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 NG-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-1α (HIF-1α). Furthermore, NO loss induced a significant decrease in mitochondrial mass and energy production accompanied by a lower O2 consumption. Notably, very low doses of chronically administered DETA/NO reverted the HIF-1α 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-1α 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.
Chronic Deficiency of Nitric Oxide affects Hypoxia Inducible Factor-1a (HIF-1a) Stability and Migration in Human Endothelial Cells / M.G. Cattaneo, E. Cappellini, R. Benfante, M. Ragni, F. Omodeo Salè, E. Nisoli, N. Borgese, L.M. Vicentini. - In: PLOS ONE. - ISSN 1932-6203. - 6:12(2011 Dec).
|Titolo:||Chronic Deficiency of Nitric Oxide affects Hypoxia Inducible Factor-1a (HIF-1a) Stability and Migration in Human Endothelial Cells|
CATTANEO, MARIA GRAZIA (Primo)
CAPPELLINI, ELISA (Secondo)
VICENTINI, LUCIA (Ultimo)
|Settore Scientifico Disciplinare:||Settore BIO/14 - Farmacologia|
|Data di pubblicazione:||dic-2011|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1371/journal.pone.0029680|
|Appare nelle tipologie:||01 - Articolo su periodico|
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