Wang Deliang, Dai Zheng, Jiang Lu, Liu Ke
Department of Cardiac Surgery, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, Chengdu, Sichuan, 610072, China.
Department of Emergency Medicine, Sichuan Provincial People's Hospital, University of Electronic Science and Technology of China, Chengdu, Sichuan, 610072, China.
Biol Direct. 2025 Mar 26;20(1):36. doi: 10.1186/s13062-025-00605-2.
In the quest to elucidate novel therapeutic strategies for myocardial injury, recent investigations have underscored the pivotal roles played by circulating extracellular vesicles (EVs) in intercellular communication.
EVs were extracted from individuals who had experienced AMI-EVs and those who were N-EVs. To assess the impact of circulating EVs on cardiomyocyte and endothelial cell proliferation, apoptosis, migration, and tube formation, a range of in vitro assays such as CCK8, EdU assays, flow cytometry, wound healing assays and angiogenesis assays were conducted. Differentially expressed miRNAs in EVs were validated using microarray analysis and real-time PCR. Through bioinformatics analysis, ELAVL1 was identified as a potential downstream target of miR-133a-3p. This finding was further confirmed by conducting dual-luciferase reporter assay and RNA co-immunoprecipitation experiments. To investigate the regulatory effects of circulating EVs from various sources on myocardial injury and PANoptosis, an animal model of ischemia-reperfusion-induced myocardial injury was established.
Our findings revealed that circulating EVs effectively deliver miR-133a-3p to target cells, where it binds to ELAVL1, leading to a decrease in NLRP3 mRNA stability. This reduction in NLRP3 mRNA stability subsequently inhibits the assembly of the PANoptosome, a multi-protein complex implicated in PANoptosis. As a result, we observed a significant mitigation of PANoptosis in our myocardial injury models, demonstrating the protective role of miR-133a-3p against excessive cell death.
The present study underscores the regulatory role of circulating EV-delivered miR-133a-3p in modulating PANoptosis through ELAVL1-mediated NLRP3 mRNA stabilization. This mechanism represents a potential therapeutic target for attenuating myocardial injury by suppressing PANoptosis.
在寻求阐明心肌损伤新治疗策略的过程中，最近的研究强调了循环细胞外囊泡（EVs）在细胞间通讯中所起的关键作用。
从经历过急性心肌梗死的个体（AMI-EVs）和未经历过的个体（N-EVs）中提取EVs。为了评估循环EVs对心肌细胞和内皮细胞增殖、凋亡、迁移和管腔形成的影响，进行了一系列体外实验，如CCK8、EdU实验、流式细胞术、伤口愈合实验和血管生成实验。使用微阵列分析和实时PCR验证EVs中差异表达的miRNA。通过生物信息学分析，确定ELAVL1是miR-133a-3p的潜在下游靶点。通过进行双荧光素酶报告基因实验和RNA免疫共沉淀实验进一步证实了这一发现。为了研究来自不同来源的循环EVs对心肌损伤和PANoptosis的调节作用，建立了缺血再灌注诱导的心肌损伤动物模型。
我们的研究结果表明，循环EVs有效地将miR-133a-3p传递到靶细胞，在那里它与ELAVL1结合，导致NLRP3 mRNA稳定性降低。NLRP3 mRNA稳定性的这种降低随后抑制了PANoptosome的组装，PANoptosome是一种与PANoptosis有关的多蛋白复合物。结果，我们在心肌损伤模型中观察到PANoptosis显著减轻，证明了miR-133a-3p对过度细胞死亡的保护作用。
本研究强调了循环EV传递的miR-133a-3p通过ELAVL1介导的NLRP3 mRNA稳定在调节PANoptosis中的作用。这一机制代表了通过抑制PANoptosis减轻心肌损伤的潜在治疗靶点。
