Nitrones reverse hyperglycemia-induced endothelial dysfunction in bovine aortic endothelial cells

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• 作者：Colwyn A. Headley^a ; David DiSilvestro^d ; Kelsey E. Bryant^b ; ^c ; Craig Hemann^c ; Chun-An Chen^b ; ^c ; Amlan Das^a ; Ouliana Ziouzenkova^d ; Gré ; gory Durand^e ; Frederick A. Villamena^a ; ^b ; ^{villamena.1@osu.edu" class="auth_mail" title="E-mail the corresponding author}
• 关键词：AGEs ; advanced glycation end products ; LA ; alpha lipoic acid ; BAEC ; bovine aortic endothelial cells ; BCA ; bicinchoninic acid ; BH4 ; tetrahydrobiopterin ; CaI ; calcium ionophore ; CAT ; catalase (EC 1.11.1.6) ; CMD ; cardiometabolic disease ; CVD ; cardiovascular disease ; DAPI ; 4&prime ; 6-diamidino-2-phenylindole ; DHE ; dihydroethidium ; DMEM ; Dulbecco&rsquo ; s modified Eagle&rsquo ; s medium ; DTNB ; 5-(3-carboxy-4-nitrophenyl)disulfanyl-2-nitrobenzoic acid ; DTPA ; diethylene triamine penta-acetic acid ; ED ; end
• 刊名：Biochemical Pharmacology
• 出版年：2016
• 出版时间：15 March 2016
• 年：2016
• 卷：104
• 期：Complete
• 页码：108-117
• 全文大小：1313 K

文摘

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 (5.5 mM glucose, LG) or high glucose (50 mM, 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 24 h 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.