Quantitative proteomic analyses reveal that GPX4 downregulation during myocardial infarction contributes to ferroptosis in cardiomyocytes
Quantitative proteomic analyses reveal that GPX4 downregulation during myocardial infarction contributes to ferroptosis in cardiomyocytes
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DOI：
10.1038/s41419-019-2061-8
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发表时间：
2019-11-04
影响因子：
9
通讯作者：
Lee, Sang Chul
中科院分区：
文献类型：
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作者：
Park, Tae-Jun;Park, Jei Hyoung;Lee, Sang Chul
缺血性心脏病（IHD）是世界范围内导致死亡的主要原因。虽然心肌细胞死亡在心肌梗死（MI）中起着重要的作用，但其潜在的机制仍有待阐明。为了了解心肌梗死的进展并确定潜在的治疗靶点，我们使用心肌梗死小鼠模型进行了基于串联质量标签（TMT）的定量蛋白质组学分析。基因本体学（GO）分析和基因集富集分析（GSEA）显示，心肌梗死期间谷胱甘肽代谢途径和活性氧（ROS）途径显著下调，尤其是保护细胞不发生铁凋亡（铁依赖性的调节性坏死程序）的谷胱甘肽过氧化物酶4 （GPX4）在心肌梗死早期和中期下调。RNA-seq和qRT-PCR分析表明，GPX4下调发生在转录水平。使用特异性siRNA或化学抑制剂RSL3分别消耗或抑制GPX4，导致脂质过氧化积累，导致H9c2心肌细胞因铁下沉而死亡。尽管新生大鼠心室肌细胞（NRVMs）对GPX4抑制的敏感性低于H9c2细胞，但在半胱氨酸剥夺的情况下，NRVMs对GPX4抑制的反应迅速发生铁下垂。我们的研究表明，心肌梗死期间GPX4的下调有助于心肌细胞在代谢应激（如半胱氨酸剥夺）下的嗜铁细胞死亡。
Ischaemic heart disease (IHD) is the leading cause of death worldwide. Although myocardial cell death plays a significant role in myocardial infarction (MI), its underlying mechanism remains to be elucidated. To understand the progression of MI and identify potential therapeutic targets, we performed tandem mass tag (TMT)-based quantitative proteomic analysis using an MI mouse model. Gene ontology (GO) analysis and gene set enrichment analysis (GSEA) revealed that the glutathione metabolic pathway and reactive oxygen species (ROS) pathway were significantly downregulated during MI. In particular, glutathione peroxidase 4 (GPX4), which protects cells from ferroptosis (an iron-dependent programme of regulated necrosis), was downregulated in the early and middle stages of MI. RNA-seq and qRT-PCR analyses suggested that GPX4 downregulation occurred at the transcriptional level. Depletion or inhibition of GPX4 using specific siRNA or the chemical inhibitor RSL3, respectively, resulted in the accumulation of lipid peroxide, leading to cell death by ferroptosis in H9c2 cardiomyoblasts. Although neonatal rat ventricular myocytes (NRVMs) were less sensitive to GPX4 inhibition than H9c2 cells, NRVMs rapidly underwent ferroptosis in response to GPX4 inhibition under cysteine deprivation. Our study suggests that downregulation of GPX4 during MI contributes to ferroptotic cell death in cardiomyocytes upon metabolic stress such as cysteine deprivation.