Chronic Deficiency of Nitric Oxide Affects Hypoxia Inducible Factor-1α (HIF-1α) Stability and Migration in Human Endothelial Cells
Chronic Deficiency of Nitric Oxide Affects Hypoxia Inducible Factor-1α (HIF-1α) Stability and Migration in Human Endothelial Cells
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DOI：
10.1371/journal.pone.0029680
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发表时间：
2011-12-27
期刊：
影响因子：
3.7
通讯作者：
Vicentini, Lucia M.
中科院分区：
文献类型：
--
作者：
Cattaneo, Maria Grazia;Cappellini, Elisa;Vicentini, Lucia M.
背景资料：广泛弥漫性疾病如动脉粥样硬化、高血压、糖尿病和衰老中的内皮功能障碍与一氧化氮（NO）缺乏有关。在这里，来自人脐静脉内皮细胞（HUVECs）NO缺乏的行为和分子后果进行了调查。结果：内皮型一氧化氮合酶（eNOS）被慢性抑制无论是由N-G-硝基-L-精氨酸甲酯（L-NAME）治疗或其表达下调RNA干扰。长期L-NAME处理后，HUVEC显示出更高的迁移能力，伴随着血管内皮生长因子（VEGF）和VEGF受体-2（激酶插入结构域受体，KDR）表达的增加。此外，eNOS的药理学和遗传学抑制诱导了假性缺氧状态，这通过缺氧诱导因子-1 α（HIF-1 α）的稳定来揭示。此外，NO损失诱导线粒体质量和能量产生的显着下降，伴随着较低的O-2消耗。值得注意的是，非常低剂量的长期管理DETA/NO逆转HIF-1 α积累，增加VEGF表达和刺激迁移行为检测NO deficient cells.Conclusion：根据我们的研究结果，我们建议，基础释放NO可能作为一个负控制器HIF-1 α水平与内皮细胞生理学的重要后果。此外，我们认为，我们的实验模型，eNOS活性受损的药理学和遗传抑制可能是一个很好的体外系统来研究内皮功能障碍。
Background: Endothelial dysfunction in widely diffuse disorders, such as atherosclerosis, hypertension, diabetes and senescence, is associated with nitric oxide (NO) deficiency. Here, the behavioural and molecular consequences deriving from NO deficiency in human umbilical vein endothelial cells (HUVECs) were investigated.Results: Endothelial nitric oxide synthase (eNOS) was chronically inhibited either by N-G-Nitro-L-arginine methyl ester (L-NAME) treatment or its expression was down-regulated by RNA interference. After long-term L-NAME treatment, HUVECs displayed a higher migratory capability accompanied by an increased Vascular Endothelial Growth Factor (VEGF) and VEGF receptor-2 (kinase insert domain receptor, KDR) expression. Moreover, both pharmacological and genetic inhibition of eNOS induced a state of pseudohypoxia, revealed by the stabilization of hypoxia-inducible factor-1 alpha (HIF-1 alpha). Furthermore, NO loss induced a significant decrease in mitochondrial mass and energy production accompanied by a lower O-2 consumption. Notably, very low doses of chronically administered DETA/NO reverted the HIF-1 alpha accumulation, the increased VEGF expression and the stimulated migratory behaviour detected in NO deficient cells.Conclusion: Based on our results, we propose that basal release of NO may act as a negative controller of HIF-1 alpha levels with important consequences for endothelial cell physiology. Moreover, we suggest that our experimental model where eNOS activity was impaired by pharmacological and genetic inhibition may represent a good in vitro system to study endothelial dysfunction.