黄芩苷镁诱导三阴性乳腺癌细胞铁死亡的作用研究

Study on the role of baicalin magnesium in inducing ferroptosis in triple-negative breast cancer cells

摘要

目的 结合网络药理学与分子对接技术筛选黄芩苷镁诱导三阴性乳腺癌（TNBC）细胞铁死亡的潜在作用靶点,并探讨其可能的作用机制。方法 通过CTD、HERB、GeneCards、OMIM及FerrDb数据库筛选黄芩苷镁、TNBC及铁死亡相关靶点,采用Venny获取三者交集基因,结合STRING、Cytoscape进行PPI网络分析,并开展GO/KEGG富集及分子对接分析,筛选黄芩苷镁诱导TNBC细胞铁死亡的潜在关键靶点。以不同浓度黄芩苷镁（50、100、200 μg·mL^-1）处理TNBC细胞MDA-MB-231,检测细胞活力,以及细胞内还原型谷胱甘肽（GSH）、Fe²⁺、脂质过氧化物和活性氧（ROS）含量/水平,并检测铁死亡相关蛋白表达。结果 网络药理学分析获得57个黄芩苷镁-TNBC共同作用靶点,与铁死亡相关基因进一步交集获得21个候选靶点。PPI网络及拓扑分析筛选出关键靶点,包括Kelch样ECH关联蛋白1（KEAP1,Keap1）、核因子E2相关因子2（NFE2L2,Nrf2）以及血红素加氧酶-1（HMOX1,HO-1）等铁死亡调控相关基因。分子对接结果显示,黄芩苷镁与Keap1、Nrf2及HO-1具有一定结合能力。体外实验发现,3个浓度的黄芩苷镁均可降低MDA-MB-231细胞活力和GSH水平,升高MDA-MB-231细胞Fe²⁺、脂质过氧化物和ROS含量/水平;下调MDA-MB-231细胞Nrf2、HO-1、GPX4蛋白表达,上调Keap1蛋白表达。结论 通过网络药理学及分子对接分析筛选黄芩苷镁诱导TNBC细胞铁死亡的潜在关键靶点及作用通路,黄芩苷镁可能通过调控Keap1/Nrf2/HO-1轴促进MDA-MB-231细胞铁死亡。

Abstract

Objective To screen the potential targets of baicalin magnesium in inducing ferroptosis in triple-negative breast cancer (TNBC) cells using network pharmacology and molecular docking, and to investigate its potential mechanisms. Methods CTD, HERB, GeneCards, OMIM, and FerrDb databases were used to screen the targets related to baicalin magnesium, TNBC, and ferroptosis; Venny was used to obtain the intersection genes among the three. PPI network analysis was conducted using STRING and Cytoscape, as well as GO and KEGG enrichment analyses and molecular docking to screen potential key targets of baicalin magnesium-induced ferroptosis in TNBC cells. TNBC cells MDA-MB-231 were treated with baicalin magnesium at different concentrations (50, 100, 200 μg·mL^-1). The cell viability, as well as intracellular glutathione (GSH), Fe²⁺, lipid peroxides, reactive oxygen species (ROS) content/level, and ferroptosis-related proteins were detected. Results Network pharmacological analysis identified 57 common targets associated with baicalin magnesium and TNBC, of which 21 candidate targets were obtained by further intersection with ferroptosis-related genes. PPI network and topological analyses screened key targets, including Kelch-like ECH-associated protein 1 (KEAP1), nuclear factor erythroid 2-related factor 2 (NFE2L2, Nrf2), and heme oxygenase-1 (HMOX1, HO-1), which are related to ferroptosis regulation. Molecular docking analysis showed that baicalin magnesium exhibited a certain binding ability toward Keap1, Nrf2, and HO-1. In vitro experiments showed that baicalinmagnesium at all three concentrations reduced cell viability and GSH content in MDA-MB-231 cells, while increasing the levels of Fe²⁺, lipid peroxides and ROS. It down-regulated the protein expression of Nrf2, HO-1 and GPX4, and upregulated Keap1 protein expression. Conclusion Network pharmacological and molecular docking analyses identified potential key targets and signaling pathways involved in baicalin magnesium-induced ferroptosis in TNBC cells, baicalin magnesium may promote ferroptosis in MDA-MB-231 cells by regulating the Keap1/Nrf2/HO-1 axis.

关键词

黄芩苷镁 / 网络药理学 / 三阴性乳腺癌 / 铁死亡 / Keap1/Nrf2/HO-1轴 / 氧化应激

Key words

baicalin magnesium / network pharmacology / triple-negative breast cancer / ferroptosis / Keap1/Nrf2/HO-1 axis / oxidative stress

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基金资助

承德医学院校级课题（KY202601）