Hydrogen peroxide induces leukocyte rolling: modulation by endogenous antioxidant mechanisms including NO

Abstract

In this study, intravital microscopy was used to examine the mechanisms that regulate H2O2-induced leukocyte rolling within rat mesenteric venules in vivo. H2O2 elicited leukocyte rolling within a narrow response window between 10 and 500 microM H2O2. Continuous superfusion with 100 microM H2O2 induced a large but transient increase in the flux of rolling leukocytes, whereas a short 5-min pulse elicited a sustained increase in rolling flux. Both treatments caused increases in leukocyte adhesion. H2O2-induced increases in leukocyte flux and adhesion could be prevented with an anti-P-selectin antibody. Inhibition of endogenous catalase (aminotriazole), glutathione (diethyl maleate), or nitric oxide (NG-nitro-L-arginine methyl ester) shifted the effective concentration of H2O2; continuous superfusion with 10 microM H2O2 now elicited large and sustained increases in leukocyte rolling flux, whereas 100 microM H2O2 elicited less than optimal responses. Dual antioxidant inhibition further reduced the effective H2O2 concentration to 1 microM H2O2. A nitric oxide donor prevented the increased rolling flux induced by 100 microM H2O2. These findings suggest that endogenous antioxidants are important regulators of H2O2-induced, P-selectin-dependent leukocyte rolling in vivo.