Aldo-Keto reductase 1C3 reduces myocardial cell damage after acute myocardial infarction by activating the Kelch-like ECH-associated protein 1-nuclear factor erythroid 2-related factor 2-antioxidant response element pathway to inhibit ferroptosis

Background Acute myocardial infarction(AMI)is a high-risk cardiovascular condition associated with increased cellular damage and oxidative stress.Aldo-Keto Reductase 1C3(AKR1C3)is a stress-regulating gene.Nevertheless,its specific role and mechanisms regarding AMI remain unclear.Methods We assessed cardiac function through echocardiography;tissue damage was evaluated using Hematoxylin and Eosin(HE)and Masson trichrome staining.AKR1C3 expression levels were measured through Reverse transcription-quantitative polymerase chain reaction and western blot.Assessed cell viability using Cell Counting Kit-8 and lactate dehydrogenase(LDH)assays.The extent of ferroptosis was determined by measuring the levels of Fe2+,boron-dipyrromethane(BODIPY)and malondialdehyde(MDA),the glutathione/glutathione disulfide(GSH/GSSG)ratio,and the expression of Glutathione Peroxidase 4(GPX4)and Solute carrier 7A11(SLC7A11).Kelch-like ECH-associated protein 1-Nuclear factor erythroid 2-related factor 2-Antioxidant response element(Keap1-Nrf2-ARE)pathway activation was analyzed through western blotting.Nrf2 was inhibited with ML385and activated with(R)-Sulforaphane to investigate the Keap1-Nrf2-ARE pathway.Results The rats in the AMI group displayed reduced heart function,more tissue damage,and lower AKR1C3 expression compared to the Sham group.Similarly,hypoxia-treated H9C2 cells showed reduced viability,and decreased AKR1C3 expression.Overexpressing AKR1C3 in H9C2 cells enhanced viability.Knocking down AKR1C3 exhibited the opposite effect.Of the inhibitors tested,Ferrostatin-1 most effectively restored cell viability in hypoxia-treated H9C2 cells.Moreover,H9C2 cells subjected to hypoxia suggested Keap1-Nrf2-ARE pathway inhibition.Overexpressing AKR1C3 reduced ferroptosis and activated the Keap1-Nrf2-ARE pathway in hypoxia-treated cells,knocking down AKR1C3 exhibited the opposite effect.Further experiments using ML385 in hypoxia-treated H9C2 cells with overexpressed AKR1C3 showed decreased viability and increased ferroptosis compared to the control.Using(R)-Sulforaphane in hypoxia-treated H9C2 cells with knocked-down AKR1C3 exhibited the opposite effect.Conclusion This study's findings indicate that AKR1C3 plays a role in regulating ferroptosis in myocardial cells,with the Keap1-Nrf2-ARE pathway likely being a key mechanism behind it.