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Updated: Aug 7, 2026
Improved Rodent Model of Myocardial Ischemia and Reperfusion Injury
Published on: March 7, 2022
Empagliflozin Attenuates Ferroptosis in Acute Myocardial Infarction via the AMPK/NRF2/SLC7A11 Signaling Pathway
Xiangrui Tang1, Xiaoxia Zhao2, Chunjun Liu3
1Department of Cardiology, 3201 Hospital of Xi'an Jiaotong University Health Science Center.
Abstract:
Emerging evidence implicates ferroptosis in myocardial ischemic injury. This study aimed to investigate whether empagliflozin (EMP) suppresses ferroptosis in acute myocardial infarction (AMI) via the AMP-activated protein kinase (AMPK)/nuclear factor erythroid 2-related factor 2 (NRF2)/solute carrier family 7 member 11 (SLC7A11) pathway.Hypoxia/reoxygenation (H/R)-injured HL-1 cardiomyocytes were treated with EMP (10, 20, 30 μM). Cell viability, morphology, damage, and apoptosis were assessed by CCK-8, inverted microscopy, lactate dehydrogenase (LDH) release, and flow cytometry. H/R-injured cells were treated with 30 μM EMP and/or ferrostatin-1 (Fer-1), followed by measurements of Fe2+, malondialdehyde (MDA), glutathione (GSH), reactive oxygen species (ROS), superoxide dismutase (SOD), lipid ROS (C11-BODIPY), ferroptosis-related proteins [SLC7A11, glutathione peroxidase 4 (GPX4), phospho-AMPK (p-AMPK), AMPK, NRF2 using Western blot], and NRF2 nuclear translocation (using immunofluorescence). An AMI mouse model was established. Myocardial pathology and infarct size were evaluated, and AMPK/NRF2/SLC7A11 pathway proteins and myocardial Fe2+/MDA/GSH levels were measured.EMP partially reversed H/R-induced cardiomyocyte shrinkage and membrane rupture, reduced viability, elevated LDH, and increased apoptosis. Among cell death inhibitors, Fer-1 exerted maximal protection, indicating ferroptosis may be the predominant death type in H/R-injured HL-1 cardiomyocytes. EMP mirrored the anti-ferroptotic activity of Fer-1. Mechanistically, EMP decreased cytoplasmic NRF2 and the p-AMPK/AMPK ratio while promoting the translocation of NRF2 from the cytoplasm to the nucleus. EMP activated the AMPK/NRF2/SLC7A11 axis to attenuate ferroptosis in AMI mice.In conclusion, EMP suppressed ferroptosis and ameliorated cellular injury in H/R-injured cardiomyocytes by activating the AMPK/NRF2/SLC7A11 pathway, with efficacy confirmed in vivo during AMI.
Insights
Empagliflozin (EMP) mitigates ferroptosis in acute myocardial infarction (AMI) by activating the AMP-activated protein kinase (AMPK)/nuclear factor erythroid 2-related factor 2 (NRF2)/solute carrier family 7 member 11 (SLC7A11) pathway, protecting heart cells from injury.
Area of Science:
- Cardiology
- Molecular Biology
- Biochemistry
Background:
- Ferroptosis, a regulated form of cell death, is increasingly implicated in myocardial ischemic injury.
- Understanding the molecular mechanisms underlying ferroptosis in acute myocardial infarction (AMI) is crucial for developing effective therapeutic strategies.
Purpose of the Study:
- To investigate the potential of empagliflozin (EMP) in suppressing ferroptosis in AMI.
- To elucidate the role of the AMP-activated protein kinase (AMPK)/nuclear factor erythroid 2-related factor 2 (NRF2)/solute carrier family 7 member 11 (SLC7A11) pathway in EMP's protective effects.
Main Methods:
- HL-1 cardiomyocytes subjected to hypoxia/reoxygenation (H/R) injury were treated with EMP.
- Cell viability, damage markers (LDH), apoptosis, and ferroptosis indicators (Fe2+, MDA, GSH, lipid ROS) were measured.
- Western blot and immunofluorescence were used to assess protein expression and NRF2 nuclear translocation.
- An in vivo AMI mouse model was utilized to evaluate myocardial pathology and infarct size.
Main Results:
- EMP partially reversed H/R-induced cardiomyocyte damage and apoptosis, mirroring the protective effects of the ferroptosis inhibitor ferrostatin-1 (Fer-1).
- EMP treatment led to decreased cytoplasmic NRF2 and p-AMPK/AMPK ratio, while promoting NRF2 nuclear translocation.
- EMP activated the AMPK/NRF2/SLC7A11 axis, reducing ferroptosis markers in both in vitro and in vivo AMI models.
Conclusions:
- Empagliflozin effectively suppresses ferroptosis and ameliorates cellular injury in cardiomyocytes following H/R insult.
- The protective mechanism involves the activation of the AMPK/NRF2/SLC7A11 pathway.
- EMP demonstrates therapeutic potential for treating acute myocardial infarction by targeting ferroptosis.
