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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

32卷

期数:

2026年04期

页码:

362-370

栏目:

论著

出版日期:

2026-07-10

文章信息/Info

Title:

The occurrence of ferroptosis and its underlying molecular mechanisms in doxorubicin-induced myocardial injury

文章编号:

1006-8147（2026）04-0362-09

作者:

张怡梦; 张华璐; 徐哲龙

（天津医科大学基础医学院生理学与病理生理学系，天津 300070）

Author(s):

ZHANG Yimeng;  ZHANG Hualu; XU Zhelong

（Department of Pathophysiology， School of Basic Medical Sciences， Tianjin Medical University， Tianjin 300070， China）

关键词:

阿霉素; 铁死亡; 线粒体自噬; 谷胱甘肽过氧化物酶4

Keywords:

Doxorubicin;  ferroptosis; mitophagy; glutathione peroxidase 4

分类号:

R363

DOI:

10.20135/j.issn.1006-8147.2026.04.0362

文献标志码:

A

摘要:

目的：探讨阿霉素（doxorubicin，DOX）诱导心肌损伤过程中铁死亡的发生及分子机制。方法：采用腹腔注射DOX构建小鼠心肌损伤模型，分为对照（control组）和DOX处理组；用DOX处理HL-1细胞建立体外模型，分为对照（control组）和DOX处理组。通过RT-qPCR检测心肌组织和细胞的谷胱甘肽过氧化物酶（GPX）4、前列腺素内过氧化物合酶2（PTGS2） mRNA表达水平；采用GPX活性检测试剂盒检测GPX活性；通过乳酸脱氢酶（LDH）及丙二醛（MDA）检测试剂盒检测评估细胞损伤和脂质过氧化水平；采用苏木精-伊红（HE）染色法观察心脏病理损伤情况；利用MitoSOX Red、TMRE、BODIPY C11荧光探针分别检测线粒体活性氧簇（ROS）水平、线粒体膜电位及脂质过氧化物水平；利用CCK-8检测细胞活力；通过Western印迹法检测GPX4、LC3、P62、TOM20、TIM23、PINK1、Parkin等蛋白表达量。结果：RT-qPCR结果显示，与control组相比，DOX处理组HL-1细胞及小鼠心肌组织PTGS2 mRNA水平明显升高（t=8.281、5.065，均P＜0.01）。HE染色显示，DOX导致小鼠心肌纤维排列紊乱和结构破坏。LDH结果显示，与control组相比，DOX处理组显著加重心肌组织及HL-1细胞损伤（t=3.909、12.850，均P＜0.01）。共聚焦结果显示，与control组相比，DOX处理组HL-1细胞线粒体ROS升高（t=10.090，P＜0.01），线粒体膜电位降低（t=16.690，P＜0.01），脂质过氧化物水平升高（t=3.123，P＜0.05）。进一步检测发现，与control组相比，DOX处理组显著升高HL-1细胞及小鼠心肌组织中的MDA水平（t=3.431、4.509，P＜0.01），同时降低GPX活性（t=4.914，P＜0.01；t=2.505，P＜0.05）。Western印迹结果显示，与control组相比，DOX处理组HL-1细胞mito-GPX4、P62、mito-TOM20、mito-TIM23、mito-LC3、mito-P62蛋白表达明显下降（t=4.274、7.685、3.484、3.959、3.961、5.674，均P＜0.01），mito-PINK1、mito-Parkin、LC3蛋白表达水升高（t=4.111、5.317，均P＜0.01；t=2.631，P＜0.05）；与control组相比，DOX处理组小鼠心肌组织mito-GPX4、P62、mito-TIM23、mito-TOM20蛋白表达明显下降（t=5.486、11.58、5.946，均P＜0.01；t=3.640，P＜0.05），LC3蛋白表达水升高（t=3.567，P＜0.01）。结论：DOX通过过度激活线粒体自噬，导致GPX4蛋白水平降低，抗氧化能力受损，促进脂质过氧化物积累，进而诱导心肌细胞铁死亡并加重心肌损伤。

Abstract:

Objective： To investigate the occurrence of ferroptosis and its underlying molecular mechanisms in doxorubicin（DOX）-induced myocardial injury. Methods：A mouse model of myocardial injury was established by intraperitoneal injection of DOX， and animals were divided into control and DOX-treated groups. An in vitro model was generated by treating HL-1 cardiomyocytes with DOX， and cells were divided into control and DOX-treated groups. RT-qPCR was performed to measure the mRNA expression levels of glutathione peroxidase （GPX）4 and PTGS2 in myocardial tissue and cells. GPX activity was determined using a GPX activity assay kit. Myocardial and cellular injury was evaluated by lactate dehydrogenase （LDH） assay kits， while lipid peroxidation was assessed by malondialdehyde （MDA） assay kits. Histopathological changes in cardiac tissue were examined by hematoxylin and eosin （HE） sta-ining. MitoSOX Red， TMRE， and BODIPY C11 fluorescent probes were used to detect mitochondrial reactive oxygen species （ROS）， mitochondrial membrane potential， and lipid peroxidation levels， respectively. Cell viability was assessed using the CCK-8 assay. Protein expression levels of GPX4， LC3， P62， TOM20， TIM23， PINK1， and Parkin were analyzed by Western blotting. Results： RT-qPCR analysis demonstrated that compared with control group， PTGS2 mRNA expression was significantly increased in DOX-treated HL-1 cells and mouse myocardial tissue （t=8.281， 5.065， both P<0.01）. HE staining revealed disorganized myocardial fiber arrangement and marked structural damage following DOX administration. LDH assays showed that compared with control group， DOX markedly exacerbated myocardial and cellular injury（t=3.909， 12.850， both P<0.01）. Confocal microscopy revealed that compared with control group， DOX treatment significantly increased mitochondrial ROS production， reduced mitochondrial membrane potential， and elevated lipid peroxidation levels in HL-1 cells （t=10.090， 16.690， all P<0.01； t=3.123， P<0.05）. Consistently， compared with control group， MDA levels were significantly increased， whereas GPX activity was markedly decreased in both HL-1 cells and mouse myocardial tissue after DOX exposure （t=3.431， 4.509， 4.914， all P<0.01；t=2.505， P<0.05）. Western blotting further demonstrated that compared with control group， DOX treatment significantly reduced the protein expression of mito-GPX4， P62， mito-TOM20， mito-TIM23， mito-LC3， and mito-P62 in HL-1 cells（t=4.274， 7.685， 3.484，3.959，3.961，5.674， all P＜0.01）， while increased the expression of mito-PINK1， mito-Parkin， and LC3 protein（t=4.111， 5.317， both P＜0.01； t=2.631， P＜0.05）. In mouse myocardial tissue，compared with control group， DOX treatment significantly reduced the protein expression of mito-GPX4， P62， mito-TIM23， and mito-TOM20（t=5.486， 11.58，5.946， all P＜0.01； t=3.640， P＜0.05）， while increased LC3 protein expression（t=3.567， P＜0.01）. Conclusion： DOX induces ferroptosis in cardiomyocytes by excessively activating mitophagy， leading to mito-GPX4 depletion， impaired antioxidant capacity， and excessive lipid peroxidation， thereby exacerbating myocardial injury.

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备注/Memo

备注/Memo:

（2026-01-05收稿）
基金项目 天津市教委科研计划项目（2022KJ195）
作者简介 张怡梦（2000-），女，硕士在读，研究方向：病理学与病理生理学；通信作者：徐哲龙，E-mail：zxu@tmu.edu.cn。

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更新日期/Last Update: 2026-07-15