Antioxidant Treatment Prevents Serum Deprivation- and TNF-alpha-Induced Endothelial Dysfunction through the Inhibition of NADPH Oxidase 4 and the Restoration of beta-Oxidation

作者：Scioli Maria Giovanna; Bielli Alessandra; Agostinelli Sara; Tarquini Chiara; Arcuri Gaetano; Ferlosio Amedeo; Costanza Gaetana; Doldo Elena; Orlandi Augusto^*

来源：Journal of Vascular Research, 2014, 51(5): 327-337.

DOI：10.1159/000365926

• Antioxidants
• Endothelial dysfunction
• Oxidative stress
• beta-Oxidation
• Reactive oxygen species
• NADPH oxidase 4

Aims: Oxidative stress plays a pivotal role in the impaired endothelial function occurring in vascular diseases. Antioxidant strategies induce a clinical advantage in patients with endothelial dysfunction and atherosclerosis and protect from oxidative damage, but the underlying molecular mechanisms have been poorly evaluated. The aim of this study was to analyze the effects and mechanisms of action of antioxidant regimens on endothelial function. Methods and Results: Antioxidant efficacy of N-acetylcysteine, ascorbic acid and propionyl-L-carnitine was evaluated in serum-deprived and TNF-alpha-stimulated human umbilical vein endothelial cells in vitro. Cell adhesion molecule (CAM) expression was evaluated by blot and real-time PCR, and inflammatory cytokine secretion was evaluated by ELISA; leukocyte adhesion and reactive oxygen species assays and NADPH oxidase 4 isoform (Nox4) expression analyses by blots were also performed. Antioxidant pretreatment restored serum-deprived and TNF-alpha-induced impaired mitochondrial beta-oxidation by reducing flavin adenine dinucleotide level and counteract-ing increased CAM and Nox4 expression, leukocyte adhesion and inflammatory cytokine secretion. Specific inhibition by plumbagin and siNox4 prevented TNF-alpha- and serum deprivation-induced detrimental effects, confirming that endothelial oxidative stress and inflammation were Nox4 dependent. Conclusions: Our findings documented Nox4 as a main actor in oxidative stress-induced endothelial dysfunction and further clarify the molecular basis of antioxidant treatment efficacy.

• 出版日期2014