本研究采用网络药理学和分子对接技术预测黄芪对骨髓间充质干细胞（BM-MSCs）成骨分化的活性成分和作用机制，并进行了细胞实验证实。首先，利用网络药理学预测黄芪促进成骨分化的有效成分、靶点和作用机制。通过数据库收集黄芪的有效成分及其相应的靶蛋白和成骨分化的靶蛋白，使用两者的共同靶标构建和分析蛋白质相互作用网络。进行基因本体论（GO）和Kyoto百科全书和基因组（KEGG）富集分析。接下来，利用分子对接技术验证活性成分与靶蛋白之间的相互作用，并选择合适的有效活性成分。最后，通过体外实验验证了网络药理学分析的结果。通过搜索黄芪和成骨分化靶标的交集，共检索到95个潜在靶标。蛋白质相互作用网络分析表明，RAC-α丝氨酸/苏氨酸蛋白激酶（Akt1）被认为是黄芪调节成骨分化的最可靠靶标。GO富集分析涵盖了21个生物过程、21个细胞组分和100个分子功能。KEGG富集分析表明，Ⅰ类磷脂酰肌醇-3激酶（PI3K）丝氨酸/苏氨酸激酶（Akt）信号通路可能在促进成骨分化中起到重要作用。分子对接的结果显示，槲皮素的效果比山奈酚更好。体外实验证明，槲皮素促进BMSCs中成骨标志蛋白（包括胶原I、Runt相关转录因子2和骨蛋白）的表达，并激活PI3K/Akt信号通路。黄芪通过其主要活性成分槲皮素作用于Akt1靶标，并通过激活PI3K/Akt信号通路促进BM-MSCs的成骨分化。© 2023 John Wiley & Sons Ltd.

The present study used network pharmacology and molecular docking to predict the active ingredients and mechanisms of action of Astragalus radix (AR) to promote osteogenic differentiation of bone marrow mesenchymal stem cells (BM-MSCs), and cell experiments were conducted for verification. First, network pharmacology was used to predict the effective components, targets, and mechanisms of action of AR to promote osteogenic differentiation. The effective components and corresponding target proteins of AR, and the target proteins of osteogenic differentiation were collected through the database. The intersection targets of the two were used for the construction and analysis of a protein-protein interaction (PPI) network. Gene Oncology (GO) and Kyoto Encyclopedia of Genes, and Genomes (KEGG) enrichment analyses were conducted. Next, molecular docking technology was carried out to verify the interaction between the active ingredient and the target protein, and to select the appropriate effective active ingredient. Finally, the results of network pharmacology analysis were verified by in vitro experiments. A total of 95 potential targets were retrieved by searching the intersection of AR and osteogenic differentiation targets. PPI network analysis indicated that RAC-α-serine-threonine-protein kinase (Akt1) was considered to be the most reliable target for AR to regulate osteogenic differentiation. GO enrichment analysis included 21 biological processes, 21 cellular components and 100 molecular functions. KEGG enrichment analysis indicated that the class I phosphatidylinositol-3 kinase (PI3K)-serine-threonine kinase (Akt) signaling pathway may play an important role in promoting osteogenic differentiation. The results of molecular docking showed that quercetin's performance was improved compared with that of kaempferol. In vitro experiments showed that quercetin promoted the expression of osteogenic marker proteins (including collagen I, Runt-related transcription factor 2 and osteopontin) in BMSCs and activated the PI3K/Akt signaling pathway. AR acted on Akt1 targets through its main active component quercetin, and promoted the osteogenic differentiation of BM-MSCs by activating the PI3K/Akt signaling pathway.© 2023 John Wiley & Sons Ltd.