Oxygen radical inhibition of nitric oxide-dependent vascular function in sickle cell disease

Plasma xanthine oxidase (XO) activity was defined as a source of enhanced vascular superoxide (O 12ptminimal amsmath wasysym amsfonts amssymb amsbsy mathrsfs -69pt document equation*₂^{{-}}equation*document) and hydrogen peroxide (H 2 O 2) production in both sickle cell disease (SCD) patients and knockout-transgenic SCD mice. There was a significant increase in the plasma XO activity of SCD patients that was similarly reflected in the SCD mouse model. Western blot and enzymatic analysis of liver tissue from SCD mice revealed decreased XO content. Hematoxylin and eosin staining of liver tissue of knockout-transgenic SCD mice indicated extensive hepatocellular injury that was accompanied by increased plasma content of the liver enzyme alanine aminotransferase. Immunocytochemical and enzymatic analysis of XO in thoracic aorta and liver tissue of SCD mice showed increased vessel wall and decreased liver XO, with XO concentrated on and in vascular luminal cells. Steady-state rates of vascular O 12ptminimal amsmath wasysym amsfonts amssymb amsbsy mathrsfs -69pt document equation*₂^{{-}}equation*document production, as indicated by coelenterazine chemiluminescence, were significantly increased, and nitric oxide (⋅ NO) -dependent vasorelaxation of aortic ring segments was severely impaired in SCD mice, implying oxidative inactivation of ⋅ NO. Pretreatment of aortic vessels with the superoxide dismutase mimetic manganese 5, 10, 15, 20-tetrakis (N -ethylpyridinium-2-yl) porphyrin markedly decreased O 12ptminimal amsmath wasysym amsfonts amssymb amsbsy mathrsfs -69pt document equation*₂^{{-}}equation*document levels and significantly restored acetylcholine-dependent relaxation, whereas catalase had no effect. These data reveal that episodes of intrahepatic hypoxia-reoxygenation associated with SCD can induce the release of XO into the circulation from the liver. This circulating XO can then bind avidly to vessel luminal cells and impair vascular function by creating an oxidative milieu and catalytically consuming ⋅ NO via O 12ptminimal amsmath wasysym amsfonts amssymb amsbsy mathrsfs -69pt document equation*₂^{{-}}equation*document -dependent mechanisms.