探讨中药致胆汁淤积型肝损伤的特点及调控机制

Characteristics and Regulatory Mechanisms of Cholestatic Liver Injury Caused by Chinese Herbal Medicines

摘要:

目的 基于数据挖掘、网络药理学及分子对接技术探讨中药导致胆汁淤积型肝损伤（CLI）的特点及其相关的调控路径。方法 利用中国知网、PubMed等数据库检索建库至2024年关于中药导致的相关文献，规范信息后汇总并进行聚类分析；运用中药系统药理数据分析平台（TCMSP）、中医药整合药理学研究平台（TCMIP），GeneCards、在线人类孟德尔遗传数据库（OMIM）、DisGeNET数据库检索核心药物靶点及相关靶点；利用韦恩图绘制核心药物与CLI的交集靶点；利用Cytoscape 3.10.2构建交集靶点的蛋白相互作用（PPI）网络图；通过DAVID数据库对核心靶点进行基因本体（GO）和京都基因与基因组百科全书（KEGG）富集分析；最后用AutoDock Vina进行分子对接。结果 该研究共筛选出符合条件的文献849篇，统计出39味可导致胆汁淤积的中药及其单体成分64个。按2020年版《中华人民共和国药典》进行归类，其中报道频次由高到低依次为清热药、解表药、活血化瘀药、泻下药、化痰止咳平喘药、补益药、祛风湿药、温里药、利水渗湿药、理气药、止血药、攻毒杀虫止痒药、收涩药、化湿药、安神药。聚类分析发现清热药致CLI的研究报道数量最多，占比39.69%。其中，清热药中报道较多的是栀子（92篇，占比10.84%）、黄芩（76篇，占比8.95%）、苦参（69篇，占比6.95%）。网络毒理学研究可知被报道导致CLI的中药引起胆汁淤积的核心靶点主要包括肿瘤坏死因子（TNF）、过氧化物酶体增殖物激活受体α（PPARA）、法尼醇X受体（FXR）、谷丙转氨酶2抗体（GPT2）、超氧化歧化酶1（SOD1）、干扰素γ（IFN-γ）、白细胞介素-6（IL-6）、CD36、载脂蛋白A1（APOA1）、血管紧张素转化酶（ACE）、细胞色素P450 3A4酶（CYP3A4）、蛋白激酶B1（Akt1）、APOB、白蛋白（ALB）、ATP结合盒转运蛋白B4（ABCB4）、SLC10A1、雌激素受体α（ESR1）、信号转导与转录激活因子1（STAT1）、肌动蛋白（ACTB）、内皮素1（EDN1）、ABCG2、过氧化物酶体增殖物激活受体γ（PPARG）等，涉及的信号通路有胆汁分泌、ABC转运体、类固醇激素的生物合成、DNA加合物、药物代谢、细胞色素P450、过氧化物酶体、初级胆汁酸的生物合成、视黄醇代谢、Toll样受体等。分子对接结果表明，被高频报道可导致胆汁淤积肝损伤的清热药的有效成分（如黄芩苷和黄连素等）与ABCG2、IFN-γ、EDN1、IL-6、SOD1等靶点结合良好，结合能在-13~-9 kcal·mol^-1，调控途径与转运体、微血管功能调节、炎症和氧化应激等高度相关，这与聚类分析的结果一致。结论 根据文献报道及数据库分析，致CLI的中药主要集中在清热药，且核心作用靶点为ABCG2、IFN-γ、EDN1、IL-6、SOD1等，调控途径主要与转运体、微血管功能调节、炎症、氧化应激等有关。

Abstract:

Objective To explore the characteristics and regulatory mechanisms of cholestatic liver injury （CLI） caused by traditional Chinese medicine based on data mining， network pharmacology， and molecular docking.Method China National Knowledge Infrastructure and PubMed were searched for the relevant literature on CLI caused by traditional Chinese medicine from inception to 2024， and the information was standardized， summarized， and analyzed by cluster analysis. The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine （TCMIP）， GeneCards， Online Mendelian Inheritance in Man （OMIM）， and DisGeNET were used to retrieve the active ingredients and targets of core medicines. The Venn diagram was established to map the common targets shared by the core medicines and CLI. Cytoscape 3.10.2 was used to construct the protein-protein interaction （PPI） network of the common targets. DAVID was used for gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment of the core targets. Finally， molecular docking was performed by AutoDock Vina.Result A total of 849 eligible articles were included in this study， from which 64 active ingredients of 39 herbal medicines that can cause cholestasis were counted and categorized according to the 2020 edition of the Pharmacopoeia of the People's Republic of China. The frequency of the medidicnes followed a descending order of heat-clearing medicines， exterior-releasing medicines， blood-activating and stasis-resolving medicines， purgative medicines， phlegm-resolving and cough- and dyspnea-relieving medicines， tonics， wind- and dampness-expelling medicines， interior-warming medicines， urination-promoting and dampness-draining medicines， Qi movement-regulating medicines， hemostatics， toxin-removing， worm-killing， and itch-relieving medicines， astringent medicines， dampness-eliminating medicines， and tranquiling medicines. The cluster analysis revealed that the reports of CLI caused by heat-clearing medicines accounted for the highest proportion of 39.69%. Among the heat-clearing medicines， Gardeniae Fructus （92 articles， accounting for 10.84%）， Scutellariae Radix （76 articles， 8.95%）， and Sophorae Flavescentis Radix （69 articles， 6.95%） were frequently reported. The core targets of cholestasis induced by Chinese herbal medicines reported to cause CLI mainly included tumor necrosis factor （TNF）， peroxisome proliferator activated receptor alpha （PPARA）， farnesoid X receptor （FXR）， glutamic-pyruvic transaminase 2 （GPT2）， superoxide dismutase 1 （SOD1）， interferon gamma （IFN-γ）， interleukin-6（IL-6）， CD36， Apolipoprotein A1（APOA1）， Angiotensin converting enzyme（ACE）， Cytochrome P450 3A4 enzyme（CYP3A4）， Protein kinase B1（Akt1）， APOB， albumin（ALB）， ATP binding cassette transporter A4（ABCB4）， SLC10A1， Estrogen Receptor alpha （ESR1）， signal transducer and activator of transcription1（STAT1）， β-actin（ACTB）， Endothelin 1（EDN1）， ABCG2， and peroxisome proliferator activated receptor gamma（PPARG）. The signaling pathways involved included bile secretion， ABC transporter， steroid biosynthesis， DNA adducts， drug metabolism， cytochrome P450， peroxisomes， primary bile acid biosynthesis， retinol metabolism， and Toll-like receptor. The molecular docking results showed that the active ingredients （e.g.， baicalin and berberine） of the heat-clearing medicines reported by high frequency to cause CLI had high binding affinity to the targets including ABCG2， IFN-γ， EDN1， IL-6 and SOD1， with the binding energy in the range of -13 kcal·mol^-1 to -9 kcal·mol^-1， and the regulatory pathways were highly correlated with transporters， microvascular function regulation， inflammation， and oxidative stress， which was consistent with the cluster analysis.Conclusion The available reports about the Chinese herbal medicines causing CLI mainly focused on heat-clearing medicines， and the core targets included ABCG2， IFN-γ， EDN1， IL-6， and SOD1. The regulatory pathways were mainly related to transporters， microvascular function regulation， inflammation， and oxidative stress.