目的 结合网络药理学与体外实验验证探讨丹皮酚抑制血管内皮细胞间质转化（EndMT）的关键靶点及作用机制。方法 通过PharmMapper、HERB、SymMap、ChEMBL、SwissTargetPredicition数据库获取丹皮酚的作用靶点；从GeneCards数据库中检索与EndMT相关的靶标，将两者进行交集分析与可视化；将交集靶点基因上传至STRING数据库联用Cytoscape 3.8.2软件构建蛋白相互作用（PPI）网络并筛选核心靶点基因，并进行基因本体论（GO）功能富集和京都基因与基因组百科全书（KEGG）信号通路富集分析。采用Autodock Vina软件对丹皮酚与核心靶点基因进行分子对接验证。体外实验将HUVECs细胞分为空白组、模型[转化生长因子β1（TGF-β1）]组、丹皮酚低、中、高剂量组（30、60、120 μmol/L），通过蛋白质印迹法（Western blotting）筛选TGF-β1诱导EndMT最佳浓度并通过酶联免疫吸附测定（ELISA）检测基质金属蛋白酶2（MMP-2）、基质金属蛋白酶9（MMP-9）的蛋白表达量。结果 共筛选出丹皮酚作用靶点414个，EndMT相关靶点基因241个，取交集后得到27个丹皮酚治疗EndMT的潜在作用靶点。核心靶点与京都基因与基因组百科全书（KEGG）通路分析结果显示，动脉粥样硬化信号通路是丹皮酚涉及EndMT的主要通路。分子对接结果显示，丹皮酚与MMP-2与MMP-9的结合能力最强。ELISA实验证明，丹皮酚可以减少MMP-2、MMP-9蛋白的表达（P＜0.01）。结论 丹皮酚抑制EndMT的关键靶点是MMP-2与MMP-9，为进一步研究丹皮酚抗动脉粥样硬化的作用机制提供依据。

Objective To explore the key targets and mechanism of paeonol inhibiting vascular endothelial interstitial transformation (EndMT) by network pharmacology and in vitro experiments. Methods PharmMapper, HERB, SymMap, ChEMBL, and SwissTargetPredicition were used to predict the target of paeonol. GeneCards was used to search for targets related to EndMT. Paeonol and EndMT targets were used for intersection analysis visualization. The overlapped genes were mapped to the STRING database, and protein-protein interactions (PPI) were built using the STRING database and Cytoscape 3.8.2, and the core target genes were screened. The gene ontology analysis (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were then performed. AutoDock Vina was used to conduct the molecular docking between paeonol and the core target genes. In vitro experiment, HUVECs cells were divided into blank group, model [transforming growth factor β1 (TGF-β1)] group, paeonol low-dose, medium-dose and high-dose groups (30, 60, 120 μmol/L). The optimal concentration of EndMT induced by TGF-β1 was screened by Western blotting, and the protein expressions of matrix metalloproteinase 2 (MMP-2) and matrix metalloproteinase 9 (MMP-9) were detected by enzyme-linked immunosorbent assay (ELISA). Results A total of 414 functional targets of paeonol and 241 EndMT-related targets were screened by network pharmacology analysis. By taking the intersection, a total of 27 potential targets of the regulatory effect of paeonol on EndMT were found. The core target and KEGG pathway analyses showed that the atherosclerotic signaling pathway was the key pathway for paeonol in treatment of EndMT. Molecular docking results showed that paeonol had a better binding ability with MMP-2 and MMP-9. ELISA results indicated that paeonol could reduce the expression of MMP-2 and MMP-9 (P < 0.01). Conclusion The key targets of paeonol inhibiting EndMT are MMP-2 and MMP-9, which provides a basis for further research on the mechanism of paeonol anti-atherosclerosis.

R966

国家自然科学基金资助项目（82174014）