(−)-Epicatechin induced reversal of endothelial cell aging and improved vascular function: underlying mechanisms

Abstract

The consumption of cocoa products rich in (−)-epicatechin is associated with reduced cardiovascular risk and improved vascular function. However, little is known about (−)-epicatechin's effects on aged endothelium. In order to characterize the health restoring effects of (−)-epicatechin on aged endothelium and identify the underlying mechanisms, we utilized high passage number (i.e. aged) bovine coronary artery endothelial cells and aortas of 3 and 18 month old rats. We evaluated cell senescence (β-galactosidase), nitric oxide (NO) production through the endothelial nitric oxide synthase pathway, mitochondria related endpoints, citrate synthase activity and vascular relaxation. Cells were treated with water or (−)-epicatechin (1 μM) for 48 h and rats orally with either water or (−)-epicatechin (1 mg kg⁻¹ day⁻¹) for 15 days. Senescence associated β-galactosidase levels doubled in aged cells while those treated with (−)-epicatechin only evidenced an ∼40% increase. NO levels in cells decreased by ∼33% with aging and (−)-epicatechin normalized them. Endothelial nitric oxide synthase phosphorylation levels paralleled these results. Aging increased total protein and synthase acetylation levels and (−)-epicatechin partially restored them to those of young cells by stimulating sirtuin-1 binding to the synthase. Phosphorylated sirtuin-1, mitofilin, oxidative phosphorylation complexes and transcriptional factor for mitochondria were reduced by ∼40% with aging and were restored by (−)-epicatechin. (−)-Epicatechin enhanced acetylcholine induced aged aorta vasodilation and stimulated NO levels while reducing blood pressure. In conclusion, (−)-epicatechin reverses endothelial cell aging and restores key control elements of vascular function. These actions may partly explain the epidemiological evidence for the beneficial effects of cocoa consumption on the incidence of cardiac and vascular diseases.