目的 通過網(wǎng)絡(luò)藥理學(xué)、分子對接和體外實驗探討淫羊藿素治療多發(fā)性骨髓瘤的作用機制。方法 利用PharmMapper、SEA、SwissTargetPrediction數(shù)據(jù)庫收集淫羊藿素的作用靶點，通過DisGeNET、GeneCards、MalaCards數(shù)據(jù)庫收集多發(fā)性骨髓瘤的疾病靶點。利用Venn分析獲得作用靶點與疾病靶點的共有靶點并導(dǎo)入STRING數(shù)據(jù)庫分析靶點間相互作用關(guān)系，再通過Cytoscape軟件構(gòu)建淫羊藿素與抗多發(fā)性骨髓瘤潛在作用靶點的可視化關(guān)系網(wǎng)絡(luò)，然后對潛在作用靶點進(jìn)行基因本體（GO）功能富集分析和和京都基因和基因組百科全書（KEGG）富集分析，并對關(guān)鍵靶點進(jìn)行分子對接驗證。進(jìn)一步對人多發(fā)性骨髓瘤U266細(xì)胞開展體外實驗，CCK-8法檢測淫羊藿素對U266細(xì)胞增殖的影響，qPCR和Western blotting檢測淫羊藿素對關(guān)鍵通路中相關(guān)基因、蛋白表達(dá)的影響。結(jié)果 篩選得到淫羊藿素治療多發(fā)性骨髓瘤的潛在作用靶點143個，其中的重要作用靶點主要有蛋白激酶B1（Akt1）、白蛋白（ALB）、連環(huán)蛋白β1（CTNNB1）、熱休克蛋白90α家族A類成員1（HSP90AA1）、表皮生長因子受體（EGFR）、促分裂原活化蛋白激酶1（MAPK1）、酪氨酸激酶受體2（ERBB2）、磷脂酰肌醇3-激酶調(diào)節(jié)亞基1（PIK3R1）。GO功能、KEGG通路富集分析和分子對接結(jié)果表明，淫羊藿素可能作用于PI3K/Akt信號通路。體外細(xì)胞實驗顯示，淫羊藿素可抑制人多發(fā)性骨髓瘤U266細(xì)胞增殖，并顯著上調(diào)U266細(xì)胞中PIK3R1表達(dá)、下調(diào)Akt1表達(dá)（P＜0.05、0.01）。結(jié)論 淫羊藿素可能通過調(diào)控PI3K/Akt信號通路中PIK3R1、Akt1的表達(dá)，抑制多發(fā)性骨髓瘤細(xì)胞增殖，發(fā)揮治療多發(fā)性骨髓瘤的潛在作用，為淫羊藿素的進(jìn)一步開發(fā)利用提供理論依據(jù)。

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.

R285

國家自然科學(xué)基金資助項目（82273954）