目的 整合生物信息学、网络药理学及分子对接技术预测麦冬皂苷D（OP-D）治疗特发性肺纤维化（IPF） 的作用机制，并进行细胞实验验证。方法 （1）利用 R软件的 Limma包分析 GEO数据库中 IPF差异表达基因数 据，筛选 IPF 疾病靶点；采用 PharmMapper和 SwissTargetPrediction 数据库预测 OP-D作用靶点；对筛选得到的 OP-D作用靶点与IPF疾病靶点取交集，得到OP-D治疗IPF的潜在作用靶点。对潜在作用靶点进行GO功能及 KEGG 通路富集分析；采用 STRING 数据库和 Cytoscape 软件构建潜在作用靶点的蛋白互作（PPI）网络，并筛选 OP-D抗IPF的核心靶点。使用Autodock Vina软件对OP-D与核心靶点进行分子对接验证。（2）采用TGF-β（10 μg·L-1）诱 导人胚肺成纤维细胞（HFL-1），并用不同浓度OP-D进行干预。采用CCK-8法、EdU法检测细胞增殖情况；Transwell 检测细胞迁移能力；TUNEL染色法检测细胞凋亡情况；Western Blot法检测细胞中相关蛋白的表达水平。结果 （1）筛 选得到IPF疾病靶点2 040个，OP-D作用靶点408个，取交集得到30个OP-D治疗IPF的潜在作用靶点。OP-D 抗 IPF的生物过程主要在胶原蛋白分解代谢过程、胶原蛋白代谢过程、细胞外基质分解和对异生物刺激的反应 方面；主要涉及通路包括肥厚型心肌病信号通路、类风湿性关节炎信号通路、松弛素信号通路、PPAR信号通 路和IL-17信号通路等。进一步筛选得到IGF1、MMP1、MMP2、MMP3、MMP7、ACE、CCL5等OP-D抗IPF的 核心靶点，分子对接显示 OP-D 与核心靶点均有较好的结合活性。（2）OP-D 能浓度依赖性抑制 HFL-1 细胞的 活力，选择1、2、4 μmol·L-1 OP-D进行后续实验。与正常对照组比较，TGF-β组细胞的EdU阳性率及迁移细 胞数显著升高（P < 0.01）；TUNEL 阳性率及 Bax 蛋白表达水平显著下降（P < 0.01），Bcl-2、FN-1、α-SMA、 Collagen Ⅰ、IGF1、MMP1、MMP2、MMP3、MMP7蛋白表达水平显著升高（P < 0.05，P < 0.01）。与 TGF-β 组 比较，1、2、4 μmol·L-1 OP-D组的细胞 EdU阳性率与迁移细胞数显著降低（P < 0.05，P < 0.01），TUNEL阳性 率及Bax蛋白表达水平显著升高（P < 0.05，P < 0.01），Bcl-2、IGF1、MMP1、MMP2、MMP3、MMP7蛋白表达水平 显著下降（P < 0.05，P < 0.01）；4 μmol·L-1OP-D 组细胞的 FN-1、Collagen Ⅰ蛋白表达水平显著下降（P < 0.05， P < 0.01）；2、4 μmol·L-1 OP-D组细胞的α-SMA蛋白表达水平显著下降（P < 0.05，P < 0.01）。结论 OP-D可 能通过抑制 IGF1表达，降低 MMPs表达水平，进而抑制 HFL-1细胞增殖、迁移、凋亡抵抗和纤维化，从而发 挥抗IPF的作用。

Objective To integrate bioinformatics，network pharmacology and molecular docking techniques to predict the mechanism of ophiopogonin D（OP-D）in the treatment of idiopathic pulmonary fibrosis（IPF）and to verify it by cell experiments. Methods（1）The Limma package for R software was used to analyze the IPF gene expression data in the GEO database and to screen the IPF disease targets. PharmMapper and SwissTargetPrediction databases were used to predict potential OP-D targets，and targets of OP-D for the treatment of IPF were screened by taking the intersection of OP-D targets and IPF targets. Gene Ontology（GO）enrichment analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG）pathway analysis were performed on the target. The STRING database and Cytoscape software were used to construct the protein-protein interaction（PPI）network of potential targets，and the core targets of OP-D for anti-IPF were screened. Autodock Vina software was used to perform molecular docking between OP-D and core targets. （2） TGF-β（10 μg·L-1）was used to induce human embryonic lung fibroblasts，which were treated with OP-D at different concentration. CCK-8 and EdU methods were applied to detect cell proliferation. Transwell system was employed to determine cell migration. TUNEL staining was used to measure cell apoptosis. Western Blot method was applied to detect the expression levels of related proteins in cells. Results（1）A total of 2 040 IPF disease targets and 408 OP-D targets were screened. The intersection of the two were taken to obtain 30 potential targets of OP-D for the treatment of IPF. GO and KEGG analysis showed that the main biological processes of OP-D for the treatment of IPF involved in collagen catabolism， collagen metabolism， extracellular matrix decomposition， and the stimulus of xenobiotics， and the enrichment pathways were mainly involved in hypertrophic cardiomyopathy signaling pathway， rheumatoid arthritis signaling pathway， relaxin signaling pathway， PPAR signaling pathway and IL-17 signaling pathway. The core targets of OP-D for anti-IPF were IGF-1， MMP1， MMP2， MMP3， MMP7， ACE and CCL5. Molecular docking indicated that OP-D had good binding activity with core targets.（2）OP-D can concentrationdependently inhibit the viability of HFL-1 cells， and 1， 2， and 4 μmol · L-1 OP-D were selected for subsequent experiments. Compared with the normal control group，the EdU positive rate and number of cell migration in TGF- β group were significantly increased（P < 0.01）. TUNEL positive rate and Bax protein expression level were significantly decreased（P < 0.01）. The protein expression levels of Bcl-2，FN-1，α-SMA，CollagenⅠ，IGF1，MMP1，MMP2， MMP3 and MMP7 were significantly increased（P < 0.05，P < 0.01）. Compared with TGF-β group，the EdU positive rate and number of cell migration in OP-D group at different concentration were significantly decreased（P < 0.05，P < 0.01）. TUNEL positive rate and Bax protein expression level were significantly increased（P < 0.05，P < 0.01）. The protein expression levels of Bcl-2，IGF1，MMP1，MMP2，MMP3 and MMP7 were significantly decreased（P < 0.05，P < 0.01）. The protein expression levels of FN-1 and Collagen Ⅰ in 4 μmol·L-1 OP-D group were significantly decreased （P < 0.05，P < 0.01），while α-SMA protein expression was significantly decreased in 2 and 4 μmol·L-1 OP-D groups （P < 0.05，P < 0.01）. Conclusion OP-D may play an anti-IPF role by inhibiting IGF1 expression and down-regulating thelevel ofMMPs，therebyinhibitingtheproliferation，migration，apoptosisresistance andfibrosis ofHFL-1 cells.

R285.5；R857.3

国家自然科学基金项目（82004306）；湖南省中医药科研计划项目（B2023017）；湖南省自然科学基金项目（2024JJ5237）