In vivo nitric oxide production and iNOS expression in X-ray irradiated mouse skin

Inducible nitric oxide synthase (iNOS) and NO have been suggested to be involved in acute radiation response in tissues such as the liver, intestine, colon, and brain. However, direct measurement of NO and iNOS in ionizing radiation-induced skin inflammatory reactions is not reported yet. We show here for the first time, by in vivo experiments, that X-ray irradiated mouse skin generates NO with concomitant expression of iNOS at both the mRNA and protein levels. When irradiated at 50 Gy, iNOS mRNA appeared at day 8 post-irradiation, whereas iNOS protein could be detected only at day 14. No iNOS protein was detectable however for the mice receiving 5 or 15 Gy irradiation, even at day 14. Skin inflammatory reactions were observed at day 8 post-irradiation as an increase in skin thickness, which increased further by day 14. Histological observations showed acute inflammatory responses. The parallel relationship between iNOS induction and the onset of skin inflammatory reactions suggests the involvement of iNOS and NO in the skin damage. Immunohistochemical staining showed the localization of iNOS at skin erosion areas, exudate and infiltrating cells. Taken together, these findings suggest that iNOS induction and NO production in X-irradiated skin are relatively early events in skin inflammatory reactions and are probably secondary rather than primary reactions of irradiation.