青蒿琥酯（ART）是一种从青蒿中提取的天然化合物，在治疗多种肿瘤方面显示出良好的临床潜力，但其确切机制尚不清楚。脉络膜黑色素瘤（CM）是成人主要的恶性眼肿瘤，以其恶性程度高、预后差而闻名，目前的治疗效果有限。本研究采用网络药理学、分子对接和实验验证相结合的方式探讨ART的抗CM作用和机制。在PubChem、Swiss Target Prediction和传统中药系统药理学（TCMSP）数据库分析平台数据库中筛选ART的潜在靶点，而与 CM 预后相关的靶基因选自在线人类孟德尔遗传 (OMIM)、GeneCards 和 DisGeNET 数据库。这两组数据集的交集产生了参与 CM 的 ART 靶基因。对交叉靶标进行蛋白质-蛋白质相互作用（PPI）网络分析，以及基因本体论（GO）和京都基因和基因组百科全书（KEGG）分析，以确定核心靶标和关键途径。进行分子对接方法来预测 ART 与核心靶标之间的结合相互作用。通过 CCK8、集落形成、Transwell 以及流式细胞术检测细胞凋亡、细胞周期、活性氧 (ROS) 来评估 ART 对 CM 的影响。进行蛋白质印迹 (WB) 测定以研究 ART 对与 CM 相关的关键蛋白质和途径的影响。最后，进行体内实验，进一步验证ART对裸鼠皮下肿瘤的作用。研究表明，通过网络药理学方法确定了ART治疗CM的关键途径和核心靶点。分子对接结果验证了ART与这些核心靶点之间的强结合亲和力。分析和预测结果表明，ART主要通过细胞凋亡等多种肿瘤相关途径对CM发挥作用。体外实验证实ART显着抑制CM细胞的增殖和迁移。这是通过激活p53信号通路促进细胞凋亡、通过抑制PI3K/AKT/mTOR信号通路引起细胞周期停滞在G0/G1期以及通过激活NRF2/HO-1信号通路增加细胞内ROS水平来实现的。途径。此外，体内实验进一步验证了ART对CM显着的增殖抑制作用。本研究通过网络药理学结合分子对接和体内外实验进行了初步探索，证实了ART具有潜在的抗CM作用。癌症通过促进细胞凋亡、诱导细胞周期停滞和增加细胞内 ROS 水平对 CM 产生影响。这些研究结果表明，ART 对 CM 具有显着的治疗潜力。版权所有 © 2024 Ma、Liu、Zhang、Yi、Sun、Gao、Zhao、Wang、Lei 和 Luo。

Artesunate (ART), a natural compound derived from Artemisia annua, has shown promising clinical potentials in the treatment of various tumors, but the exact mechanism is unclear. Choroidal melanoma (CM) is a major malignant ocular tumor in adults, known for its significant malignancy and poor prognosis, with limited efficacy in current treatments. This study explored the anti-CM effects and mechanisms of ART using a combination of network pharmacology, molecular docking and experimental validation.Potential targets of ART were screened in PubChem, Swiss Target Prediction and Traditional Chinese Medicine Systems Pharmacology (TCMSP) Database Analysis Platform databases, while target genes related to CM prognosis were selected from Online Mendelian Inheritance in Man (OMIM), GeneCards and DisGeNET databases. The intersection of these two groups of datasets yielded the target genes of ART involved in CM. Protein-protein interaction (PPI) network analysis of the intersecting targets, as well as Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses, were conducted to identify core targets and critical pathways. Molecular docking methods were performed to predict the binding interactions between ART and core targets. The effects of ART on CM were evaluated through CCK8, colony formation, transwell, as well as flow cytometry assays to detect apoptosis, cell cycle, reactive oxygen species (ROS). Western blot (WB) assays were conducted to investigate the impact of ART on key proteins and pathways associated with CM. Finally, in vivo assays were conducted to further validate the effects of ART on subcutaneous tumors in nude mice.Research has shown that key pathways and core targets for ART in treating CM were identified through a network pharmacology approach. Molecular docking results verified the strong binding affinity between ART and these core targets. The analysis and predicted results indicated that ART primarily exerted its effects on CM through various tumor-related pathways like apoptosis. The assays in vitro confirmed that ART significantly inhibited the proliferation and migration of CM cells. This was achieved by promoting apoptosis through activation of the p53 signaling pathway, causing cell cycle arrest at the G0/G1 phase by inhibiting the PI3K/AKT/mTOR signaling pathway and increasing the intracellular level of ROS by activating the NRF2/HO-1 signaling pathway. Additionally, the assays in vivo further validated the significant proliferation-inhibitory effect of ART on CM.This study, making the initial exploration, illustrated through network pharmacology combined with molecular docking and in vitro/in vivo assays, confirmed that ART exerted potential anti-cancer effects on CM by promoting apoptosis, inducing cell cycle arrest and increasing intracellular levels of ROS. These findings suggested that ART held significant therapeutic potential for CM.Copyright © 2024 Ma, Liu, Zhang, Yi, Sun, Gao, Zhao, Wang, Lei and Luo.