Liu Zhou, Chen Hongjin, Song Yingnan, Chen Kaiyuan, Pan Sisi, Yang Siyuan, Lu Deqin
School of Basic Medical Sciences, Guizhou Medical University, Guiyang, Guizhou, China.
Translational Medicine Research Center, Guizhou Medical University, Guiyang, Guizhou, China.
Front Pharmacol. 2024 Nov 13;15:1476718. doi: 10.3389/fphar.2024.1476718. eCollection 2024.
Myocardial ischemia-reperfusion injury (MIRI) is a prevalent complication in patients with myocardial infarction. The pathological mechanism of MIRI remains elusive. Ferroptosis plays a critical role in MIRI. This study aimed to investigate the role of spermidine/spermine N1-acetyltransferase 1 (Sat1) in MIRI by regulation of ferroptosis.
Rats and H9C2 cells were used to perform MIRI model. The extent of myocardial damage and associated pathological changes were evaluated. Protein expression was detected by western blot. Then we observed the mitochondrial morphology and measured cell viability and damage. The levels of lipid peroxide and glutathione were measured, and lipid reactive oxygen species (ROS) was quantified. Differentially expressed genes (DEGs) in MIRI were analyzed. Moreover, to explore the role of Sat1 in MIRI, this study utilized adeno-associated virus 9 and lentiviral transduction to modulate Sat1 expression in rats and H9C2 cells, respectively. The transcription factor that regulates Sat1 expression was predicated. Luciferase reporter gene experiment was conducted to reveal the potential sites of Sox2 binding to Sat1.
This study revealed that ferroptosis was involved in MIRI. Through bioinformatic analysis, Sat1 was identified as a significant gene in MIRI, which has been reported as an inducer of ferroptosis. Our results showed that Sat1 expression was significantly increased in MIRI. Next, the study showed that inhibition of Sat1 alleviated MIRI by suppressing ferroptosis in vivo and , and over-expression of Sat1 promoted MIRI via activation of ferroptosis. Furthermore, Sat1 and its interacting genes were enriched in several signaling pathways, including ferroptosis and the MAPK signaling pathway. The results showed that Sat1 regulated MIRI through ferroptosis via MAPK/ERK pathway. Moreover, it is found that Sox2 can suppress Sat1 expression at the transcriptional level. The potential binding site was TAACAAAGGAA.
In sum, this study demonstrated Sat1 expression was increased in MIRI, inhibition of Sat1 can alleviate MIRI by regulating ferroptosis via MAPK/ERK pathway, and Sat1 was negatively regulated by Sox2. These findings suggested that Sat1 may serve as a potential therapeutic target for the treatment of MIRI.
心肌缺血再灌注损伤（MIRI）是心肌梗死患者中常见的并发症。MIRI的病理机制仍不清楚。铁死亡在MIRI中起关键作用。本研究旨在通过调节铁死亡来探讨精胺/精脒N1 - 乙酰转移酶1（Sat1）在MIRI中的作用。
使用大鼠和H9C2细胞建立MIRI模型。评估心肌损伤程度及相关病理变化。通过蛋白质印迹法检测蛋白质表达。然后观察线粒体形态并测量细胞活力和损伤情况。测定脂质过氧化物和谷胱甘肽水平，并对脂质活性氧（ROS）进行定量分析。分析MIRI中差异表达基因（DEG）。此外，为了探究Sat1在MIRI中的作用，本研究分别利用腺相关病毒9和慢病毒转导来调节大鼠和H9C2细胞中Sat1的表达。预测调节Sat1表达的转录因子。进行荧光素酶报告基因实验以揭示Sox2与Sat1结合的潜在位点。
本研究表明铁死亡参与了MIRI。通过生物信息学分析，Sat1被确定为MIRI中的一个重要基因，据报道它是铁死亡的诱导剂。我们的结果显示Sat1在MIRI中表达显著增加。接下来，研究表明抑制Sat1可通过抑制体内铁死亡减轻MIRI，而Sat1的过表达则通过激活铁死亡促进MIRI。此外，Sat1及其相互作用基因富集于包括铁死亡和丝裂原活化蛋白激酶（MAPK）信号通路在内的多种信号通路中。结果表明Sat1通过MAPK/细胞外信号调节激酶（ERK）途径通过铁死亡调节MIRI。此外，发现Sox2可在转录水平抑制Sat1表达。潜在结合位点为TAACAAAGGAA。
总之，本研究表明Sat1在MIRI中表达增加，抑制Sat1可通过MAPK/ERK途径调节铁死亡来减轻MIRI，且Sat1受Sox2负调控。这些发现表明Sat1可能是治疗MIRI的潜在治疗靶点。
