Resveratrol attenuates mitochondrial ROS generation in endothelial cells treated with cigarette smoke extract

Abstract

Previously we have shown that water soluble components of cigarette smoke (CS) up‐regulate NAD(P)H oxidase‐dependent ROS production in endothelial cells. In the present study we tested the effect of CS on mitochondrial ROS production in cultured coronary arterial endothelial cells (CAECs) and determined whether the sirtuin activator resveratrol (RES) protects CAECs against the deleterious effects of CS. Mitochondrial O2.‐ production was assessed in CS extract treated HCAECs by flow cytometry and confocal microscopy using MitoSOX Red. CS extract (0.04 to 4 ug/mL) elicited dose‐dependent increases in mitochondrial O2.‐ generation which were prevented by resveratrol pre‐treatment (1–10 uM; for 24 h). At higher concentrations CSE also elicited apoptosis in CAECs (as reflected by the increased cellular annexin V staining and caspase 3 activity), which was also prevented by resveratrol. Thus, RES at nutritionally relevant concentrations attenuates CS‐induced mitochondrial oxidative stress and protects endothelial cells from apoptotic cell death.