title: Spironolactone inhibits NADPH oxidase-induced oxidative stress and enhances eNOS in human endothelial cells.
abstract: Accumulating evidence indicates that aldosterone plays a critical role in the mediation of oxidative stress and vascular damage. NADPH oxidase has been recognized as a major source of oxidative stress in vasculature. However, the relation between NADPH oxidase in aldosterone-mediated oxidative stress in endothelial cells remains to be ascertained. The present study aimed to investigate the relevant role of NADPH oxidase in aldosterone induced oxidative stress and the functional consequence of this effect on endothelial function. Additionally, we attempted to examine the potential role of the mineralocorticoid receptor (MR) antagonist; spironolactone (spiro) in this scenario. Human umbilical artery endothelial cells (HUAECs) were incubated with aldosterone (100 nmol/L, 24 h) in the absence and presence of Spiro (1 µmol/L). The results showed that aldosterone significantly increased the protein expression of NADPH oxidase subunits (Nox2, p47(phox) and p22(phox)) and that spiro markedly inhibited these changes. Functionally, this was associated with an elevation in NADPH oxidase activity and 3-nitrotyrosine (3-NT) as a biochemical marker of oxidative stress. However, pre-incubation with spiro inhibited these consequences. Moreover, MR protein expression was upregulated by aldosterone whereas this effect was suppressed by Spiro. While aldosterone effectively inhibited endothelial nitric oxide (eNOS) protein expression, pretreatment with spiro markedly restored it to its normal level. In conclusion, the results achieved suggest that aldosterone may play a critical role in NADPH oxidase-mediated oxidative stress resulting in reduced eNOS expression in human endothelial cells. Spiro effectively reversed these consequences, suggesting its potential vasculoprotective effect in endothelial dysfunction.
authors.affiliation: Department of Pharmacology and Toxicology, Faculty of Pharmacy, Minia University, Egypt.
authors.forename: Ashraf
authors.initials: A
authors.lastname: Taye
authors.forename: Henning
authors.initials: H
authors.lastname: Morawietz
keywords: Aldosterone; Endothelium; NADPH oxidase; Oxidative stress; Spironolactone