Department of Forensic Medicine, School of Basic Medical Sciences, Fudan University, Shanghai, 200032, China; Department of Forensic Medicine, School of Basic Medical Sciences, Gannan Medical University, Ganzhou, 341000, China.
Department of Forensic Medicine, School of Basic Medical Sciences, Fudan University, Shanghai, 200032, China.
Exp Cell Res. 2024 May 15;438(2):114061. doi: 10.1016/j.yexcr.2024.114061. Epub 2024 Apr 30.
Acute myocardial infarction (AMI) is a prevalent cardiovascular disease with high morbidity and mortality rates worldwide. Pyroptosis is an inflammatory form of programmed cell death that has been linked to various pathological conditions. However, its exact contribution to the onset and progression of heart injury in AMI has not yet fully elucidated. Herein, we established mouse AMI model by ligating the left anterior descending artery and performed transcriptome analysis during the early phase of AMI. Mouse HL-1 and human AC-16 cardiomyocytes were subjected to hypoxia to simulate ischemic injury in vitro. Our results revealed a significant activation of the inflammatory response at 3 h post-ligation, as confirmed by RNA sequencing. We identified the occurrence of NLRP3 inflammasome-mediated pyroptosis in the cardiac tissues of human cases with AMI, as well as in mouse models of AMI and hypoxia-induced cardiomyocytes, using immunohistochemistry staining and Western blotting assays. Concurrently, pharmacological inhibition of NLRP3 inflammasome-mediated pyroptosis with MCC950 and VX-765 effectively decreased hypoxia-induced cardiomyocytes injury, while mitigating myocardial oxidative stress, apoptosis and inflammation caused by hypoxia. Moreover, the circulating levels of gasdermin D (GSDMD), the pyroptosis executor, were remarkably elevated in the plasma of mice with early AMI and in the supernatant of hypoxia-exposed cardiomyocytes in a time-dependent manner using ELISA and Western blotting. Furthermore, the change in circulating GSDMD positively correlated with Creatine Kinase-MB (CK-MB) in the plasma of early-stage AMI mouse. In summary, these findings indicated a critical role for NLRP3 inflammasome-mediated pyroptosis in the progression of AMI, the administration of MCC950 and VX-765 may be attractive candidate therapeutic approaches for cardiac injury caused by acute hypoxia or even AMI. Additionally, the circulating GSDMD exhibits potential as a newly diagnostic biomarker for AMI.
急性心肌梗死（AMI）是一种普遍存在的心血管疾病，具有高发病率和死亡率。细胞焦亡是一种炎症形式的程序性细胞死亡，与各种病理状态有关。然而，它对 AMI 中心脏损伤的发生和发展的确切贡献尚未完全阐明。在此，我们通过结扎左前降支建立了小鼠 AMI 模型，并在 AMI 的早期阶段进行了转录组分析。体外将小鼠 HL-1 和人 AC-16 心肌细胞置于缺氧条件下模拟缺血性损伤。我们的结果显示，在结扎后 3 小时，通过 RNA 测序证实炎症反应明显激活。我们使用免疫组织化学染色和 Western blot 检测发现，在 AMI 患者的心脏组织以及 AMI 小鼠模型和缺氧诱导的心肌细胞中，存在 NLRP3 炎性体介导的细胞焦亡。同时，使用 MCC950 和 VX-765 抑制 NLRP3 炎性体介导的细胞焦亡可有效减轻缺氧诱导的心肌细胞损伤，减轻缺氧引起的心肌氧化应激、凋亡和炎症。此外，ELISA 和 Western blot 检测结果显示，在早期 AMI 小鼠的血浆中和缺氧暴露的心肌细胞上清液中，气体渗透酶 D（GSDMD）的执行器焦亡的循环水平呈时间依赖性显著升高。此外，早期 AMI 小鼠血浆中循环 GSDMD 的变化与血浆中肌酸激酶同工酶-MB（CK-MB）呈正相关。综上所述，这些发现表明 NLRP3 炎性体介导的细胞焦亡在 AMI 的进展中起关键作用，MCC950 和 VX-765 的给药可能是急性缺氧甚至 AMI 引起的心脏损伤的有吸引力的治疗方法。此外，循环 GSDMD 有望成为 AMI 的新诊断生物标志物。
