We aimed to elucidate the possible role of phenotypic alterations and oxidative stress in age-related endothelial dysfunction of coronary arterioles.

Arterioles were isolated from the hearts of young adult (Y, 14 weeks) and aged (A, 80 weeks) male Sprague-Dawley rats.

For videomicroscopy, pressure-induced tone of Y and A arterioles and their passive diameter did not differ significantly.

In A, arterioles L-NAME (a NO synthase blocker)–sensitive flow-induced dilations were significantly impaired (Y: 41±8% versus A: 3±2%), which could be augmented by superoxide dismutase (SOD) or Tiron (but not l-arginine or the TXA2 receptor antagonist SQ29,548).

For lucigenin chemiluminescence, O2·− generation was significantly greater in A than Y vessels and could be inhibited with SOD and diphenyliodonium.

NADH-driven O2·− generation was also greater in A vessels.

Both endothelial and smooth muscle cells of A vessels produced O2·− (shown with ethidium bromide fluorescence).

For Western blotting, expression of eNOS and COX-1 was decreased in A compared with Y arterioles, whereas expressions of COX-2, Cu/Zn-SOD, Mn-SOD, xanthine oxidase, and the NAD(P)H oxidase subunits p47phox, p67phox, Mox-1, and p22phox did not differ.

Aged arterioles showed an increased expression of iNOS, confined to the endothelium.

Decreased eNOS mRNA and increased iNOS mRNA expression in A vessels was shown by quantitative RT-PCR.

In vivo formation of peroxynitrite was evidenced by Western blotting, and immunohistochemistry showing increased 3-nitrotyrosine content in A vessels.

Thus, aging induces changes in the phenotype of coronary arterioles that could contribute to the development of oxidative stress, which impairs NO-mediated dilations.