Department of Pulmonary and Critical Care Medicine, National Regional Center for Respiratory Medicine, Jiangxi Hospital of China-Japan Friendship Hospital, The 1st Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, Jiangxi, China.
Department of Emergency, China-Japan Friendship Hospital, Beijing, China.
Phytother Res. 2024 Jul;38(7):3401-3416. doi: 10.1002/ptr.8212. Epub 2024 Apr 26.
Our previous research confirmed that rutin reduced ventilator-induced lung injury (VILI) in mice. Ferroptosis has been reported to participate in the pathogenic process of VILI. We will explore whether rutin inhibits ferroptosis to alleviate VILI. A mouse model of VILI was constructed with or without rutin pretreatment to perform a multiomics analysis. Hematoxylin-eosin (HE) staining and transmission electron microscopy were used to evaluate lung injury in VILI mice. Dihydroethidium (DHE) staining and the malondialdehyde (MDA) and superoxide dismutase (SOD) levels were detected. Molecular docking was performed to determine the binding affinity between rutin and ferroptosis-related proteins. Western blot analysis, real-time PCR (RT-PCR) and immunohistochemical (IHC) staining were conducted to detect the expression levels of GPX4, XCT, ACSL4, FTH1, AKT and p-AKT in lung tissues. Microscale thermophoresis (MST) was used to evaluate the binding between rutin and AKT1. Transcriptomic and proteomic analyses showed that ferroptosis may play a key role in VILI mice. Metabolomic analysis demonstrated that rutin may affect ferroptosis via the AKT pathway. Molecular docking analysis indicated that rutin may regulate the expression of ferroptosis-related proteins. Moreover, rutin upregulated GPX4 expression and downregulated the expression of XCT, ACSL4 and FTH1 in the lung tissues. Rutin also increased the ratio of p-AKT/AKT and p-AKT expression. MST analysis showed that rutin binds to AKT1. Rutin binds to AKT to activate the AKT signaling pathway, contributing to inhibit ferroptosis, thus preventing VILI in mice. Our study elucidated a possible novel strategy of involving the use of rutin for preventing VILI.
我们之前的研究证实，芦丁可减轻小鼠呼吸机诱导的肺损伤（VILI）。铁死亡已被报道参与 VILI 的发病过程。我们将探讨芦丁是否通过抑制铁死亡来减轻 VILI。使用或不使用芦丁预处理构建 VILI 小鼠模型进行多组学分析。苏木精-伊红（HE）染色和透射电子显微镜用于评估 VILI 小鼠的肺损伤。二氢乙啶（DHE）染色和丙二醛（MDA）和超氧化物歧化酶（SOD）水平检测。分子对接用于确定芦丁与铁死亡相关蛋白的结合亲和力。Western blot 分析、实时 PCR（RT-PCR）和免疫组织化学（IHC）染色用于检测肺组织中 GPX4、XCT、ACSL4、FTH1、AKT 和 p-AKT 的表达水平。微量热泳动（MST）用于评估芦丁与 AKT1 的结合。转录组和蛋白质组学分析表明，铁死亡可能在 VILI 小鼠中发挥关键作用。代谢组学分析表明，芦丁可能通过 AKT 通路影响铁死亡。分子对接分析表明，芦丁可能调节铁死亡相关蛋白的表达。此外，芦丁上调肺组织中 GPX4 的表达，下调 XCT、ACSL4 和 FTH1 的表达。芦丁还增加了 p-AKT/AKT 的比值和 p-AKT 的表达。MST 分析表明，芦丁与 AKT1 结合。芦丁与 AKT 结合激活 AKT 信号通路，有助于抑制铁死亡，从而防止小鼠发生 VILI。我们的研究阐明了一种新的潜在策略，即使用芦丁预防 VILI。
