Exposure to diesel exhaust up-regulates iNOS expression in ApoE knockout mice

Traffic related particulate matter air pollution is a risk factor for cardiovascular events; however, the biological mechanisms are unclear. We hypothesize that diesel exhaust (DE) inhalation induces up-regulation of inducible nitric oxide synthase (iNOS), which is known to contribute to vascular dysfunction, progression of atherosclerosis and ultimately cardiovascular morbidity and mortality. Methods: ApoE knockout mice (30-week) were exposed to DE (at 200 mu g/m(3) of particulate matter) or filtered-air (control) for 7 weeks (6 h/day, 5 days/week). iNOS expression in the blood vessels and heart was evaluated by immunohistochemistry and western blotting analysis. To examine iNOS activity, thoracic aortae were mounted in a wire myograph. and vasoconstriction stimulated by phenylephrine (PE) was measured with and without the presence of the specific inhibitor for iNOS (1400W). NF-kappa B (p65) activity was examined by ELISA. The mRNA expression of iNOS and NF-kappa B (p65) was determined by real-time PCR. Results: DE exposure significantly enhanced iNOS expression in the thoracic aorta (4-fold) and heart (1.5 fold). DE exposure significantly attenuated PE-stimulated vasoconstriction by similar to 20%, which was partly reversed by 1400W. The mRNA expression of iNOS and NF-kappa B was significantly augmented after DE exposure. NF-kappa B activity was enhanced 2-fold after DE inhalation, and the augmented NF-kappa B activity was positively correlated with iNOS expression (R(2)=0.5998). Conclusions: We show that exposure to DE increases iNOS expression and activity possibly via NF-kappa B-mediated pathway. We suspect that DE exposure-caused up-regulation of iNOS contributes to vascular dysfunction and atherogenesis, which could ultimately lead to urban air pollution-associated cardiovascular morbidity and mortality. (C) 2011 Elsevier Inc. All rights reserved.