title: Role of Nongenomic Signaling Pathways Activated by Aldosterone During Cardiac Reperfusion Injury.
abstract: Aldosterone (Aldo) activates both genomic and nongenomic signaling pathways in the cardiovascular system. Activation of genomic signaling pathways contributes to the adverse cardiac actions of Aldo during reperfusion injury; however, the extent nongenomic signaling pathways contribute has been difficult to identify due to lack of a specific ligand that activates only nongenomic signaling pathways. Using a pegylated aldosterone analog, aldosterone-3-carboxymethoxylamine-TFP ester conjugated to methoxypegylated amine (Aldo-PEG), we are able for the first time to distinguish between nongenomic and genomic cardiac actions of Aldo. We confirm Aldo-PEG activates phosphorylation of ERK1/2 in rat cardiomyocyte H9c2 cells similar to Aldo and G protein-coupled receptor 30 (GPR30 or GPER) agonist G1. GPER antagonist, G36, but not mineralocorticoid receptor (MR) antagonist spironolactone, prevented ERK1/2 phosphorylation by Aldo, Aldo-PEG, and G1. The selective nongenomic actions of Aldo-PEG are confirmed, with Aldo-PEG increasing superoxide production in H9c2 cells to similar levels as Aldo but having no effect on subcellular localization of MR. Striatin serves as a scaffold for GPER and MR, with GPER antagonist G36, but not spironolactone, restoring MR-striatin complexes. Aldo-PEG had no effect on MR-dependent transcriptional activation, whereas Aldo increased transcript levels of serum-regulated kinase 1 and plasminogen activator inhibitor-1. Using our ex vivo experimental rat model of myocardial infarction, we found aggravated infarct size and apoptosis by Aldo but not Aldo-PEG. Our studies confirm that in the heart, activation of nongenomic signaling pathways alone are not sufficient to trigger the deleterious effects of aldosterone during myocardial reperfusion injury.
authors.affiliation: Cardiovascular and Hormonal Research Laboratory, Department of Cardiology (T.Y.L.L., A.S.M.), Division of Perinatal Research (A.W.A.), Northern Blood Research Centre and Department of Haematology and Transfusion Medicine (M.-C.M.-K.), Sydney Neuro-Oncology Group and Bill Walsh Translational Cancer Research Laboratory (A.H.), Royal North Shore Hospital and Kolling Institute (A.W.A., T.Y.L.L., M.-C.M.-K., A.H., A.S.M.), Royal North Shore Hospital and The University of Sydney, Sydney 2065, Australia; Division of Endocrinology, G. V. (Sonny) Montgomery Veterans Affairs Medical Center and University of Mississippi Medical Center (C.E.G.-S.), Jackson, Mississippi 39216; and Analytical Chemistry Unit (B.M.), Pathology Queensland, Health Services Support Agency, Royal Brisbane and Women's Hospital, Brisbane, Queensland 4029, Australia.
authors.forename: Anthony W
authors.initials: AW
authors.lastname: Ashton
authors.affiliation: Cardiovascular and Hormonal Research Laboratory, Department of Cardiology (T.Y.L.L., A.S.M.), Division of Perinatal Research (A.W.A.), Northern Blood Research Centre and Department of Haematology and Transfusion Medicine (M.-C.M.-K.), Sydney Neuro-Oncology Group and Bill Walsh Translational Cancer Research Laboratory (A.H.), Royal North Shore Hospital and Kolling Institute (A.W.A., T.Y.L.L., M.-C.M.-K., A.H., A.S.M.), Royal North Shore Hospital and The University of Sydney, Sydney 2065, Australia; Division of Endocrinology, G. V. (Sonny) Montgomery Veterans Affairs Medical Center and University of Mississippi Medical Center (C.E.G.-S.), Jackson, Mississippi 39216; and Analytical Chemistry Unit (B.M.), Pathology Queensland, Health Services Support Agency, Royal Brisbane and Women's Hospital, Brisbane, Queensland 4029, Australia.
authors.forename: Thi Y L
authors.initials: TY
authors.lastname: Le
authors.affiliation: Cardiovascular and Hormonal Research Laboratory, Department of Cardiology (T.Y.L.L., A.S.M.), Division of Perinatal Research (A.W.A.), Northern Blood Research Centre and Department of Haematology and Transfusion Medicine (M.-C.M.-K.), Sydney Neuro-Oncology Group and Bill Walsh Translational Cancer Research Laboratory (A.H.), Royal North Shore Hospital and Kolling Institute (A.W.A., T.Y.L.L., M.-C.M.-K., A.H., A.S.M.), Royal North Shore Hospital and The University of Sydney, Sydney 2065, Australia; Division of Endocrinology, G. V. (Sonny) Montgomery Veterans Affairs Medical Center and University of Mississippi Medical Center (C.E.G.-S.), Jackson, Mississippi 39216; and Analytical Chemistry Unit (B.M.), Pathology Queensland, Health Services Support Agency, Royal Brisbane and Women's Hospital, Brisbane, Queensland 4029, Australia.
authors.forename: Celso E
authors.initials: CE
authors.lastname: Gomez-Sanchez
authors.affiliation: Cardiovascular and Hormonal Research Laboratory, Department of Cardiology (T.Y.L.L., A.S.M.), Division of Perinatal Research (A.W.A.), Northern Blood Research Centre and Department of Haematology and Transfusion Medicine (M.-C.M.-K.), Sydney Neuro-Oncology Group and Bill Walsh Translational Cancer Research Laboratory (A.H.), Royal North Shore Hospital and Kolling Institute (A.W.A., T.Y.L.L., M.-C.M.-K., A.H., A.S.M.), Royal North Shore Hospital and The University of Sydney, Sydney 2065, Australia; Division of Endocrinology, G. V. (Sonny) Montgomery Veterans Affairs Medical Center and University of Mississippi Medical Center (C.E.G.-S.), Jackson, Mississippi 39216; and Analytical Chemistry Unit (B.M.), Pathology Queensland, Health Services Support Agency, Royal Brisbane and Women's Hospital, Brisbane, Queensland 4029, Australia.
authors.forename: Marie-Christine
authors.initials: MC
authors.lastname: Morel-Kopp
authors.affiliation: Cardiovascular and Hormonal Research Laboratory, Department of Cardiology (T.Y.L.L., A.S.M.), Division of Perinatal Research (A.W.A.), Northern Blood Research Centre and Department of Haematology and Transfusion Medicine (M.-C.M.-K.), Sydney Neuro-Oncology Group and Bill Walsh Translational Cancer Research Laboratory (A.H.), Royal North Shore Hospital and Kolling Institute (A.W.A., T.Y.L.L., M.-C.M.-K., A.H., A.S.M.), Royal North Shore Hospital and The University of Sydney, Sydney 2065, Australia; Division of Endocrinology, G. V. (Sonny) Montgomery Veterans Affairs Medical Center and University of Mississippi Medical Center (C.E.G.-S.), Jackson, Mississippi 39216; and Analytical Chemistry Unit (B.M.), Pathology Queensland, Health Services Support Agency, Royal Brisbane and Women's Hospital, Brisbane, Queensland 4029, Australia.
authors.forename: Brett
authors.initials: B
authors.lastname: McWhinney
authors.affiliation: Cardiovascular and Hormonal Research Laboratory, Department of Cardiology (T.Y.L.L., A.S.M.), Division of Perinatal Research (A.W.A.), Northern Blood Research Centre and Department of Haematology and Transfusion Medicine (M.-C.M.-K.), Sydney Neuro-Oncology Group and Bill Walsh Translational Cancer Research Laboratory (A.H.), Royal North Shore Hospital and Kolling Institute (A.W.A., T.Y.L.L., M.-C.M.-K., A.H., A.S.M.), Royal North Shore Hospital and The University of Sydney, Sydney 2065, Australia; Division of Endocrinology, G. V. (Sonny) Montgomery Veterans Affairs Medical Center and University of Mississippi Medical Center (C.E.G.-S.), Jackson, Mississippi 39216; and Analytical Chemistry Unit (B.M.), Pathology Queensland, Health Services Support Agency, Royal Brisbane and Women's Hospital, Brisbane, Queensland 4029, Australia.
authors.forename: Amanda
authors.initials: A
authors.lastname: Hudson
authors.affiliation: Cardiovascular and Hormonal Research Laboratory, Department of Cardiology (T.Y.L.L., A.S.M.), Division of Perinatal Research (A.W.A.), Northern Blood Research Centre and Department of Haematology and Transfusion Medicine (M.-C.M.-K.), Sydney Neuro-Oncology Group and Bill Walsh Translational Cancer Research Laboratory (A.H.), Royal North Shore Hospital and Kolling Institute (A.W.A., T.Y.L.L., M.-C.M.-K., A.H., A.S.M.), Royal North Shore Hospital and The University of Sydney, Sydney 2065, Australia; Division of Endocrinology, G. V. (Sonny) Montgomery Veterans Affairs Medical Center and University of Mississippi Medical Center (C.E.G.-S.), Jackson, Mississippi 39216; and Analytical Chemistry Unit (B.M.), Pathology Queensland, Health Services Support Agency, Royal Brisbane and Women's Hospital, Brisbane, Queensland 4029, Australia.
authors.forename: Anastasia S
authors.initials: AS
authors.lastname: Mihailidou
mesh_terms: D000450:Aldosterone / Q000494:pharmacology*; D000588:Amines / Q000737:chemistry; D000818:Animals; D002148:Calmodulin-Binding Proteins / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D000451:Mineralocorticoid Receptor Antagonists / Q000494:pharmacology; D015428:Myocardial Reperfusion Injury; D009419:Nerve Tissue Proteins / Q000378:metabolism; D010766:Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D018161:Receptors, Mineralocorticoid / Q000378:metabolism; D015427:Reperfusion Injury / Q000378:metabolism*; D015398:Signal Transduction; D013148:Spironolactone / Q000494:pharmacology; D013481:Superoxides / Q000378:metabolism; D015533:Transcriptional Activation
chemical_list: D000588:Amines; D002148:Calmodulin-Binding Proteins; C520305:Gper1 protein, rat; D008565:Membrane Proteins; D000451:Mineralocorticoid Receptor Antagonists; D009419:Nerve Tissue Proteins; D043562:Receptors, G-Protein-Coupled; D018161:Receptors, Mineralocorticoid; C114095:Strn protein, rat; D013481:Superoxides; D013148:Spironolactone; D000450:Aldosterone