DiDang Decoction (DDD)是中国传统经典验方，用于治疗动脉粥样硬化和高脂血症。然而，DDD的作用机制仍不清楚。采用网络药理学和体外实验验证DDD在动脉粥样硬化和高脂血症中的作用机制。从传统中药系统药理数据库和文献挖掘中获取DDD的化学成分，从Gencards、OMIM和DisGeNET数据库中获取动脉粥样硬化和高脂血症的疾病靶点。将交集基因导入STRING数据库构建蛋白质相互作用（PPI）网络，使用DAVID数据库进行基因本体论（GO）和基因组百科全书（KEGG）通路富集分析。结合KEGG通路分析的结果，选择HIF-1信号通路进行进一步的体外实验。结果显示，网络药理学预测DDD治疗动脉粥样硬化和高脂血症的相关靶点有112个，前10个相关通路是：脂肪和动脉粥样硬化、AGE-RAGE信号通路的糖尿病并发症、化学致癌物诱导受体激活、癌症途径、癌症中的蛋白聚糖、剪切应力和动脉粥样硬化、HIF-1信号通路、酒精性肝病、PPAR信号通路和冠状病毒病COVID-19。体外实验表明，DDD有效地减少 FFA 处理的 L02 细胞中的脂质积累；DDD减轻了线粒体损伤并减少了 ROS 含量；DDD抑制了铁死亡和凋亡；DDD上调了 HIF-1α 、谷胱甘肽过氧化物酶4（GPX4）和 Bcl2 蛋白的表达，并下调了 Bax 蛋白的表达。DDD通过多个靶点和通路对动脉粥样硬化和高脂血症发挥治疗作用，促进线粒体功能改善，降低 ROS 含量，抑制铁死亡和凋亡，通过激活 HIF-1 信号通路，为DDD治疗动脉粥样硬化和高脂血症提供了可靠的理论和实验支持。版权所有©2023 Elsevier B.V.。保留所有权利。

DiDang Decoction (DDD) is a traditional classical prescription that has been used to treat atherosclerosis (AS) and hyperlipidemia (HLP) in China. Nevertheless, the underlying mechanism of DDD remains unclear.To validate the mechanism of DDD in AS and HLP based on network pharmacology and in vitro experiments.The chemical components of DDD were obtained from the Traditional Chinese Medicine System Pharmacology Database and Analysis Platform (TCMSP) database and literature mining, and the disease targets of AS and HLP were obtained from the Gencards, OMIM, and DisGeNET databases. The intersection genes were imported into the STRING database to construct protein-protein interaction (PPI) network, and the DAVID database was used for gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Combined with the results of KEGG pathway analysis, the HIF-1 signaling pathway was selected for further in vitro experiments.The results showed that network pharmacology predicted 112 targets related to DDD treatment of AS and HLP, and the top 10 related pathways are: Lipid and atherosclerosis, AGE-RAGE signaling pathway in diabetic complications, Chemical carcinogenesis - receptor activation, Pathways in cancer, Proteoglycans in cancer, Fluid shear stress and atherosclerosis, HIF-1 signaling pathway, Alcoholic liver disease, PPAR signaling pathway, and Coronavirus disease-COVID-19. In vitro experiments showed that DDD effectively reduced lipid accumulation in FFA-treated L02 cells; DDD attenuated mitochondrial damage and reduced ROS content; DDD inhibited ferroptosis and apoptosis; DDD up-regulated the expression of HIF-1α, Glutathione Peroxidase 4(GPX4), and Bcl2 proteins, and down-regulated expression of Bax protein.DDD exerts therapeutic effects on AS and HLP through multiple targets and pathways, and improves mitochondrial function, reduces ROS content, inhibits ferroptosis and apoptosis by activating the HIF-1 signaling pathway, which provides reliable theoretical and experimental support for DDD treatment of AS and HLP.Copyright © 2023 Elsevier B.V. All rights reserved.