title: Nitrones reverse hyperglycemia-induced endothelial dysfunction in bovine aortic endothelial cells.
abstract: Hyperglycemia has been implicated in the development of endothelial dysfunction through heightened ROS production. Since nitrones reverse endothelial nitric oxide synthase (eNOS) dysfunction, increase antioxidant enzyme activity, and suppress pro-apoptotic signaling pathway and mitochondrial dysfunction from ROS-induced toxicity, the objective of this study was to determine whether nitrone spin traps DMPO, PBN and PBN-LA were effective at duplicating these effects and improving glucose uptake in an in vitro model of hyperglycemia-induced dysfunction using bovine aortic endothelial cells (BAEC). BAEC were cultured in DMEM medium with low (5.5mM glucose, LG) or high glucose (50mM, HG) for 14 days to model in vivo hyperglycemia as experienced in humans with metabolic disease. Improvements in cell viability, intracellular oxidative stress, NO and tetrahydrobiopterin (BH4)​ levels, mitochondrial membrane potential, glucose transport, and activity of antioxidant enzymes were measured from single treatment of BAEC with nitrones for 24h after hyperglycemia. Chronic hyperglycemia significantly increased intracellular ROS by 50%, decreased cell viability by 25%, reduced NO bioavailability by 50%, and decreased (BH4) levels by 15% thereby decreasing NO production. Intracellular glucose transport and superoxide dismutase (SOD) activity were also decreased by 50% and 25% respectively. Nitrone (PBN and DMPO, 50 μM) treatment of BAEC grown in hyperglycemic conditions resulted in the normalization of outcome measures except for SOD and catalase activities. Our findings demonstrate that the nitrones reverse the deleterious effects of hyperglycemia in BAEC. We believe that in vivo testing of these nitrone compounds in models of cardiometabolic disease is warranted.
authors.affiliation: Department of Biological Chemistry and Pharmacology, The Ohio State University, Columbus, OH, USA.
authors.forename: Colwyn A
authors.initials: CA
authors.lastname: Headley
authors.affiliation: Department of Human Nutrition, College of Education and Human Ecology, The Ohio State University, Columbus, OH, USA.
authors.forename: David
authors.initials: D
authors.lastname: DiSilvestro
authors.affiliation: Department of Emergency Medicine, The Ohio State University, Columbus, OH, USA; The Davis Heart and Lung Research Institute, College of Medicine, The Ohio State University, Columbus, OH, USA.
authors.forename: Kelsey E
authors.initials: KE
authors.lastname: Bryant
authors.affiliation: The Davis Heart and Lung Research Institute, College of Medicine, The Ohio State University, Columbus, OH, USA.
authors.forename: Craig
authors.initials: C
authors.lastname: Hemann
authors.affiliation: Department of Emergency Medicine, The Ohio State University, Columbus, OH, USA; The Davis Heart and Lung Research Institute, College of Medicine, The Ohio State University, Columbus, OH, USA.
authors.forename: Chun-An
authors.initials: CA
authors.lastname: Chen
authors.affiliation: Department of Biological Chemistry and Pharmacology, The Ohio State University, Columbus, OH, USA.
authors.forename: Amlan
authors.initials: A
authors.lastname: Das
authors.affiliation: Department of Human Nutrition, College of Education and Human Ecology, The Ohio State University, Columbus, OH, USA.
authors.forename: Ouliana
authors.initials: O
authors.lastname: Ziouzenkova
authors.affiliation: Institut des Biomolécules Max Mousseron (IBMM), UMR 5247, CNRS-Université Montpellier-ENSCM & Avignon Université, Equipe Chimie Bioorganique et Systèmes Amphiphiles, 33 rue Louis Pasteur, 84000 Avignon, France.
authors.forename: Grégory
authors.initials: G
authors.lastname: Durand
authors.affiliation: Department of Biological Chemistry and Pharmacology, The Ohio State University, Columbus, OH, USA; Department of Emergency Medicine, The Ohio State University, Columbus, OH, USA. Electronic address: villamena.1@osu.edu.
authors.forename: Frederick A
authors.initials: FA
authors.lastname: Villamena
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D001011:Aorta / Q000187:drug effects* / Q000378:metabolism / Q000503:physiopathology; D002417:Cattle; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000378:metabolism / Q000503:physiopathology; D006943:Hyperglycemia / Q000378:metabolism / Q000503:physiopathology*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D009569:Nitric Oxide / Q000378:metabolism; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D009589:Nitrogen Oxides / Q000494:pharmacology* / Q000633:toxicity; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D000975:Antioxidants; D009589:Nitrogen Oxides; D017382:Reactive Oxygen Species; C015388:nitrones; D009569:Nitric Oxide; D052250:Nitric Oxide Synthase Type III
keywords: 5,5-dimethyl-1-pyrroline N-oxide (PubChem ID: 1774); Endothelial dysfunction; Hyperglycemia; N-tert-butyl-α-phenylnitrone (PubChem ID: 638877); NO; ROS; α-lipoic acid (PubChem ID: 864)