Knocking down NON796 inhibits ferroptosis and alleviates myocardial ischemia-reperfusion injury in mice by targeting the miR-486a-5p/Furin axis

Abstract Objective: Investigating the roles of NON796, miR-486a-5p, and Furin in controlling myocardial ischemia/reperfusion (MI/R) damage and their possible therapeutic applications in cell function and iron death. Methods: We adopted bioinformatics methods and conducted a detailed analysis of dataset GSE161151 in the Gene Expression Omnibus (GEO) to deeply understand the molecular mechanism of myocardial ischemia-reperfusion (MI/R) injury. Bioinformatics identified miR-486a-5p as a downstream target of NON796, with binding sites confirmed through RT-qPCR, dual-luciferase assays, and gene expression analysis. In a hypoxia/reoxygenation (H/R) model, the effects of NON796 knockdown and miR-486a-5p inhibition were evaluated by measuring LDH release, CK-MB, Fe2+ levels, and cell viability. Protein levels of iron metabolism-related genes (Tfrc, Fth1, Ftl1, Gpx4, Furin) were assessed by PCR and Western blot. A mouse MI/R model was used to analyze myocardial injury via TTC staining and measure serum LDH and CK-MB levels, as well as the expression of NON796, miR-486a-5p, and Furin in heart tissue. Results: In the mouse ischemic model, NON796 was upregulated, and its knockdown or miR-486a-5p overexpression reversed metabolic dysfunction caused by H/R injury, suggesting NON796 negatively regulates miR-486a-5p. NON796 knockdown reduced LDH release and improved cell viability, while miR-486a-5p inhibition reversed these effects. Furin was shown to be a target of miR-486a-5p by bioinformatics, and the effects of miR-486a-5p were reversed by Furin overexpression. In vivo, NON796 was upregulated, miR-486a-5p downregulated, and Furin elevated in the infarct zone of the MI/R model, with increased iron accumulation in the ischemic myocardium, indicating the role of iron death in myocardial injury. Conclusion: NON796 regulates MI/R injury by controlling miR-486a-5p, which impacts iron death and cell function. Furin, a key target of miR-486a-5p, mediates these effects. Targeting the NON796/miR-486a-5p/Furin axis may offer a promising therapeutic approach for MI/R injury.