title: β3-Adrenoceptor activation relieves oxidative inhibition of the cardiac Na+-K+ pump in hyperglycemia induced by insulin receptor blockade.
abstract: Dysregulated nitric oxide (NO)- and superoxide (O2 (·-))-dependent signaling contributes to the pathobiology of diabetes-induced cardiovascular complications. We examined if stimulation of β3-adrenergic receptors (β3-ARs), coupled to endothelial NO synthase (eNOS) activation, relieves oxidative inhibition of eNOS and the Na(+)-K(+) pump induced by hyperglycemia. Hyperglycemia was established in male New Zealand White rabbits by infusion of the insulin receptor antagonist S961 for 7 days. Hyperglycemia increased tissue and blood indexes of oxidative stress. It induced glutathionylation of the Na(+)-K(+) pump β1-subunit in cardiac myocytes, an oxidative modification causing pump inhibition, and reduced the electrogenic pump current in voltage-clamped myocytes. Hyperglycemia also increased glutathionylation of eNOS, which causes its uncoupling, and increased coimmunoprecipitation of cytosolic p47(phox) and membranous p22(phox) NADPH oxidase subunits, consistent with NADPH oxidase activation. Blocking translocation of p47(phox) to p22(phox) with the gp91ds-tat peptide in cardiac myocytes ex vivo abolished the hyperglycemia-induced increase in glutathionylation of the Na(+)-K(+) pump β1-subunit and decrease in pump current. In vivo treatment with the β3-AR agonist CL316243 for 3 days eliminated the increase in indexes of oxidative stress, decreased coimmunoprecipitation of p22(phox) with p47(phox), abolished the hyperglycemia-induced increase in glutathionylation of eNOS and the Na(+)-K(+) pump β1-subunit, and abolished the decrease in pump current. CL316243 also increased coimmunoprecipitation of glutaredoxin-1 with the Na(+)-K(+) pump β1-subunit, which may reflect facilitation of deglutathionylation. In vivo β3-AR activation relieves oxidative inhibition of key cardiac myocyte proteins in hyperglycemia and may be effective in targeting the deleterious cardiac effects of diabetes.
authors.affiliation: North Shore Heart Research Group, Kolling Institute, University of Sydney, Sydney, Australia; and Department of Cardiology, Royal North Shore Hospital, Sydney, Australia.
authors.forename: Keyvan
authors.initials: K
authors.lastname: Karimi Galougahi
authors.affiliation: North Shore Heart Research Group, Kolling Institute, University of Sydney, Sydney, Australia; and.
authors.forename: Chia-Chi
authors.initials: CC
authors.lastname: Liu
authors.affiliation: North Shore Heart Research Group, Kolling Institute, University of Sydney, Sydney, Australia; and.
authors.forename: Alvaro
authors.initials: A
authors.lastname: Garcia
authors.affiliation: North Shore Heart Research Group, Kolling Institute, University of Sydney, Sydney, Australia; and.
authors.forename: Natasha A
authors.initials: NA
authors.lastname: Fry
authors.affiliation: North Shore Heart Research Group, Kolling Institute, University of Sydney, Sydney, Australia; and.
authors.forename: Elisha J
authors.initials: EJ
authors.lastname: Hamilton
authors.affiliation: North Shore Heart Research Group, Kolling Institute, University of Sydney, Sydney, Australia; and Department of Cardiology, Royal North Shore Hospital, Sydney, Australia.
authors.forename: Gemma A
authors.initials: GA
authors.lastname: Figtree
authors.affiliation: North Shore Heart Research Group, Kolling Institute, University of Sydney, Sydney, Australia; and Department of Cardiology, Royal North Shore Hospital, Sydney, Australia helge.rasmussen@sydney.edu.au.
authors.forename: Helge H
authors.initials: HH
authors.lastname: Rasmussen
mesh_terms: D058667:Adrenergic beta-3 Receptor Agonists / Q000494:pharmacology / Q000627:therapeutic use*; D000595:Amino Acid Sequence; D000818:Animals; D001786:Blood Glucose / Q000187:drug effects / Q000378:metabolism; D004149:Dioxoles / Q000494:pharmacology / Q000627:therapeutic use; D006943:Hyperglycemia / Q000188:drug therapy / Q000378:metabolism*; D008297:Male; D008969:Molecular Sequence Data; D018384:Oxidative Stress / Q000187:drug effects* / Q000502:physiology; D011817:Rabbits; D011972:Receptor, Insulin / Q000037:antagonists & inhibitors*; D022702:Receptors, Adrenergic, beta-3 / Q000378:metabolism*; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors*
chemical_list: D058667:Adrenergic beta-3 Receptor Agonists; D001786:Blood Glucose; D004149:Dioxoles; D022702:Receptors, Adrenergic, beta-3; C076126:disodium (R,R)-5-(2-((2-(3-chlorophenyl)-2-hydroxyethyl)-amino)propyl)-1,3-benzodioxole-2,3-dicarboxylate; D011972:Receptor, Insulin; D000254:Sodium-Potassium-Exchanging ATPase
keywords: Na+-K+ pump; diabetes; endothelial nitric oxide synthase; glutathionylation; hyperglycemia; β3-adrenergic receptors