Yi Shi1, Ricky Y K Man, Paul M Vanhoutte. 1. Department of Pharmacology, The University of Hong Kong, Hong Kong, China. yi.shi@access.uzh.ch
Abstract
AIM: The present experiments were designed to study the changes in endothelium-dependent contractions with aging. METHODS: The rat femoral arteries of 20-week and 1-year-old rats with and without endothelium were suspended in organ chambers to record isometric tension. The production of oxygen-derived free radicals in the endothelium was measured with 2,7o-dichlorodihydrofluorescein diacetate (DCF) using confocal microscopy. Protein presences were determined by Western blotting. RESULTS: In the arteries from the 1-year-old rats, endothelium-dependent relaxations to A23187 were reduced, but the endothelium-dependent contractions to A23187 (in the presence of Nomega-nitro-L-arginine methyl ester hydrochloride [L-NAME; an inhibitor of nitric oxide synthase]) were augmented, demonstrating endothelial dysfunction with aging. Indomethacin normalized the responses, suggesting that a cyclooxygenase (COX)-dependent contraction is prominent in aging. The endothelium-dependent contractions were also prevented by terutroban (a blocker of thromboxane-prostanoid receptors), confirming the activation of thromboxane-prostanoid receptors on vascular smooth muscle. Valeryl salicylate and NS-398 (preferential inhibitors of COX-1 and COX-2, respectively) partially reduced the response, indicating that both COX-1 and COX-2 are involved. Western blotting confirmed the upregulation of both isoforms in the arteries of the 1-year-old rats. In the presence of L-NAME, A23187 increased the DCF fluorescence in the endothelium, demonstrating that the production of oxygen-derived free radicals contributes to endothelium-dependent contractions. The activity of catalase was reduced in the arteries with endothelium of 1-year-old rats, indicating that hydrogen peroxide is the likely mediator of increased oxidative stress in the aging endothelium. CONCLUSION: Endothelium-dependent contractions are augmented with aging. Oxidative stress potentiates the response, and both COX-1 and COX-2 are involved.
AIM: The present experiments were designed to study the changes in endothelium-dependent contractions with aging. METHODS: The rat femoral arteries of 20-week and 1-year-old rats with and without endothelium were suspended in organ chambers to record isometric tension. The production of oxygen-derived free radicals in the endothelium was measured with 2,7o-dichlorodihydrofluorescein diacetate (DCF) using confocal microscopy. Protein presences were determined by Western blotting. RESULTS: In the arteries from the 1-year-old rats, endothelium-dependent relaxations to A23187 were reduced, but the endothelium-dependent contractions to A23187 (in the presence of Nomega-nitro-L-arginine methyl ester hydrochloride [L-NAME; an inhibitor of nitric oxide synthase]) were augmented, demonstrating endothelial dysfunction with aging. Indomethacin normalized the responses, suggesting that a cyclooxygenase (COX)-dependent contraction is prominent in aging. The endothelium-dependent contractions were also prevented by terutroban (a blocker of thromboxane-prostanoid receptors), confirming the activation of thromboxane-prostanoid receptors on vascular smooth muscle. Valeryl salicylate and NS-398 (preferential inhibitors of COX-1 and COX-2, respectively) partially reduced the response, indicating that both COX-1 and COX-2 are involved. Western blotting confirmed the upregulation of both isoforms in the arteries of the 1-year-old rats. In the presence of L-NAME, A23187 increased the DCF fluorescence in the endothelium, demonstrating that the production of oxygen-derived free radicals contributes to endothelium-dependent contractions. The activity of catalase was reduced in the arteries with endothelium of 1-year-old rats, indicating that hydrogen peroxide is the likely mediator of increased oxidative stress in the aging endothelium. CONCLUSION: Endothelium-dependent contractions are augmented with aging. Oxidative stress potentiates the response, and both COX-1 and COX-2 are involved.
Authors: Markus M Bachschmid; Stefan Schildknecht; Reiko Matsui; Rebecca Zee; Dagmar Haeussler; Richard A Cohen; David Pimental; Bernd van der Loo Journal: Ann Med Date: 2012-03-01 Impact factor: 4.709
Authors: Anne D Thuesen; Kenneth Andersen; Kristina S Lyngsø; Mark Burton; Charlotte Brasch-Andersen; Paul M Vanhoutte; Pernille B L Hansen Journal: Pflugers Arch Date: 2017-10-29 Impact factor: 3.657