目的：芦荟大黄素（AE）是从天然植物大黄的根茎中提取的蒽醌类化合物。最初，研究表明AE具有抗炎作用。进一步的研究揭示了它对多种癌症的抗肿瘤活性。然而，这些特性背后的机制仍不清楚。本研究基于网络药理学和分子对接，探讨AE治疗肝细胞癌（HCC）的分子机制，并通过体外实验评价其治疗效果。方法：CTD、Pharmmapper、SuperPred和TargetNet是获取潜在药物相关靶点的数据库。 DisGenet、GeneCards、OMIM 和 TTD 用于识别潜在的疾病相关目标。通过维恩图获得药物和疾病的交叉基因。 Bioinformatics网站对交叉基因进行了基因本体论（GO）和京都基因与基因组百科全书（KEGG）富集分析。将交叉基因引入STRING中构建蛋白质-蛋白质相互作用网络，同时使用Cytoscape3.9.1软件对核心靶标进行可视化和分析。利用AutoDock4.2.6实现药物与核心靶点的分子对接。体外实验研究了AE的治疗效果和相关机制。结果：获得了 63 个重叠基因，GO 分析通过这 63 个交叉基因生成了 3,646 个条目。 KEGG分析主要涉及细胞凋亡、癌症蛋白聚糖、TNF信号通路、TP53信号通路、PI3K-AKT信号通路等。AKT1、EGFR、ESR1、TP53、SRC因其与芦荟之间的结合能而被确定为核心靶点。 -大黄素小于-5 kcal/Mol。进一步揭示了与核心靶标相关的mRNA和蛋白表达、预后、突变状态和免疫浸润。 AKT1和EGFR的参与以及PI3K-AKT信号通路的关键靶点表明该信号通路在AE治疗HCC中的重要性。 Cell Counting Kit-8 检测和流式分析的结果证明了 AE 的治疗效果。 EGFR、PI3KR1、AKT1 和 BCL2 mRNA 表达的下调以及 PI3KR1、AKT、p-AKT 蛋白表达的下调证实了我们的假设。结论：基于网络药理学和分子对接，我们的研究初步表明AE通过调节多种信号通路对HCC发挥治疗作用。各种分析证实了 AE 通过 PI3K-AKT 信号通路对 HCC 具有抗增殖活性和促凋亡作用。该研究通过一种新的方法揭示了AE治疗HCC的治疗机制，为AE的临床应用提供了理论基础。版权所有©2023 Zhu，He，Wang，Duan，Rong，Wu，Ding，Mi，Ge，杨、于。

Objective: Aloe-emodin (AE) is an anthraquinone compound extracted from the rhizome of the natural plant rhubarb. Initially, it was shown that AE exerts an anti-inflammatory effect. Further studies revealed its antitumor activity against various types of cancer. However, the mechanisms underlying these properties remain unclear. Based on network pharmacology and molecular docking, this study investigated the molecular mechanism of AE in the treatment of hepatocellular carcinoma (HCC), and evaluated its therapeutic effect through in vitro experiments. Methods: CTD, Pharmmapper, SuperPred and TargetNet were the databases to obtain potential drug-related targets. DisGenet, GeneCards, OMIM and TTD were used to identify potential disease-related targets. Intersection genes for drugs and diseases were obtained through the Venn diagram. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses of intersecting genes were conducted by the website of Bioinformatics. Intersection genes were introduced into STRING to construct a protein-protein interaction network, while the Cytoscape3.9.1 software was used to visualize and analyze the core targets. AutoDock4.2.6 was utilized to achieve molecular docking between drug and core targets. In vitro experiments investigated the therapeutic effects and related mechanisms of AE. Results: 63 overlapped genes were obtained and GO analysis generated 3,646 entries by these 63 intersecting genes. KEGG analysis mainly involved apoptosis, proteoglycans in cancer, TNF signaling pathway, TP53 signaling pathway, PI3K-AKT signaling pathway, etc. AKT1, EGFR, ESR1, TP53, and SRC have been identified as core targets because the binding energies of them between aloe-emodin were less than -5 kcal/Mol.The mRNA and protein expression, prognosis, mutation status, and immune infiltration related to core targets were further revealed. The involvement of AKT1 and EGFR, as well as the key target of the PI3K-AKT signaling pathway, indicated the importance of this signaling pathway in the treatment of HCC using AE. The results of the Cell Counting Kit-8 assay and flow analysis demonstrated the therapeutic effect of AE. The downregulation of EGFR, PI3KR1, AKT1, and BCL2 in mRNA expression and PI3KR1, AKT,p-AKT in protein expression confirmed our hypothesis. Conclusion: Based on network pharmacology and molecular docking, our study initially showed that AE exerted a therapeutic effect on HCC by modulating multiple signaling pathways. Various analyses confirmed the antiproliferative activity and pro-apoptotic effect of AE on HCC through the PI3K-AKT signaling pathway. This study revealed the therapeutic mechanism of AE in the treatment of HCC through a novel approach, providing a theoretical basis for the clinical application of AE.Copyright © 2023 Zhu, He, Wang, Duan, Rong, Wu, Ding, Mi, Ge, Yang and Yu.