目的 基于转录组学和网络药理学探讨肉豆蔻挥发油（volatile oil from Myristica fragrans，VOMF）干预低氧性肺动脉高压（hypoxic pulmonary artery hypertension，HPAH）的作用机制。方法 通过SwissTargetPrediction数据库获取VOMF活性成分和药物潜在靶点，应用GEO数据库获取HPAH疾病相关靶点，Cytoscape软件构建“成分-靶点-疾病”网络；运用Pymol和MOE软件对VOMF关键活性成分与作用靶点进行分子对接。采用雄性SD大鼠制备肺动脉平滑肌细胞（pulmonary artery smooth muscle cells，PASMCs），低氧处理并给予VOMF干预后，进行转录组学分析获取差异表达基因，采用Western blotting验证VOMF对细胞周期信号通路相关蛋白表达的影响。结果 网络药理学分析共得到VOMF潜在作用靶点512个，HPAH疾病相关差异靶点2 912个，交集基因80个；转录组学分析共得到40个共同差异表达基因。以P＜0.05为筛选条件将转录组学和京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）通路富集分析结果进行交集，得到交集通路8条，其中与HPAH密切相关的包括细胞周期信号通路、丝裂原活化蛋白激酶信号通路、缺氧诱导因子-1信号通路。结合文献报道和课题组前期研究，选取细胞周期信号通路进行验证。Western blotting结果显示，与对照组比较，模型组细胞周期蛋白依赖性激酶2（cyclin dependent kinase 2，CDK2）、CDK4、细胞周期蛋白D1（cell cycle protein D1，Cyclin D1）、Cyclin A2蛋白表达水平升高（P＜0.01、0.001），p27Kip1蛋白表达水平降低（P＜0.01）；与模型组比较，VOMF组CDK2、CDK4、Cyclin D1、Cyclin A2蛋白表达水平降低（P＜0.05、0.01、0.001），p27Kip1蛋白表达水平升高（P＜0.05、0.01）。结论 从网络药理学和转录组学角度初步阐明了VOMF能够通过干预细胞周期信号通路，进而干预HPAH的发生，可为后续的药理学研究和临床应用提供依据与参考。

Objective To explore the mechanism of volatile oil of Myristica fragrans (VOMF) on hypoxic pulmonary artery hypertension (HPAH) based on transcriptomics and network pharmacology. Methods Active components and potential drug targets of VOMF were obtained through SwissTargetPrediction database, HPAH disease related targets were obtained through GEO database, and “component-target-disease” network was constructed by Cytoscape software; Pymol and MOE software were used to verify the key active components and targets of VOMF by molecular docking. Male SD rats were used to prepare pulmonary artery smooth muscle cells (PASMCs), after given hypoxia treatment and VOMF intervention, transcriptomic analysis was performed to obtain differentially expressed genes, and Western blotting was used to verify the effect of VOMF on expressions of cell cycle signaling pathway related proteins. Results A total of 512 potential targets of VOMF, 2 912 differential targets related to HPAH disease and 80 intersection genes were obtained by network pharmacology analysis. A total of 40 common differentially expressed genes were obtained by transcriptome analysis. The results of transcriptomics and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis were intersected with P < 0.05 as the screening condition, and eight intersection pathways were obtained. Among them, the cell cycle signaling pathway, mitogen-activated protein kinase signaling pathway and hypoxia inducible factor-1 signaling pathway were closely related to HPAH. Based on literature research and previous research by the research group, cell cycle signaling pathway was selected for validation. The results of Western blotting experiments showed that compared with control group, cyclin dependent kinase 2 (CDK2), CDK4, cell cycle protein D1 (Cyclin D1), Cyclin A2 protein expression levels in model group were increased (P < 0.01, 0.001), p27Kip1 protein expression level was decreased (P < 0.01); Compared with model group, CDK2, CDK4, Cyclin D1, Cyclin A2 protein expression levels in VOMF group were decreased (P < 0.05, 0.01, 0.001), p27Kip1 protein expression level was increased (P < 0.05, 0.01). Conclusion From the perspectives of network pharmacology and transcriptomics, it has been preliminarily elucidated that VOMF can intervene in the occurrence of HPAH by intervening in the cell cycle signaling pathway, providing a basis and reference for subsequent pharmacological research and clinical applications.

R285.5

青海省科技厅自然科学基金项目面上项目（2021-ZJ-907）；国家自然科学基金资助项目（32060088）；国家自然科学基金面上项目（82374148）；国家自然科学基金资助项目（82060786）