Pravastatin Counteracts Angiotensin II-Induced Upregulation and Activation of NADPH Oxidase at Plasma Membrane of Human Endothelial Cells

Abstract

Endothelial dysfunction has been linked to reactive oxygen species (ROS) production by nicotinamide adenine dinucleotide phosphate reduced (NADPH) oxidase. Angiotensin II (ANG), which levels are elevated in some cardiovascular diseases, can stimulate this enzyme, whereas statins have been demonstrated pleiotropic effects related with the restoration of endothelial function. Therefore, our purpose was to study the mechanism of the possible beneficial effects of pravastatin on ANG-activated human umbilical vein endothelial cells (HUVEC). ANG induced an increase in the extracellular superoxide anion produced by NADPH oxidase but had no effect in the intracellular ROS production. Pravastatin, which alone did not have any effect on ROS production, totally blocked the stimulating effects of ANG when combined with it. These effects were not due to a direct action of ANG or pravastatin on the activity of NADPH oxidase measured in HUVEC lisates. On the contrary, the results correlated well with other effects of ANG: a Nox4 and p22phox upregulated expression and an enhanced Nox4 translocation to the cell membrane. All these effects were inhibited by pravastatin, which had no effect when incubated alone. These data reveal for the first time that pravastatin interrupts the signaling pathway activated by ANG that leads to an enhanced NADPH oxidase activity at the cell membrane of HUVEC. For that, pravastatin inhibits ANG-induced upregulation of catalytic NADPH oxidase subunits and blocks the migration of them to the endothelial cell membrane.

Attenuation by Statins of Membrane Raft-Redox Signaling in Coronary Arterial Endothelium

“…Simvastatin was activated by opening the lactone ring by dissolving in 95% ethanol and 0.1 N NaOH, heating at 50°C for 2 hours, and neutralizing with HCl to pH 7.2, as described previously (Gerson et al, 1989;Tawfik et al, 2006). The doses of statins were chosen on the basis of previous studies (Dje N'Guessan et al, 2009;Alvarez et al, 2010) and our observations that statins at these doses did not significantly induced morphologic signs of cytotoxicity in HCAECs. Some groups of cells were pretreated with mevalonate (10 mM), farnesol (10 mM), geranylgeraniol (10 mM), methyl-b-cyclodextrin (1 mM), or filipin (1 mg/ml; all reagents were purchased from Sigma-Aldrich) for 1 hour.…”

“…By interfering with cholesterol biosynthesis and lowering plasma membrane cholesterol levels, statins were shown to decrease the expression of caveolin-1, resulting in eNOS activation and nitric oxide production (Mason et al, 2004). Second, by preventing isoprenylation of Rac1, which is important for NADPH oxidase activation, recent studies have revealed that statins inhibit O 2 .2 formation in ECs after stimulation of injury factors or under pathologic conditions, such as angiotensin II, homocysteine, and hyperglycemia (Wagner et al, 2000;Vecchione et al, 2007;Briones et al, 2009;Alvarez et al, 2010;Bao et al, 2010c). However, it remains unknown how statins alter the assembling and aggregation of NADPH oxidase subunits and, thereby, affect its activity to produce O 2 .2 in addition to their effect on Rac1 or eNOS.…”

Pharmacological Actions of Statins: A Critical Appraisal in the Management of Cancer

“…Moreover, pravastatin and fluvastatin had a direct scavenging radical activity Kassan et al, 2010). Pravastatin was also reported to inhibit the stimulatory activity of angiotensin II on NADPH oxidase, thereby contrasting the production of superoxide radicals (Alvarez et al, 2010).…”

Atorvastatin alleviates iodinated contrast media-induced cytotoxicity in human proximal renal tubular epithelial cells

“…A number of retrospective and prospective studies have demonstrated the potential protective effect of statins on CIN (9)(10)(11). In addition, statins have been reported to exhibit antioxidant effects through decreasing the mRNA expression of NOX4 and p22phox (12,13). These findings suggest that statins may exhibit a protective effect in CIN through decreasing the expression of NOX4 and p22phox in kidney cells.…”