Introduction: The potential targets and action mechanism of Yiguanjian (YGJ) decoction in treating acute liver failure (ALF) were determined using network pharmacology, molecular docking and cell experiments.

Methods: The Pharmacology of Traditional Chinese Medicine Systems (TCMSP) and BATMAN-TCM databases were used to find YGJ decoction targets. The Genecard database was employed to predict targets associated with ALF. Cytascape software was utilised to visualise and select common targets along with establishing a protein–protein interaction (PPI) network. Core targets were screened and putative function was assessed via gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyzes. Molecular docking was performed using AutodockTools, PyMoL and Discovery Studio software to verify the correlation between the YGJ decoction and selected targets. MTT assay was conducted to determine the effect of the YGJ decoction on the viability of Huh-7 human hepatoma cells. The effects of the YGJ decoction on the expression of putative targets in Huh-7 cells were determined via western blot and quantitative real-time polymerase chain reaction analyzes.

Results: Overall, 9153 YGJ decoction and 576 ALF-related targets were obtained, and 469 YGJ decoction and ALF cross targets were obtained. Of these, 20 key targets, including AKT1, estrogen receptor 1 (ESR1), catalase (CAT), interleukin-1β (IL-1β) and discs large homolog 4 (DLG4), were selected from the PPI network. GO function enrichment analysis revealed that these targets were primarily associated with regulating system processes, cell body, oxidoreductase activity and other processes. An enrichment analysis using the KEGG pathway database revealed that the treatment of ALF using the YGJ decoction was primarily associated with the AGE-RAGE, cGMP-PKG and HIF-1 signalling pathways. Molecular docking indicated that quercetin, stigmasterol and β- sitosterol as well as AKT1, ESR1, CAT, IL-1β and DLG4 exhibited good binding affinity. In vitro experiments revealed that the YGJ decoction significantly reversed lipopolysaccharide (LPS)-induced apoptosis (P < 0.01), inhibited LPS-induced increase in IL-1β and ESR1 levels and upregulated LPS-induced decrease in AKT1, CAT and DLG4 levels (P < 0.05).

Conclusions: The YGJ decoction alleviates ALF by regulating multiple targets and its action mechanism may be associated with ameliorating oxidative stress and inflammation.

简介：通过网络药理学、分子对接和细胞实验确定一贯煎汤治疗急性肝衰竭（ALF）的潜在靶点和作用机制。

方法：利用中医药理学系统（TCMSP）和BATMAN-TCM数据库寻找YGJ汤剂靶标。Genecard 数据库用于预测与 ALF 相关的目标。Cytascape 软件用于可视化和选择常见目标，同时建立蛋白质-蛋白质相互作用 (PPI) 网络。通过基因本体论（GO）和京都基因和基因组百科全书（KEGG）途径富集分析筛选核心靶标并评估假定功能。使用AutodockTools、PyMoL和Discovery Studio软件进行分子对接，以验证YGJ汤剂与选定目标之间的相关性。采用MTT法测定YGJ汤对Huh-7人肝癌细胞活力的影响。通过蛋白质印迹和定量实时聚合酶链反应分析确定YGJ汤剂对Huh-7细胞中推定靶标表达的影响。

结果：总体获得了9153个YGJ汤和576个ALF相关目标，469个YGJ汤和ALF交叉目标。其中，从 PPI 网络中选择了 20 个关键靶标，包括 AKT1、雌激素受体 1 (ESR1)、过氧化氢酶 (CAT)、白介素-1β (IL-1β) 和椎间盘大同源物 4 (DLG4)。GO功能富集分析显示，这些靶点主要与调节系统过程、细胞体、氧化还原酶活性等过程相关。KEGG通路数据库富集分析显示，益气健汤治疗ALF主要与AGE-RAGE、cGMP-PKG和HIF-1信号通路相关。分子对接表明槲皮素、豆甾醇和β-谷甾醇以及AKT1、ESR1、CAT、IL-1β和DLG4表现出良好的结合亲和力。体外实验结果表明，益气健汤可显着逆转脂多糖（LPS）诱导的细胞凋亡（P < 0.01），抑制 LPS 诱导的 IL-1β 和 ESR1 水平升高，上调 LPS 诱导的 AKT1、CAT 和 DLG4 水平下降（P < 0.05）。

结论:益气健汤通过调节多靶点减轻ALF,其作用机制可能与改善氧化应激和炎症有关。