目的 利用生物信息學(xué)、網(wǎng)絡(luò)藥理學(xué)、分子對接、動力學(xué)模擬方法探討冬蟲夏草治療非小細胞肺癌的關(guān)鍵靶點及其分子機制。方法 利用TCMSP數(shù)據(jù)庫檢索冬蟲夏草的活性成分和靶點，并利用GEO數(shù)據(jù)庫獲取與非小細胞肺癌相關(guān)的數(shù)據(jù)集，使用R語言軟件進行差異分析，獲取差異表達基因（DEGs）。通過WGCNA分析確定與腫瘤發(fā)生密切相關(guān)的基因模塊，并取交集獲取非小細胞肺癌的樞紐基因，將這些樞紐基因與藥物靶點進行映射，利用Cytoscape軟件構(gòu)建“活性成分–疾病樞紐靶點”網(wǎng)絡(luò)圖。使用String數(shù)據(jù)庫和Cytoscape構(gòu)建靶標(biāo)蛋白相互作用（PPI）網(wǎng)絡(luò)，篩選出關(guān)鍵靶點。利用Enrihr在線工具進行基因本體（GO）和京都基因與基因組百科全書（KEGG）富集分析。利用CB-Dock2平臺和Discovery Studio 2021Client軟件對關(guān)鍵成分和靶點進行分子對接驗證和可視化。最后利用Gromacs v2022.03軟件對分子對接所得的復(fù)合物進行100 ns分子動力學(xué)模擬分析。結(jié)果 共篩選得到冬蟲夏草活性成分10個，自身靶點276個和作用于非小細胞肺癌的靶點9個，通過對蟲草甾醇和花生四烯酸2個核心成分富集分析提示，主要涉及氧化物酶體增殖物激活受體（PPAR）信號通路、神經(jīng)活性配體與受體的相互作用、脂肪細胞的脂肪分解調(diào)節(jié)、腺苷酸活化蛋白激酶（AMPK）信號通路、叉頭框轉(zhuǎn)錄因子（FoxO）信號通路等。分子對接結(jié)果顯示，蟲草甾醇和花生四烯酸與過氧化物酶體增殖物激活受體γ（PPARG）、1型血管緊張素II受體（AGTR1）和內(nèi)皮PAS結(jié)構(gòu)域蛋白1（EPAS1）這3個核心靶點具有良好的結(jié)合能力，動力學(xué)模擬進一步驗證蟲草甾醇與PPARG、AGTR1復(fù)合物結(jié)合穩(wěn)定。結(jié)論 冬蟲夏草可能通過調(diào)控巨噬細胞極化、神經(jīng)活性配體與受體、脂質(zhì)代謝、氧化應(yīng)激、免疫調(diào)節(jié)等過程，多種協(xié)同作用發(fā)揮抗非小細胞肺癌作用。

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.

R285

廣西青年岐黃學(xué)者培養(yǎng)項目(GXQH202422)