【目的】 探讨补肾活血方（出自《伤科大成》）作用于前交叉韧带损伤腱骨愈合的分子机制。【方法】 运用TCMSP数据库 筛选补肾活血方组成中药的活性成分及对应靶点。于GeneCards、OMIM、PharmGkb等疾病靶点数据库检索获得前交叉韧带 损伤腱骨愈合疾病的相关基因。获得活性成分和疾病的共同作用靶点。采用Cytoscape软件构建药物活性成分-疾病靶点关系 网络，String在线软件构建蛋白质相互作用（PPI）网络，并使用Cytoscape的Cyto NCA插件进行网络拓扑学分析，筛选出核心 靶点及对应活性成分。对药物活性成分和疾病的共同靶点进行基因本体论（GO）生物学功能及京都基因与基因组百科全书 （KEGG）通路富集分析 。采用Autodocktools软件将度（Degree）值前5位的活性成分与部分核心靶点进行分子对接并可视化绘 图。【结果】 TCMSP数据库筛选得到补肾活血方组成中药的有效活性成分共121个，潜在靶点243个。于GeneCards（413个）、 OMIM（3个）和PharmGkb（4个）数据库中筛选出前交叉韧带损伤腱骨愈合疾病靶点416个。获得药物活性成分-疾病共同靶点 42个，对应药物活性成分108个。网络拓扑学筛选共得到VEGFA、HIF1A、IL1B、IL6、IL10、TNF、EGF、TP53、CXCL8、 CD44、SPP1、PTGS2、MMP2、MMP3、MMP9和 VCAM1等 16个 PPI网络核心靶点。GO 生物学功能富集于细胞外基质胶原 代谢、活性氧调控、内肽酶及细胞因子活性、整合素结合等生物学功能。KEGG通路富集于动脉粥样硬化、HIF-1等血管相 关信号通路，以及TNF、Toll样受体、NF-κB等炎症相关信号通路，和C型凝集素受体等免疫相关信号通路。基于KEGG富 集结果选取血管生成（VEGFA、HIF1A）及炎症（IL1B、IL6、IL10、TNF、PTGS2）密切相关的核心靶点进行分子对接，结果表 明补肾活血方的关键药物活性成分与前交叉韧带损伤腱骨愈合关键靶点有较好的结合活性。【结论】 补肾活血方可能通过活 性成分槲皮素、木犀草素、山奈酚、β–胡萝卜素、鞣花酸作用于VEGFA、HIF1A、IL1B、IL6、IL10、TNF、PTGS2等靶点 调控血管生成及炎症，促进前交叉韧带损伤腱骨愈合过程。

Objective To investigate the molecular mechanism of Bushen Huoxue Formula from Shangke Dacheng （The Great Compendium of External Medicine） on tendon-bone healing after anterior cruciate ligament （ACL） injury. Methods The TCMSP database was used to screen active components and corresponding targets of the Chinese herbs in Bushen Huoxue Formula. Disease-related genes for tendon-bone healing after ACL injury were retrieved from GeneCards，OMIM，and PharmGkb databases. Common targets between active components and the disease were identified. Cytoscape software was used to construct a drug-active component-disease target network， and the String online platform was employed to build a protein-protein interaction （PPI） network. CytoNCA plugin in Cytoscape was used for network topology analysis to screen core targets and corresponding active components. Gene Ontology （GO） biological function and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analyses were performed on the common targets. Molecular docking between the top 5 active components （by degree value） and core targets was conducted using Autodocktools software，and the results were visualized. Results （1） The TCMSP database screening identified 121 effective active components and 243 potential targets from the Chinese herbs in Bushen Huoxue Formula. A total of 416 disease targets for tendon-bone healing after ACL injury were obtained from GeneCards （413），OMIM （3），and PharmGkb （4） databases. Forty-two common targets between drug active components and the disease were identified， corresponding to 108 active components. Network topology analysis screened 16 core targets in the PPI network，including VEGFA，HIF1A， IL1B， IL6， IL10， TNF， EGF， TP53， CXCL8， CD44， SPP1， PTGS2， MMP2， MMP3， MMP9， and VCAM1. GO biological functions were enriched in extracellular matrix collagen metabolism， reactive oxygen species regulation，endopeptidase and cytokine activity，and integrin binding. KEGG pathways were enriched in vascular-related signaling pathways such as atherosclerosis and HIF-1，inflammation-related pathways including TNF，Toll-like receptor，and NF-κB，and immune-related pathways such as C-type lectin receptor. Based on KEGG enrichment results， core targets closely related to angiogenesis （VEGFA， HIF1A） and inflammation （IL1B， IL6， IL10， TNF， PTGS2） were selected for molecular docking. The results showed that key active components of Bushen Huoxue Formula had good binding activity with key targets for tendon-bone healing after ACL injury. Conclusion Bushen Huoxue Formula may promote tendon-bone healing after ACL injury by regulating angiogenesis and inflammation through active components such as quercetin，luteolin，kaempferol，β-carotene， and ellagic acid acting on targets including VEGFA，HIF1A，IL1B，IL6，IL10，TNF，and PTGS2.

R285.5

国家自然科学基金面上项目（编号：82274557）；广东省中医药局科研项目（编号：20231295）；深圳市“医疗卫生三名工程”项目 （编号：SZZYSM202101005）