Nitrones reverse hyperglycemia-induced endothelial dysfunction in bovine aortic endothelial cells.
Nitrones reverse hyperglycemia-induced endothelial dysfunction in bovine aortic endothelial cells.
复制标题
硝基反向高血糖诱导的牛主动脉内皮细胞中的内皮功能障碍。
DOI：
10.1016/j.bcp.2016.01.005
复制
发表时间：
2016-03-15
影响因子：
5.8
通讯作者：
Villamena FA
中科院分区：
文献类型：
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作者：
Headley CA;DiSilvestro D;Bryant KE;Hemann C;Chen CA;Das A;Ziouzenkova O;Durand G;Villamena FA
高血糖通过增加ROS的产生与内皮功能障碍的发生有关。由于硝酮可逆转eNOS功能障碍，提高抗氧化酶活性，并抑制ROS诱导的促凋亡信号通路和线粒体功能障碍，本研究的目的是在体外利用牛主动脉内皮细胞(BAEC)建立高血糖诱导的功能障碍模型，以确定硝酮自旋陷阱DMPO、PBN和PBN-LA是否能有效复制这些作用并改善葡萄糖摄取。将BAEC在含有低糖(5.5 mM，LG)和高糖(50 mM，HG)的DMEM培养液中培养14天，建立代谢性疾病患者体内高血糖模型。硝酮单独作用BAEC细胞24 h后，细胞存活率、细胞内氧化应激、NO和四氢生物蝶呤水平、线粒体膜电位、葡萄糖转运和抗氧化酶活性均有改善。慢性高血糖使细胞内ROS显著增加50%，细胞存活率降低25%，NO生物利用度降低50%，BH4水平降低15%，从而减少NO的产生。细胞内葡萄糖转运和超氧化物歧化酶活性分别降低50%和25%。硝酮(PBN和DMPO，50μM)处理在高血糖条件下生长的BAEC细胞后，除超氧化物歧化酶和过氧化氢酶活性外，其余指标均恢复正常。我们的发现表明，硝酮逆转了高血糖对BAEC细胞的有害影响。我们认为，在心脏代谢性疾病模型中对这些硝酮化合物进行体内测试是有必要的。
Hyperglycemia has been implicated in the development of endothelial dysfunction through heightened ROS production. Since nitrones reverse 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 levels, mitochondrial membrane potential, glucose transport, and activity of antioxidant enzymes were measured from single treatment of BAEC cells 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 SOD activity were also decreased by 50% and 25% respectively. Nitrone (PBN and DMPO, 50 μM) treatment of BAEC cells grown in hyperglycemic conditions resulted in 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 cells. We believe that in vivo testing of these nitrone compounds in models of cardiometabolic disease is warranted.