目的 通过网络药理学、分子对接技术和斑马鱼实验探究仙鹤草治疗糖皮质激素性骨质疏松症可能的作用机制。方法 通过中药系统药理学数据库与分析平台（TCMSP）和本草组鉴（HERB）数据库初步筛查出仙鹤草相关的活性成分；利用SEA和SwissTargetPrediction平台预测活性成分的相关靶点；利用OMIM、GeneCards数据库检索糖皮质激素性骨质疏松症相关的疾病靶点；利用韦恩图绘制平台获取共有的关键作用靶点，并将信息导入Cytoscope 3.7.2软件和STRING在线分析平台，进行可视化分析；使用DAVID数据库进行基因本体（GO）和京都基因与基因组百科全书（KEGG）富集分析；然后通过Autodock Tools 1.5.6软件对有效成分-核心作用靶点进行分子对接验证。通过斑马鱼幼鱼骨形成抑制模型探讨仙鹤草水提液对糖皮质激素所致的斑马鱼幼鱼骨形成抑制的影响及机制。结果 通过TCMSP和HERB数据库筛选出仙鹤草的32种潜在活性成分，通过SEA和SwissTargetPrediction平台预测并筛选后得277个药物靶点；通过OMIM和GeneCards数据库检索糖皮质激素性骨质疏松症相关的潜在疾病靶点共654个。将药物-疾病的36个交集靶点进行富集分析，GO功能富集主要包括通过RNA聚合酶Ⅱ的转录正调控、信号转导、DNA模板的正向调控等；KEGG富集通路主要富集在磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（Akt）信号通路等信号通路。将degree值前5个的有效成分（槲皮素、木犀草素、鞣花酸、芹菜素和山柰酚）与核心靶点[丝氨酸/苏氨酸蛋白激酶1（Akt1）、非受体酪氨酸激酶（SRC）、雌激素受体1（ESR1）、基质金属蛋白酶-9（MMP9）、核因子κB亚基1（NFKB1）、糖原合成酶激酶-3β（GSK-3β）、前列腺素内过氧化物合酶2（PTGS2）和Jun原癌基因]进行分子对接，结果显示仙鹤草的5种活性成分与糖皮质激素性骨质疏松症核心靶点的结合活性均较好。通过斑马鱼幼鱼骨形成抑制模型验证仙鹤草水提液可部分通过PI3K/Akt信号通路缓解糖皮质激素所致的斑马鱼幼鱼骨形成抑制的现象。结论 揭示了仙鹤草治疗糖皮质激素性骨质疏松症的有效成分和作用机制，为仙鹤草治疗糖皮质激素性骨质疏松症的临床实践及后续研究提供参考。

Objective To explore the mechanism of action of Agrimoniae Herba in treatment of glucocorticoid-induced osteoporosis based on network pharmacology, molecular docking technology, and zebrafish experiments. Methods Preliminary screening of active ingredients related to Agrimoniae Herba was conducted through the TCMSP and the HERB database; Using SEA and SwissTarget Prediction platforms to predict the relevant targets of active ingredients. Retrieve disease targets related to glucocorticoid induced osteoporosis using OMIM and GeneCards databases. Using the Venn diagram drawing platform to obtain common key targets, and importing the information into Cytoscope 3.7.2 software and STRING online analysis platform for visualization analysis. Perform GO and KEGG enrichment analysis using the DAVID database. Molecular docking validation was performed on the active ingredient core target using Autodock Tools 1.5.6 software. Through a zebrafish larval bone formation inhibition model, the effects and mechanisms of the water extract of Agrimoniae Herba on glucocorticoid-induced inhibition of bone formation in zebrafish larvae were explored. Results 32 Active ingredients of Agrimoniae Herba were screened through TCMSP and HERB databases, with the top 5 core active ingredients (quercetin, luteolin, ellagic acid, apigenin, and kaempferol). After prediction and screening using SEA and SwissTarget Prediction platforms, 277 drug targets were identified. A total of 654 potential disease targets related to glucocorticoid induced osteoporosis were retrieved through OMIM and GeneCards databases. Enrichment analysis of the 36 intersecting targets of drugs and diseases showed that GO functional enrichment mainly included positive regulation of transcription by RNA polymerase II, signal transduction, and positive regulation of DNA-templated processes. KEGG enrichment pathways were mainly focused on the PI3K/Akt signaling pathway. Molecular docking of the top 5 effective components (quercetin, luteolin, ellagic acid, apigenin, and kaempferol) with the core targets (Akt1, SRC, ESR1, MMP9, NFKB1, GSK-3β, PTGS2, and JUN) showed had good binding activity. The zebrafish larval bone formation inhibition model verified that the water extract of Agrimoniae Herba could partially alleviate the glucocorticoid-induced inhibition of bone formation in zebrafish larvae through the PI3K/Akt signaling pathway. Conclusion This study preliminarily revealed the effective ingredients and mechanism of action of Agrimoniae Herba in treatment of glucocorticoid-induced osteoporosis, providing reference for the clinical practice and subsequent research of Agrimoniae Herba in treatment of glucocorticoid-induced osteoporosis.

R285

广东省医学科学技术研究基金资助项目（A2024138）；广东省药学会科学研究基金资助项目（2023QNTJ39）