Analysis of cardiovascular complications by the production of reactive oxygen species from mitochondria in patients with glucocrticoid excess

在cultured human vascular endothelial cells from umbilical vecord in（HUVEO）中使用荧光探针（CM-H-H-D22 DCFDA、DHR 123和DAF-2）中使用荧光探针（CM-H-H-D22 DCFDA、DHR 123和DAF-2）进行研究。ROS和过氧化物酶的产物明显地增加了，但NO的产物明显地减少了10-D1-7-D1 M地塞米松处理的HUVEC。对3-硝基酪氨酸氨酸的西方印迹分析（过氧化物酶的脚印）显示了在脱氢酶处理HUVEC中过氧化物酶的增加。过量生产由葡萄糖皮质激素引起的过量生产被碳基氰化物M-氯苯胺肼市场上的过量生产，可以从三氯酮反应链中降解ROS生产。Furthermore，对复方I的抑制剂通过氯二苯碘嗪，对复方11的抑制剂通过氟三氟代酮明显地降解在二甲苯甲酮处理的HLVEC中过度产生ROS。However，对复合体111的抑制不能通过myxothiazol来消除经修饰的HUVEC处理的ROS过度生产。这些发现揭示了葡萄糖eorticoid可能导致复合体I和II中的ROS过度生产，以及血管内端细胞中NO的间接消费。ROS的产生和NO的消费由葡萄糖皮质激素例外可能会影响血管内端功能，因为它可能是患者与葡萄糖皮质激素例外的心血管复杂性的主要病理学。

The effect of glucbcrticoid excess on the production of reactive oxygen species (ROS), peroxynitrate and nitric oxide (NO) was investigated using the fluorescence probe (CM-HィイD22ィエD2DCFDA, DHR123 and DAF-2) in cultured human vascular endothelial cells from umbilical cord vein (HUVEO). The productions of ROS and peroxynitrate were significantly increased, but the production of NO was significantly decreased in 10ィイD1-7ィエD1M dexamethasone-treated HUVEC. Western blot analysis of 3-nitrotyrosine (foot print of peroxynitrate) showed increase of peroxynitrate in dexamethasone-treated HUVEC. Over-production by glucocorticoid excess was markedly suppressed by carbonyl cyanide m-chlorophenylhyazone, which could decrease ROS production from mitochondrial respiratory chain. Furthermore, the inhibition of complex I by diphenyleneiodinium chloride and the inhibition of complex 11 by thenoyltrifluoroadetone significantly decrease the over-production of ROS in dexamethasone-treated HLVEC. However, the inhibition of complex 111 by myxothiazol could not decrease the over-production of ROS in dexamethasone-treated HUVEC.These findings showed that glucoeorticoid excess could cause the over-production of ROS from complex I and II in mitochondrial respiratory chain, and subsequently the consumption of NO in vascular endothekial cells. The production of ROS and consumption of NO by glucocorticoid excess could impair the vascular endothelial function, which may be a major pathogenesis for cardiovascular complications in patients with glucocorticoid excess.