DataSheet1_RETRACTED: Mechanism of action of Asparagus officinalis extract against multiple myeloma using bioinformatics tools, in silico and in vitro study.docx

Introduction: Asparagus (Asparagus officinalis) is a perennial flowering plant species. Its main components have tumor-prevention, immune system-enhancement, and anti-inflammation effects. Network pharmacology is a powerful approach that is being applied increasingly to research of herbal medicines. Herb identification, study of compound targets, network construction, and network analysis have been used to elucidate how herbal medicines work. However, the interaction of bioactive substances from asparagus with the targets involved in multiple myeloma (MM) has not been elucidated. We explored the mechanism of action of asparagus in MM through network pharmacology and experimental verification.
Methods: The active ingredients and corresponding targets of asparagus were acquired from the Traditional Chinese Medicine System Pharmacology database, followed by identification of MM-related target genes using GeneCards and Online Mendelian Inheritance in Man databases, which were matched with the potential targets of asparagus. Potential targets were identified and a target network of traditional Chinese medicine was constructed. The STRING database and Cytoscape were utilized to create protein–protein interaction (PPI) networks and further screening of core targets.
Results: The intersection of target genes and core target genes of the phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) pathway was enriched, the top-five core target genes were selected, and the binding affinity of corresponding compounds to the top-five core targets was analyzed using molecular docking. Network pharmacology identified nine active components of asparagus from databases based on oral bioavailability and drug similarity, and predicted 157 potential targets related to asparagus. Enrichment analyses showed that “steroid receptor activity” and the “PI3K/AKT signaling pathway” were the most enriched biological process and signaling pathway, respectively. According to the top-10 core genes and targets of the PPI pathway, AKT1, interleukin (IL)-6, vascular endothelial growth factor (VEGF)A, MYC, and epidermal growth factor receptor (EGFR) were selected for molecular docking. The latter showed that five core targets of the PI3K/AKT signaling pathway could bind to quercetin, among which EGFR, IL-6, and MYC showed strong docking, and the diosgenin ligand could bind to VEGFA. Cell experiments showed that asparagus, through the PI3K/AKT/NF-κB pathway, inhibited the proliferation and migration of MM cells, and caused retardation and apoptosis of MM cells in the G0/G1 phase.
Discussion: In this study, the anti-cancer activity of asparagus against MM was demonstrated using network pharmacology, and potential pharmacological mechanisms were inferred using in vitro experimental data.

引言：石刁柏（Asparagus officinalis）是一种多年生显花植物。其主要成分具有抗肿瘤、增强免疫功能及抗炎作用。网络药理学是一种日益广泛应用于中药研究的有力研究方法。中药研究常通过药物识别、化合物靶点研究、网络构建及网络分析来阐明其作用机制，但目前尚未明确石刁柏中的生物活性物质与多发性骨髓瘤（multiple myeloma, MM）相关靶点的相互作用。本研究借助网络药理学结合实验验证，探讨石刁柏治疗多发性骨髓瘤的作用机制。
方法：从中药系统药理学数据库（Traditional Chinese Medicine System Pharmacology database）中获取石刁柏的活性成分及其对应靶点；通过GeneCards数据库与在线人类孟德尔遗传（Online Mendelian Inheritance in Man）数据库筛选多发性骨髓瘤相关靶基因，并与石刁柏潜在靶点进行匹配。筛选潜在靶点并构建中药靶点网络。利用STRING数据库与Cytoscape软件构建蛋白质相互作用（protein–protein interaction, PPI）网络，进一步筛选核心靶点。
结果：本研究富集得到靶基因交集及磷脂酰肌醇3-激酶/蛋白激酶B（phosphoinositide 3-kinase/protein kinase B, PI3K/AKT）通路核心靶基因，选取排名前五的核心靶点，通过分子对接（molecular docking）分析对应化合物与前五核心靶点的结合活性。基于口服生物利用度与药物相似性，网络药理学从数据库中筛选得到石刁柏的9种活性成分，并预测得到157个石刁柏相关潜在靶点。富集分析显示，“类固醇受体活性（steroid receptor activity）”与“PI3K/AKT信号通路（phosphoinositide 3-kinase/protein kinase B signaling pathway）”分别为最显著富集的生物过程与信号通路。根据PPI通路排名前十的核心基因与靶点，选取AKT1、白细胞介素（interleukin, IL)-6、血管内皮生长因子（vascular endothelial growth factor, VEGF)A、MYC及表皮生长因子受体（epidermal growth factor receptor, EGFR）进行分子对接，结果显示PI3K/AKT信号通路的5个核心靶点可与槲皮素结合，其中EGFR、IL-6及MYC的对接结合活性较强；薯蓣皂苷元配体可与VEGFA结合。细胞实验证实，石刁柏可通过PI3K/AKT/NF-κB通路抑制多发性骨髓瘤细胞的增殖与迁移，并使细胞阻滞于G0/G1期且诱导其凋亡。
讨论：本研究通过网络药理学验证了石刁柏抗多发性骨髓瘤的活性，并结合体外（in vitro）实验数据推断了其潜在药理机制。