College of First Clinical Medical, Shandong University of Traditional Chinese Medicine, Jinan, China.
Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan, China.
Chem Biol Drug Des. 2023 Dec;102(6):1489-1505. doi: 10.1111/cbdd.14340. Epub 2023 Sep 10.
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.
本研究采用网络药理学和分子对接技术预测黄芪（AR）促进骨髓间充质干细胞（BM-MSCs）成骨分化的活性成分和作用机制，并进行细胞实验验证。首先，采用网络药理学预测 AR 促进成骨分化的有效成分、作用靶点和作用机制。通过数据库收集 AR 的有效成分及相应的靶蛋白和成骨分化的靶蛋白，取两者的交集靶蛋白进行蛋白质-蛋白质相互作用（PPI）网络的构建和分析，并进行基因本体（GO）和京都基因与基因组百科全书（KEGG）富集分析。然后，采用分子对接技术验证活性成分与靶蛋白的相互作用，筛选合适的有效活性成分。最后，通过体外实验验证网络药理学分析结果。通过检索 AR 和成骨分化靶点的交集，共检索到 95 个潜在靶点。PPI 网络分析表明，RAC-α-丝氨酸-苏氨酸蛋白激酶（Akt1）被认为是 AR 调节成骨分化的最可靠靶标。GO 富集分析包括 21 个生物学过程、21 个细胞成分和 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发挥重要作用。分子对接结果表明，槲皮素的性能优于山奈酚。体外实验表明，槲皮素促进 BMSCs 中成骨标志物蛋白（包括胶原 I、Runt 相关转录因子 2 和骨桥蛋白）的表达，并激活 PI3K/Akt 信号通路。AR 通过其主要活性成分槲皮素作用于 Akt1 靶点，通过激活 PI3K/Akt 信号通路促进 BM-MSCs 的成骨分化。
