目的 基于网络药理学和实验验证探讨甘草Glycyrrhizae Radix et Rhizoma防治非酒精性脂肪肝病（nonalcoholic fatty liver disease，NAFLD）及肥胖的活性成分、作用靶点及潜在机制。方法 采用TCMSP数据库结合已发表文献筛选获得甘草主要活性化学成分及其作用靶点，GeneCards数据库获取NAFLD及肥胖靶点，分别用String平台、Cytoscape 3.9.1软件分析并构建蛋白质-蛋白质相互作用（protein-protein interaction，PPI）网络。借助Metascape平台分析交集靶点并进行基因本体（gene ontology，GO）功能及京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）通路富集分析，采用Cytoscape 3.9.1软件构建“甘草成分-疾病靶点-通路”网络，进一步采用分子对接验证预测的关键成分与靶点之间的结合能力与亲和力，通过动物实验证实关键成分对靶点通路的调控作用。结果 甘草主要通过核心活性成分槲皮素、甘草酸、山柰酚、柚皮素和芒柄花素等作用于前列腺素G/H合酶2（prostaglandin G/H synthase 2，PTGS2）、雌激素受体1（estrogen receptor 1，ESR1）、过氧化物酶体增殖激活受体γ（peroxisome proliferator-activated receptor gamma，PPARG）、一氧化氮合酶2（nitric oxide synthase 2，NOS2）及糖原合成酶激酶3β（glycogen synthase kinase-3 beta，GSK3B）等关键靶点蛋白，经脂质与动脉粥样硬化、胰岛素抵抗、腺苷酸活化蛋白激酶（adenosine monophosphate-activated protein kinase，AMPK）信号通路、PPAR信号通路、T细胞受体信号等通路发挥防治NAFLD及肥胖的功效。活性成分甘草酸可以缓解高脂饮食小鼠肝脏脂肪变性，显著减轻小鼠体质量增加（P<0.05），明显改善血脂及肝功能水平（P<0.05、0.01），同时显著增加肝脏视黄酸受体α（retinoic acid receptor RXR-alpha，RXRA）、PPARG及胆固醇7α-羟化酶（cholesterol 7α-hydroxylase，CYP7A1）的mRNA及蛋白表达（P<0.05、0.01）。结论 甘草通过多成分、多靶点、多途径干预NAFLD及肥胖，其主要活性成分甘草酸能通过激活高脂饮食小鼠肝脏RXRA/PPARG/CYP7A1信号通路发挥抗NAFLD及肥胖药效作用。

Objective To explore the active ingredients, targets and potential mechanism of Gancao (Glycyrrhizae Radix et Rhizoma) in the prevention and treatment of non-alcoholic fatty liver disease (NAFLD) and obesity based on network pharmacology and experimental verification. Methods The main chemical active constituents and their action targets of Glycyrrhizae Radix et Rhizoma were screened by TCMSP database combined with published literature, and the targets of NAFLD and obesity were obtained from GeneCards database. The protein-protein interaction (PPI) network was following analyzed and constructed using String platform and Cytoscape 3.9.1 software, respectively. Then, the intersection targets were analyzed by Metascape platform, and gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were performed. In addition, Cytoscape 3.9.1 software was used again to construct the "component of Glycyrrhizae Radix et Rhizoma-disease target-pathway" network. Molecular docking was further used to verify the binding ability and affinity between the predicted key components and targets. And finally, the regulatory effect of key components on the target pathway was confirmed by animal experiments. Results Glycyrrhizae Radix et Rhizoma mainly acted on key targets such as prostaglandin G/H synthase 2 (PTGS2), estrogen receptor 1 (ESR1), peroxisome proliferator-activated receptor gamma (PPARG), nitric oxide synthase 2 (NOS2) and glycogen synthase kinase-3 beta (GSK3B) through core active components like quercetin, glycyrrhizic acid, kaempferol, naringin and formononetin. And it might prevent and treat NAFLD and obesity via lipid and atherosclerosis, insulin resistance, adenosine monophosphate-activated protein kinase (AMPK) signaling pathway, PPAR signaling pathway, T cell receptor signaling and other pathways. The active ingredient glycyrrhizic acid could alleviate liver steatosis, dramatically reduce weight gain (P < 0.05), observably improve blood lipid level as well as liver function (P < 0.05, 0.01), and significantly increase both hepatic mRNA and protein expressions of retinoic acid receptor RXR-alpha (RXRA), PPARG and cholesterol 7α-hydroxylase (CYP7A1) in high fat diet mice (P < 0.05, 0.01). Conclusion Glycyrrhizae Radix et Rhizoma could effectively intervene in NAFLD and obesity with the characteristics of multi-component, multi-target and multi-pathway. And its main active ingredient glycyrrhizic acid could play a role in anti-NAFLD and obesity by activating hepatic RXRA/PPARG/CYP7A1 signaling pathway in high fat diet mice.

国家自然科学基金资助项目（82100574）；天津市卫生健康科技项目（TJWJ2022QN010）