目的 利用生物信息学、网络药理学、分子对接、动力学模拟方法探讨冬虫夏草治疗非小细胞肺癌的关键靶点及其分子机制。方法 利用TCMSP数据库检索冬虫夏草的活性成分和靶点，并利用GEO数据库获取与非小细胞肺癌相关的数据集，使用R语言软件进行差异分析，获取差异表达基因（DEGs）。通过WGCNA分析确定与肿瘤发生密切相关的基因模块，并取交集获取非小细胞肺癌的枢纽基因，将这些枢纽基因与药物靶点进行映射，利用Cytoscape软件构建“活性成分–疾病枢纽靶点”网络图。使用String数据库和Cytoscape构建靶标蛋白相互作用（PPI）网络，筛选出关键靶点。利用Enrihr在线工具进行基因本体（GO）和京都基因与基因组百科全书（KEGG）富集分析。利用CB-Dock2平台和Discovery Studio 2021Client软件对关键成分和靶点进行分子对接验证和可视化。最后利用Gromacs v2022.03软件对分子对接所得的复合物进行100 ns分子动力学模拟分析。结果 共筛选得到冬虫夏草活性成分10个，自身靶点276个和作用于非小细胞肺癌的靶点9个，通过对虫草甾醇和花生四烯酸2个核心成分富集分析提示，主要涉及氧化物酶体增殖物激活受体（PPAR）信号通路、神经活性配体与受体的相互作用、脂肪细胞的脂肪分解调节、腺苷酸活化蛋白激酶（AMPK）信号通路、叉头框转录因子（FoxO）信号通路等。分子对接结果显示，虫草甾醇和花生四烯酸与过氧化物酶体增殖物激活受体γ（PPARG）、1型血管紧张素II受体（AGTR1）和内皮PAS结构域蛋白1（EPAS1）这3个核心靶点具有良好的结合能力，动力学模拟进一步验证虫草甾醇与PPARG、AGTR1复合物结合稳定。结论 冬虫夏草可能通过调控巨噬细胞极化、神经活性配体与受体、脂质代谢、氧化应激、免疫调节等过程，多种协同作用发挥抗非小细胞肺癌作用。

Objective To explore the key targets of Cordyceps in treatment of non-small cell lung cancer, and its molecular mechanisms using bioinformatics, network pharmacology, molecular docking, and kinetic simulation. Methods The TCMSP database was used to retrieve the active ingredients and targets of Cordyceps, and the GEO database was used to obtain the datasets related to non-small cell lung cancer, and differential expression genes (DEGs) were obtained by differential analysis using R language software. The WGCNA analysis was used to identify the gene modules closely related to tumorigenesis, and the intersection was used to obtain the hub genes of non-small cell lung cancer, which were mapped to the drug targets, and Cytoscape software was used to construct the “active ingredient-disease hub targets” network diagram. The target protein interactions (PPI) network was constructed using the String database and Cytoscape, and the key targets were screened out. GO and KEGG enrichment analysis was performed using Enrihr online tools. Molecular docking verification and visualization of key components and targets were performed using CB-Dock2 platform and Discovery Studio 2021Client software. Finally, 100 ns molecular dynamics simulations were performed on the complexes obtained from molecular docking using Gromacs v2022.03 software. Results A total of 10 active components of Cordyceps were screened, 276 self-targets and 9 non-small cell lung cancer targets. The enrichment analysis of the two core components, cordycepsterol and arachidonic acid, suggested that they were mainly involved in the PPAR signaling pathway, the interaction between the neuroactive ligands and the receptor, the regulation of lipolysis of adipocytes, the AMPK signaling pathway, and the FoxO signaling pathway. The molecular docking results showed that cordycepsterol and arachidonic acid had good binding ability with the three core targets of PPARG, AGTR1, and EPAS1, and the kinetic simulation further verified that the cordycepsterol binds stably to the PPARG and AGTR1 complexes. Conclusion Cordyceps may exert anti- non-small cell lung cancer effects through multiple synergistic effects by modulating macrophage polarization, neuroactive ligands and receptors, lipid metabolism, oxidative stress, immunomodulation and other processes.

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广西青年岐黄学者培养项目(GXQH202422)