Zheng Hao, Ou Jinbo, Han Hui, Lu Qizheng, Shen Yunli
Department of Critical Care Medicine, Zhongda Hospital, School of Medicine, Southeast University, No.87, Dingjiaqiao, Gulou District, Nanjing 210009, China; Department of Cardiology, Shanghai East Hospital, Tongji University School of Medicine, Shanghai 200120, China.
Departments of Cardiology, Fudan University Zhongshan Hospital, Qingpu Branch, 1158 Park East Road, Shanghai 60518120, China.
Biomed Pharmacother. 2025 Feb;183:117832. doi: 10.1016/j.biopha.2025.117832. Epub 2025 Jan 22.
Targeting mitochondrial ferroptosis presents a promising strategy for mitigating myocardial ischemia-reperfusion (I/R) injury. This study aims to evaluate the efficacy of the mitochondrial-targeted ferroptosis inhibitor SS-31@Fer-1 (elamipretide@ferrostatin1) in reducing myocardial I/R injury.
SS-31@Fer-1 was synthesized and applied to H9C2 cells subjected to hypoxia/reoxygenation (H/R) to assess its protective effects. Cytotoxicity was evaluated using a cell counting kit-8 (CCK-8) assay, with lactate dehydrogenase (LDH) and creatine kinase isoenzyme (CK-MB) levels measured. Mitochondrial reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) were assessed using Mito-SOX and JC-1 fluorescent dyes, respectively. Lipid peroxidation products, malondialdehyde (MDA) and glutathione (GSH), were quantified. Mitochondrial structure, mt-cytochrome b (mt-Cytb), and mt-ATP synthase membrane subunit 6 (mt-ATP6) were analyzed. Additionally, iron homeostasis and ferroptosis markers were evaluated.
SS-31@Fer-1 significantly improved H/R-induced cardiomyocyte viability and reduced LDH and CK-MB levels. Compared to the Fer-1 group, SS-31@Fer-1 reduced GSH and increased MDA levels, enhancing mitochondrial integrity and function. Notably, it increased mitochondrial ROS and decreased MMP, indicating a mitigation of H/R-induced cardiomyocyte cytotoxicity. Furthermore, SS-31@Fer-1 maintained cellular iron homeostasis, as evidenced by increased expression of FTH, FTMT, FPN, and ABCB8. Elevated levels of GPX4 and Nrf2 were observed, while ACSL4 and PTGS2 levels were reduced in the SS-31@Fer-1 group.
SS-31@Fer-1 effectively suppressed ferroptosis in H/R-induced cardiomyocytes by maintaining cellular iron homeostasis, improving mitochondrial function, and inhibiting oxidative stress. These findings provide novel insights and opportunities for alleviating myocardial I/R injury.
靶向线粒体铁死亡是减轻心肌缺血再灌注（I/R）损伤的一种有前景的策略。本研究旨在评估线粒体靶向铁死亡抑制剂SS-31@Fer-1（依拉米肽@铁抑素1）在减轻心肌I/R损伤方面的疗效。
合成SS-31@Fer-1并将其应用于经历缺氧/复氧（H/R）的H9C2细胞，以评估其保护作用。使用细胞计数试剂盒-8（CCK-8）测定法评估细胞毒性，测量乳酸脱氢酶（LDH）和肌酸激酶同工酶（CK-MB）水平。分别使用Mito-SOX和JC-1荧光染料评估线粒体活性氧（ROS）和线粒体膜电位（MMP）。对脂质过氧化产物丙二醛（MDA）和谷胱甘肽（GSH）进行定量。分析线粒体结构、线粒体细胞色素b（mt-Cytb）和线粒体ATP合酶膜亚基6（mt-ATP6）。此外，评估铁稳态和铁死亡标志物。
SS-31@Fer-1显著提高了H/R诱导的心肌细胞活力，并降低了LDH和CK-MB水平。与Fer-1组相比，SS-31@Fer-1降低了GSH水平，增加了MDA水平，增强了线粒体完整性和功能。值得注意的是，它增加了线粒体ROS并降低了MMP，表明减轻了H/R诱导的心肌细胞毒性。此外，SS-31@Fer-1维持了细胞铁稳态，FTH、FTMT、FPN和ABCB8表达增加证明了这一点。在SS-31@Fer-1组中观察到GPX4和Nrf2水平升高，而ACSL4和PTGS2水平降低。
SS-31@Fer-1通过维持细胞铁稳态、改善线粒体功能和抑制氧化应激，有效抑制了H/R诱导的心肌细胞中的铁死亡。这些发现为减轻心肌I/R损伤提供了新的见解和机会。
