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In the current study we used in vitro and in vivo models to determine the sites of nitric oxide production in rat aortic tissue following cytokine stimulation. In vitro studies in which intact rat aortic rings were incubated with endotoxin (1 microg/mL) or interferon-gamma (600 U/mL) indicated that the expression of inducible nitric oxide synthase (iNOS) activity was increased as measured by Northern blot analysis or determination of nitrite production. In situ hybridization showed iNOS mRNA in the endothelium and adventitia of the incubated aortic rings but not in the media. Immunohistochemical staining showed a similar localization for iNOS protein in the incubated rings. Additional studies were performed in which bacterial endotoxin (4 mg/kg) was administered to rats, and iNOS expression was assayed using in situ hybridization and immunohistochemistry. Clear increases in iNOS mRNA and protein were found in aortic tissue. Endothelial and adventitial cells were the major source of iNOS, with relatively low amounts of iNOS mRNA present in medial smooth muscle, consistent with in vitro findings. These studies indicate that the aortic adventitia is a potential source of NO, and suggest that the adventitial fibroblast may have an important paracrine role in regulating arterial structure and function.

Adventitia as a source of inducible nitric oxide synthase in the rat aorta nitric oxide production by the aortic adventitia