Cellular Physiology and Biochemistry · Published 2025-12-31 · DOI 10.33594/000000837
Background/Aims: Myocardial infarction (MI) remains the leading cause of cardiovascular morbidity and mortality. Although reperfusion therapy restores myocardial blood flow, it also induces myocardial ischemia-reperfusion injury (MI/RI), which worsens prognosis. Ferroptosis, an iron-dependent form of regulated cell death driven by excessive lipid reactive oxygen species (ROS), plays a central role in MI/RI. This process is characterized by a downregulation of glutathione peroxidase 4 (GPX4) and an upregulation of acyl-CoA synthetase long-chain family member 4 (ACSL4). While long non-coding RNA (lncRNA) ANRIL has been found to be aberrantly expressed in acute myocardial infarction (AMI) and is believed to provide myocardial protection, its role and underlying mechanism in MI/RI-induced ferroptosis remain unclear. Methods: A mouse MI/R model was established by ligating the left anterior descending (LAD) coronary artery in C57BL/6 mice. In parallel, HL-1 and H9C2 cardiomyocytes underwent hypoxia-reoxygenation (H/R) to simulate MI/RI in vitro. Lnc-ANRIL was either overexpressed using the pEGFP-lnc-ANRIL plasmid or silenced with an siRNA targeting ANRIL (siANRIL). Ferroptosis indicators, including ROS, malondialdehyde (MDA), Fe²⁺, GPX4, and ACSL4, were assessed. Potential microRNAs (miRNAs) targeting lnc-ANRIL and GPX4 were predicted by microRNA Target Prediction Database (miRDB) and validated through dual-luciferase assays. Results: In both MI/R mouse myocardial tissue and H/R-treated cardiomyocytes, ferroptosis was activated, as evidenced by downregulated GPX4, upregulated ACSL4, and increased levels of ROS, MDA, and Fe²⁺. Concurrently, lnc-ANRIL expression was reduced. Overexpression of lnc-ANRIL attenuated H/R-induced ferroptosis, reduced cell death and ferroptosis markers, and upregulated GPX4 expression. Conversely, silencing lnc-ANRIL exacerbated ferroptosis. Mechanistically, lnc-ANRIL acted as a sponge for miR-7238-3p, which targets the 3'-UTR of GPX4 to suppress its expression. Overexpression of miR-7238-3p mimicked the ferroptosis phenotype associated with low lnc-ANRIL expression and exacerbated cardiomyocyte damage. Conclusion: MI/RI downregulates lnc-ANRIL, thereby relieving the inhibition of miR-7238-3p. This, in turn, suppresses GPX4 expression and triggers ferroptosis in cardiomyocytes. Lnc-ANRIL protects against MI/RI-induced ferroptosis through the miR-7238-3p/GPX4 axis, identifying a potential novel therapeutic target for ischemic heart disease.
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