Resveratrol attenuates mitochondrial oxidative stress in coronary arterial endothelial cells

Departments of 1 Physiology, 2 Biochemistry, and 3 Cell Biology and Anatomy and Pediatrics, New York Medical College, Valhalla, New York; 4 Department of Geriatric Medicine, Reynolds Oklahoma Center on Aging, University of Oklahoma Health Science Center, Oklahoma City, Oklahoma; 5 Section on Oxidative Stress Tissue Injury, Laboratory of Physiological Studies, National Institute on Alcohol Abuse and Alcoholism, National Institutes of Health, Bethesda, Maryland; and 6 Department of Internal Medicine, Medical Pharmacology and Physiology and Nutritional Sciences, University of Missouri, Columbia, Missouri Submitted April 20, 2009 ; accepted in final form July 24, 2009 The production of hyperglycemia-induced mitochondrial reactive oxygen species (mtROS) is a key event in the development of diabetic complications. Because resveratrol, a naturally occurring polyphenol, has been reported to confer vasoprotection, improving endothelial function and preventing complications of diabetes, we investigated the effect of resveratrol on mtROS production in cultured human coronary arterial endothelial cells (CAECs). The measurement of MitoSox fluorescence showed that resveratrol attenuates both steady-state and high glucose (30 mM)-induced mtROS production in CAECs, an effect that was prevented by the knockdown of the protein deacetylase silent information regulator 2/sirtuin 1 (SIRT1), an intracellular target of resveratrol. An overexpression of SIRT1 mimicked the effects of resveratrol, attenuating mtROS production. Similar results were obtained in CAECs transfected with mitochondria-targeted H 2 O 2 -sensitive HyPer-Mito fluorescent sensor. Amplex red assay showed that resveratrol and SIRT1 overexpression significantly reduced cellular H 2 O 2 levels as well. Resveratrol upregulated MnSOD expression and increased cellular GSH content in a concentration-dependent manner (measured by HPLC coulometric analysis). These effects were attenuated by SIRT1 knockdown and mimicked by SIRT1 overexpression. We propose that resveratrol, via a pathway that involves the activation of SIRT1 and the upregulation of antioxidant defense mechanisms, attenuates mtROS production, suggesting the potential for new treatment approaches targeting endothelial mitochondria in metabolic diseases. vasoprotection; histone deacetylase; sirtuin 1 Address for reprint requests and other correspondence: A. Csiszar, Dept. of Physiology, New York Medical College, Valhalla, NY 10595 (e-mail: anna_csiszar{at}n