导活化后的HSC-T6细胞进行干预，MTT法检测细胞活力，RT-PCR法检测各组HSC-T6细胞中α-SMA mRNA、Col-1 mRNA及Col-3 mRNA的表达水平及PI3K/Akt信号通路中PI3K mRNA、Akt mRNA的表达水平。结果：治疗酒精性肝病肝郁脾虚证中药药性以寒、温为主，药味以甘、苦、辛味为主，归经以肝、胃经为主。共获得27组关联药物，药物聚类6组。根据药物应用频数筛选核心药物为白术、黄芪、泽兰、茯苓。有效成分共26种，药物与疾病共同靶点100个。GO分析获得生物过程239个，细胞成分33个，分子功能126个，KEGG通路富集分析主要与PI3K/Akt信号通路、JAK/STAT信号通路、NF-κB信号通路等有关。MTT结果显示，低剂量组、中剂量组、高剂量组均能抑制酒精诱导的HSC-T6细胞活化增殖（P<0.05）。RT-PCR结果显示，低、中、高剂量组能够显著下调酒精诱导活化的HSC-T6细胞中α-SMA mRNA、Col-1 mRNA、Col-3 mRNA、PI3K mRNA及Akt mRNA表达水平（P＜0.05）。结论：治疗酒精性肝病肝郁脾虚证的核心药物组合“白术-黄芪-泽兰-茯苓”能够通过调控PI3K/Akt信号通路抑制酒精性肝病进展。

关键词：酒精性肝病；肝郁脾虚证；用药规律；网络药理学；肝纤维化；大鼠肝星状细胞；PI3K/Akt信号通路

Abstract：

Objective: To explored the medication patterns of traditional Chinese medicine (TCM) in treating alcoholic liver disease (ALD) with liver depression and spleen deficiency based on real world data, analyze the possible mechanisms of core Chinese medicine, and verify the mechanisms through in vitro experiments. Methods: The prescription information for patients with ALD of liver depression and spleen deficiency was screened. A database was set up and Microsoft Excel 2019, IBM SPSS Statistics 22.0 and SPSS Molder 18.0 software were used for frequency statistics, property analysis, association rules analysis and cluster analysis. Then the core Chinese medicine was screened. Traditional Chinese medicine systems pharmacology database and analysis platform (TCMSP) was used to obtain the active components and targets of core drugs. GeneCards database was used to obtain the targets related to ALD. Cytoscape 3.8.0 software was used to construct a "drugs-components-targets" network. String platform was used for protein-protein interaction (PPI) network analysis, and Metascape platform was used for gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis. The MTT method was used to screen the optimal intervention concentration of drugs. HSC-T6 cells were divided into normal control group, model group, low dose group, medium dose group and high dose group. Different concentrations of drug containing culture medium were used to intervene in alcohol induced activation of HSC-T6 cells. MTT method was used to detect cell viability. RT-PCR method was used to detect the expression levels of α-SMA mRNA, Col-1 mRNA, and Col-3 mRNA in each group of HSC-T6 cells, as well as the expression levels of PI3K mRNA and Akt mRNA in PI3K/Akt signaling pathway. Results: The Chinese medicine used to treat ALD with liver depression and spleen deficiency was mainly cold and warm in nature, sweet, bitter, and spicy in taste, and mainly based on the liver and stomach meridians. A total of 27 groups of associated Chinese medicine were obtained, with 6 clusters. Based on the frequency of drug application, the core Chinese medicine selected were Baizhu (atractylodes macrocephala koidz), Huangqi (hedysarum multijugum maxim), Zelan (lycopi herba), and Fuling (poria cocos wolf), with a total of 26 active ingredients and 100 common targets both in drugs and disease. GO analysis obtained 239 biological processes, 33 cellular components, and 126 molecular functions. KEGG pathway enrichment analysis was mainly related to PI3K/Akt signaling pathway, JAK/STAT signaling pathway, and NF-κB signaling pathway. The MTT results showed that the low, medium, and high dose groups could inhibit alcohol induced activation and proliferation of HSC-T6 cells (P<0.05). The RT-PCR results showed that the low, medium, and high dose group could significantly down-regulate the expression levels of α-SMA mRNA, Col-1 mRNA, Col-3 mRNA, PI3K mRNA and Akt mRNA in alcohol induced activated HSC-T6 cells (P<0.05). Conclusion: The core Chinese medicine combination "Baizhu (atractylodes macrocephala koidz)-Huangqi (hedysarum multijugum maxim)-Zelan (lycopi herba)-Fuling (poria cocos wolf)" can inhibit the progression of ALD with liver depression and spleen deficiency by regulating the PI3K/Akt signaling pathway.

Key words：alcoholic liver disease; liver depression and spleen deficiency syndrome; medication patterns; network pharmaclolgy; liver fibrosis; rat hepatic stellate cells; PI3K/Akt signaling pathway

发布时间：2025-12-30

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