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From the Departments of Physiology, Medicine, and Cellular Biology and Anatomy, Center of Excellence in Arthritis and Rheumatology, LSU Medical Center, Shreveport, La.
Correspondence to Dr D. Neil Granger, Department of Physiology and Biophysics, LSU Medical Center, 1501 Kings Hwy, PO Box 33932, Shreveport, LA 71130-3932.
Abstract The objective of this study was to assess the potential contribution of hydrogen peroxide (H2O2) to the leukocyteendothelial cell adhesion and increased microvascular permeability observed in rat mesenteric venules after inhibition of nitric oxide synthesis with NG-nitro-L-arginine methyl ester (L-NAME). Leukocyte adherence and emigration and leakage of fluorescein isothiocyanatelabeled albumin were monitored in postcapillary venules before and after exposure of the tissue to L-NAME. H2O2 production in mesenteric tissue was monitored by using dihydrorhodamine 123 (DHR), the H2O2-sensitive fluorochrome. L-NAME elicited a rapid increase in both the rate of albumin extravasation and oxidation of DHR, which was followed by an increased adherence and emigration of leukocytes in postcapillary venules. Treatment with either catalase or dimethylthiourea attenuated the L-NAMEinduced oxidative stress, albumin leakage, and leukocyteendothelial cell adhesion. Oxidation of DHR was enhanced in animals treated with either 3-amino-1,2,4-triazole (ATZ), an inhibitor of endogenous catalase, or a combination of ATZ and maleic acid diethyl ester, which depletes intracellular glutathione. Animals receiving a CD11/CD18-specific antibody to prevent leukocyte adhesion/emigration exhibited a reduced oxidation of DHR in response to L-NAME. These findings indicate that most of the H2O2 (and secondarily derived oxidants) generated in mesenteric tissue exposed to an inhibitor of nitric oxide production is due to accumulation of activated leukocytes.
Key Words: leukocyteendothelial cell adhesion vascular permeability platelet-leukocyte aggregation mast cell degranulation dihydrorhodamine 123
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