基于网络药理学和分子对接探讨护骨胶囊治疗骨质疏松症的作用机制

目的 基于网络药理学和分子对接技术探讨护骨胶囊治疗骨质疏松症潜在的活性成分和作用机制。方法 利用TCMSP数据库结合《中国药典》2020版筛选候选活性成分，并通过文献检索进行补充和删减得到最终成分，并通过Swiss Target Prediction数据库预测和筛选出活性成分的潜在的作用靶点。通过权威数据库（Genecard、Drugbank、TTD）检索骨质疏松症相关靶点，通过Cytoscape 3.9.0软件建立中药–成分–靶点网络模型进行网络分析，并将交集靶点导入至String在线数据库进行蛋白相互作用（PPI）网络构建并进行分析。利用David数据库进行基因本体论（GO）和京都基因与基因组百科全书（KEGG）通路富集分析。利用AutoDockTools 1.5.6软件进行分子对接。结果 共筛选出护骨胶囊59个活性成分、78个交集靶点。PPI网络分析表明，肿瘤坏死因子（TNF）、血管内皮生长因子A（VEGFA）、非受体酪氨酸激酶（SRC）、酪氨酸蛋白激酶3（MAPK3）、雌激素受体α（ESR1）等可能是护骨胶囊治疗骨质疏松症的关键靶点。GO和KEGG富集分析表明护骨胶囊主要通过细胞外空间、质膜、宿主细胞核等细胞组分参与细胞生存、生长、分裂等众多生物学过程，继而与蛋白质特异性结合，发挥调节核受体活性、过渡金属离子结合、类固醇结合等分子功能，涉及糖尿病并发症中的晚期糖基化终末化产物（AGE）-晚期糖基化终末产物受体（RAGE）信号通路、松弛素信号通路、雌激素信号通路等。分子对接结果表明，地黄苦苷元、大黄素甲醚、甲基异茜草素、阿魏酸和大豆苷元核心成分与TNF、VEGFA、SRC、MAPK3、ESR1核心靶点均具有良好的结合能力。结论 护骨胶囊中的地黄苦苷元、大黄素甲醚、甲基异茜草素、阿魏酸和大豆苷元等可能是治疗骨质疏松症的物质基础，能够作用于TNF、VEGFA、SRC、MAPK3、ESR1等多个靶点，通过调节糖尿病并发症中的AGE-RAGE信号通路、松弛素信号通路、雌激素信号通路等参与骨骼重塑、抑制破骨细胞的形成与分化、促进骨形成和抗炎作用，进而发挥治疗骨质疏松症的作用。

Mechanism of action of Hugu Capsules in treatment of osteoporosis based on network pharmacology and molecular docking

Objective To explore the potential active ingredients and mechanism of Hugu Capsules in treating osteoporosis based on network pharmacology and molecular docking technology. Methods The candidate active ingredients were screened using TCMSP database in combination with the 2020 edition of Chinese Pharmacopoeia, and the final ingredients were obtained through supplement and deletion through literature search, and the potential targets of active ingredients were predicted and screened through the Swiss Target Prediction database. The targets related to osteoporosis were searched through authoritative databases (Genecard, Drugbank, TTD), and the TCM - component - target network model was established by Cytoscape 3.9.0 software for network analysis. The intersection targets were imported into the String online database for protein interaction (PPI) network construction and analysis. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed using the David database. Molecular docking was performed using AutoDockTools 1.5.6 software. Results A total of 59 active components and 78 intersection targets of Hugu Capsules were screened out. PPI network analysis showed that TNF, VEGFA, SRC, MAPK3, and ESR1 may be the key targets of Hugu Capsules in treatment of osteoporosis. GO and KEGG enrichment analysis showed that Hugu Capsules mainly participated in many biological processes such as cell survival, growth and division through extracellular space, plasma membrane, host nucleus and other cell components, and then specifically bound to proteins to regulate nuclear receptor activity, transition metal ion binding, steroid binding and other molecular functions, involving AGE-RAGE signaling pathway, relaxin signaling pathway and estrogen signaling pathway in diabetic complications. The results of molecular docking showed that the core components such as rehmannia aglycone, physcion, methyl isoalizarin, ferulic acid, and daidzein had good binding ability to core targets such as TNF, VEGFA, SRC, MAPK3, and ESR1. Conclusion Rehmanniogenin, physcion, methylisoalizarin, ferulic acid, and daidzein in Hugu Capsules may be the material basis for the treatment of osteoporosis, which can act on multiple targets such as TNF, VEGFA, SRC, MAPK3, and ESR1, and participate in bone remodeling, inhibit the formation and differentiation of osteoclasts, promote bone formation and anti-inflammatory effects by regulating AGE-RAGE signaling pathway, relaxin signaling pathway and estrogen signaling pathway in diabetic complications, and then play a role in treatment of osteoporosis.