Turcz., a traditional Chinese herbal medicine, displays good therapeutic efficiency against respiratory diseases (e.g. pneumonia, pharyngitis) in clinical applications, however, its effects on COPD and the mechanism of action are still unclear. This study aims to investigate the therapeutic effect of the ethyl acetate fraction of Turcz. (TCEA) on COPD and reveal the underlying mechanism. A cigarette smoke (CS)-induced mouse COPD model was established, and the efficacy of TCEA was evaluated using peripheral blood testing, HE and Masson staining, qRT-PCR and ELISA assays. TCEA was analyzed for chemical composition by LC-MS/MS and HPLC. Prediction of major signaling pathways and potential targets was performed by network pharmacology. The molecular mechanism of TCEA was explored by immunoblotting, immunofluorescence staining, flow cytometry, and ubiquitination assay. Finally, potential active small molecules in TCEA were identified by molecular virtual screening. TCEA treatment significantly inhibited the secretion of pro-inflammatory factors and attenuated pathological emphysema. The main chemical constituents of TCEA were identified as flavonoids by UPLC-MS/MS. Network pharmacology analysis enriched the Nrf2 signaling pathway closely related to oxidative stress. Our results suggested that TCEA inhibited ferroptosis by activating Nrf2/SLC7A11/GPX4 axis and inhibiting lipid metabolism-related proteins, ACSL4, ALOX5 and COX2 . Noteworthily, the beneficial impact of TCEA on regulation of SLC7A11 and GPX4 vanished after silencing Nrf2. Moreover, Nrf2 ubiquitination was inhibited by TCEA treatment. Finally, several flavonoids modulating Nrf2 were identified by molecular virtual screening. TCEA significantly alleviated COPD progression by inhibiting ferroptosis primarily through activation of Nrf2/SLC7A11/GPX4 signaling. Flavonoids are the main active components that exert their effects. These findings shed light on the mechanism of action of TCEA and its potential active components, providing a feasible approach for the treatment of COPD.

中文翻译：

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Thesium chinense Turcz 的乙酸乙酯部分。通过激活 Nrf2/SLC7A11/GPX4 轴介导的铁死亡的抑制来缓解慢性阻塞性肺疾病


中草药Turcz.在临床应用中对肺炎、咽炎等呼吸系统疾病显示出良好的治疗效果，但其对COPD的作用及作用机制尚不清楚。本研究旨在探讨 Turcz 的乙酸乙酯部分的治疗效果。 （TCEA）研究 COPD 并揭示其潜在机制。建立香烟烟雾（CS）诱导的小鼠慢性阻塞性肺病模型，并通过外周血检测、HE和Masson染色、qRT-PCR和ELISA测定评估TCEA的疗效。通过 LC-MS/MS 和 HPLC 分析 TCEA 的化学成分。通过网络药理学对主要信号通路和潜在靶标进行预测。通过免疫印迹、免疫荧光染色、流式细胞术和泛素化实验探讨了TCEA的分子机制。最后，通过分子虚拟筛选鉴定了TCEA中潜在的活性小分子。 TCEA治疗显着抑制促炎因子的分泌并减轻病理性肺气肿。通过UPLC-MS/MS鉴定TCEA的主要化学成分为黄酮类化合物。网络药理学分析丰富了与氧化应激密切相关的Nrf2信号通路。我们的结果表明，TCEA 通过激活 Nrf2/SLC7A11/GPX4 轴并抑制脂质代谢相关蛋白 ACSL4、ALOX5 和 COX2 来抑制铁死亡。值得注意的是，在沉默 Nrf2 后，TCEA 对 SLC7A11 和 GPX4 调节的有益影响消失了。此外，TCEA 处理抑制了 Nrf2 泛素化。最后，通过分子虚拟筛选鉴定了几种调节Nrf2的黄酮类化合物。 TCEA 主要通过激活 Nrf2/SLC7A11/GPX4 信号传导来抑制铁死亡，从而显着缓解 COPD 进展。黄酮类化合物是发挥其作用的主要活性成分。这些发现揭示了TCEA及其潜在活性成分的作用机制，为COPD的治疗提供了可行的方法。