目的 通过网络药理学、分子对接和体外实验探讨淫羊藿素治疗多发性骨髓瘤的作用机制。方法 利用PharmMapper、SEA、SwissTargetPrediction数据库收集淫羊藿素的作用靶点，通过DisGeNET、GeneCards、MalaCards数据库收集多发性骨髓瘤的疾病靶点。利用Venn分析获得作用靶点与疾病靶点的共有靶点并导入STRING数据库分析靶点间相互作用关系，再通过Cytoscape软件构建淫羊藿素与抗多发性骨髓瘤潜在作用靶点的可视化关系网络，然后对潜在作用靶点进行基因本体（GO）功能富集分析和和京都基因和基因组百科全书（KEGG）富集分析，并对关键靶点进行分子对接验证。进一步对人多发性骨髓瘤U266细胞开展体外实验，CCK-8法检测淫羊藿素对U266细胞增殖的影响，qPCR和Western blotting检测淫羊藿素对关键通路中相关基因、蛋白表达的影响。结果 筛选得到淫羊藿素治疗多发性骨髓瘤的潜在作用靶点143个，其中的重要作用靶点主要有蛋白激酶B1（Akt1）、白蛋白（ALB）、连环蛋白β1（CTNNB1）、热休克蛋白90α家族A类成员1（HSP90AA1）、表皮生长因子受体（EGFR）、促分裂原活化蛋白激酶1（MAPK1）、酪氨酸激酶受体2（ERBB2）、磷脂酰肌醇3-激酶调节亚基1（PIK3R1）。GO功能、KEGG通路富集分析和分子对接结果表明，淫羊藿素可能作用于PI3K/Akt信号通路。体外细胞实验显示，淫羊藿素可抑制人多发性骨髓瘤U266细胞增殖，并显著上调U266细胞中PIK3R1表达、下调Akt1表达（P＜0.05、0.01）。结论 淫羊藿素可能通过调控PI3K/Akt信号通路中PIK3R1、Akt1的表达，抑制多发性骨髓瘤细胞增殖，发挥治疗多发性骨髓瘤的潜在作用，为淫羊藿素的进一步开发利用提供理论依据。

Objective To investigate the mechanism of icaritin in treatment of multiple myeloma through network pharmacology, molecular docking and in vitro experiments. Methods To collect the targets of icaritin using PharmaMapper, SEA, and SwissTargetPrediction databases, and collect disease targets of multiple myeloma using DisGeNET, GeneCards, and MalaCards databases. Using Venn analysis to obtain common targets for action and disease targets, and importing them into the STRING database to analyze the interaction relationships between targets. Then, using Cytoscape software, a visual relationship network between icaritin and potential targets for anti multiple myeloma action is constructed. Then, GO and KEGG enrichment analysis are performed on potential targets, and molecular docking verification is performed on key targets. Further in vitro experiments were conducted on human multiple myeloma U266 cells. The CCK-8 method was used to detect the effect of icaritin on U266 cell proliferation, and qPCR and Western blotting were used to detect the effect of icaritin on the expression of related genes and proteins in key pathways. Results A total of 143 potential targets of icaritin in treatment of multiple myeloma were selected. The important targets were Akt1, ALB, CTNNB1, HSP90AA1, EGFR, MAPK1, ERBB2, and PIK3R1. GO and KEGG enrichment and molecular docking results showed that icaritin may affect the PI3K/Akt signaling pathway. In vitro experiments showed that icaritin could inhibit the proliferation of human multiple myeloma U266 cells, and significantly up-regulate the expression of PIK3R1 and down-regulate the expression of Akt1 in U266 cells (P < 0.05, 0.01). Conclusion Icaritin could inhibit the proliferation of multiple myeloma cells by regulating the expression of PIK3R1 and Akt1 in the PI3K/Akt signaling pathway, and play a potential role in the treatment of multiple myeloma, providing a theoretical basis for the further development and utilization of icaritin.

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国家自然科学基金资助项目（82273954）