Acta Pharmacologica Sinica 2008 February; 29 (2): 185-192; doi: 10.1111/j.1745-7254.2008.00749.x

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',7'-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 N^ω-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.

Keywords: aging; cyclooxygenase; endothelium; endothelium-dependent contraction; endothelium-derived contracting factor; oxidative stress; reactive oxygen species; thromboxane-prostanoid receptors