<i>In silico</i> and <i>in vivo</i> demonstration of the regulatory mechanism of Qi-Ge decoction in treating NAFLD

Nonalcoholic fatty liver disease (NAFLD), a chronic and progressive liver disease, often causes steatosis and steatohepatitis. Qi-Ge decoction (QGD) shows a good effect against NAFLD in the clinic. But the molecular mechanism for QGD in improving NAFLD is unknown. This study explored the molecular mechanism of QGD in NAFLD model rats using comprehensive network pharmacology, molecular docking and <i>in vivo</i> verification strategies. Active components and targets of QGD were obtained from public database. The overlapped genes between QGD and NAFLD targets were analyzed by enrichment analysis. Active components and targets were used to predict molecular docking analysis. Finally, seven key targets were screened out and the gene expression were verified in the NAFLD rat’s liver tissues after QGD treatment. Fifty-eight common QGD therapeutic targets were associated with NAFLD. Molecular docking demonstrated that seven targets had strong binding ability for the corresponding active ingredients. GO analysis identified 18 biological process entries, which were mainly related to regulation of lipid storage, lipid localization and peptide transport. KEGG analysis identified multiple signaling pathways, which were mainly associated with tumor necrosis factor signaling and NAFLD. <i>In vivo</i> data confirmed that the effect of QGD in the treatment of NAFLD was mainly exerted through improving liver steatosis and inflammatory cell infiltration. Additionally, QGD upregulated the expression of MAPK8 and ESR1 and downregulated the transcriptional expression of IL6, VEGFA, CASP3, EGFR and MYC. These targets may affect lipid metabolism by regulating lipid storage and inflammation. The integration of results obtained <i>in silico</i> and <i>in vivo</i> indicated that QGD regulates multiple targets, biological processes and signaling pathways in NAFLD, which may represent a complex molecular mechanism by which QGD improves NAFLD.Key messagesQGD intervention is related to multiple biological processes such as inflammation, oxidation and cell apoptosis in NAFLD.Lipid and atherosclerosis, TNF signaling pathway, IL-17 signaling pathway, non-alcoholic fatty liver disease and AGE-RAGE signaling pathway in diabetic complications are the main pathways for QGD intervention NAFLD.The active components of QGD can form good binding with relevant target proteins through intermolecular forces, exhibiting excellent docking activity. QGD intervention is related to multiple biological processes such as inflammation, oxidation and cell apoptosis in NAFLD. Lipid and atherosclerosis, TNF signaling pathway, IL-17 signaling pathway, non-alcoholic fatty liver disease and AGE-RAGE signaling pathway in diabetic complications are the main pathways for QGD intervention NAFLD. The active components of QGD can form good binding with relevant target proteins through intermolecular forces, exhibiting excellent docking activity.非酒精性脂肪性肝病（Nonalcoholic fatty liver disease, NAFLD）是一种慢性进展性肝脏疾病，常引发脂肪变性及脂肪性肝炎。芪葛汤（Qi-Ge decoction, QGD）在临床中展现出良好的抗NAFLD疗效，但其改善NAFLD的分子机制尚不明确。本研究采用整合网络药理学、分子对接及体内（in vivo）验证策略，探究了QGD治疗NAFLD模型大鼠的分子机制。首先从公共数据库中获取QGD的活性成分与作用靶点；随后对QGD与NAFLD的交集基因进行富集分析；并利用活性成分与靶点开展分子对接预测。最终筛选出7个关键靶点，并在QGD干预后的NAFLD大鼠肝组织中验证了这些靶点的基因表达水平。本研究共获得58个QGD治疗NAFLD的共同靶点。分子对接结果显示，7个核心靶点与对应活性成分具有较强的结合能力。GO分析共鉴定得到18条生物学过程条目，主要涉及脂质储存调控、脂质定位及肽转运过程。KEGG分析筛选得到多条信号通路，主要与肿瘤坏死因子信号通路及非酒精性脂肪性肝病相关。体内实验数据证实，QGD治疗NAFLD的作用主要通过改善肝脏脂肪变性及炎性细胞浸润实现。此外，QGD可上调MAPK8与ESR1的表达，同时下调IL6、VEGFA、CASP3、EGFR及MYC的转录水平。上述靶点可能通过调控脂质储存与炎症反应影响脂质代谢。对计算机预测（in silico）与体内实验结果的整合分析表明，QGD可通过调控NAFLD进程中的多个靶点、生物学过程及信号通路发挥治疗作用，这可能代表了QGD改善NAFLD的复杂分子机制。 关键信息 QGD干预NAFLD的作用与炎症、氧化应激及细胞凋亡等多种生物学过程密切相关。脂质与动脉粥样硬化、TNF信号通路、IL-17信号通路、非酒精性脂肪性肝病及糖尿病并发症中的AGE-RAGE信号通路是QGD干预NAFLD的主要通路。QGD的活性成分可通过分子间作用力与相关靶蛋白形成良好结合，展现出优异的对接活性。 QGD干预NAFLD的作用与炎症、氧化应激及细胞凋亡等多种生物学过程密切相关。脂质与动脉粥样硬化、TNF信号通路、IL-17信号通路、非酒精性脂肪性肝病及糖尿病并发症中的AGE-RAGE信号通路是QGD干预NAFLD的主要通路。QGD的活性成分可通过分子间作用力与相关靶蛋白形成良好结合，展现出优异的对接活性。