title: Chronic blockade of nitric oxide biosynthesis in rats: effect on leukocyte endothelial interaction and on leukocyte recruitment.
abstract: BACKGROUND
  Previous studies showed that animals chronically treated with NG-nitro-L-arginine methyl ester (L-NAME) have a reduced inflammatory reaction. Now the role of L-NAME treatment (20 mg/Kg/day/14 days) on leukocyte mobilisation was assessed in rats.
  
  
  METHODS
  In vivo leukocyte recruitment evoked by Bothrops jararaca venom (BjV) and nitrite/nitrate (NO2-/NO3-; Griess reaction) were evaluated in the air pouch cavity. Haematological parameters were evaluated in the bone marrow and in the peripheral compartment. Microcirculatory blood flow, number of rolling and adhered leukocytes, vascular reactivity and mast cell activity were studied by intravital microscopy. Blood pressure was measured by the tail-cuff method. L-selectin and beta(2) integrin expressions on peripheral and bone marrow leukocytes were quantified by flow cytometry.
  
  
  RESULTS
  When compared with control rats (D-NAME) L-NAME treated rats had reduced PMN cell infiltrate (50%) and NO2-/NO3- (27%) in the air pouch cavity. Rolling leukocytes were decreased (70%) in L-NAME-treated animals, which was reversed by topical application of NO donor (SIN-1). BjV stimulation increased the number of rolling and adhered leukocytes only in control rats. Systemic blood pressure, microcirculatory blood flow and microvascular reactivity was not altered by the treatment. Only the vessel response to acetylcholine was delayed in treated rats. Peripheral PMN cells were increased by L-NAME treatment (100%), but the number of bone marrow cells was not altered. The treatment reduced L-selectin expression on circulating leukocytes, by either with (16%) or without (26%) stimulation with BjV; PMN cells were more affected (32-37%). Impairment of L-selectin expression was also verified in bone marrow cells under stimulation with BjV.
  
  
  CONCLUSIONS
  Results show that this schedule of L-NAME treatment promotes a decrease on L-selectin expression. This effect may promote the standstill of leukocytes in the blood compartment and may be responsible, at least in part, for the observed deficient leukocyte-endothelium interactions with subsequent impairment of leukocyte migration to the inflammatory site.
authors.affiliation: Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, Av. Prof. Lineu Prestes 580, bl. 13B, SP, CEP: 05508-900, São Paulo, Brazil.
authors.forename: S H P
authors.initials: SH
authors.lastname: Farsky
authors.forename: P
authors.initials: P
authors.lastname: Borelli
authors.forename: R A
authors.initials: RA
authors.lastname: Fock
authors.forename: S Z
authors.initials: SZ
authors.lastname: Proto
authors.forename: J M C
authors.initials: JM
authors.lastname: Ferreira
authors.forename: S B V
authors.initials: SB
authors.lastname: Mello
mesh_terms: D000818:Animals; D004730:Endothelium, Vascular / Q000166:cytology* / Q000502:physiology; D004791:Enzyme Inhibitors / Q000008:administration & dosage* / Q000494:pharmacology; D007249:Inflammation; D019041:L-Selectin / Q000378:metabolism; D008297:Male; D008833:Microcirculation; D019331:NG-Nitroarginine Methyl Ester / Q000008:administration & dosage* / Q000494:pharmacology; D020556:Neutrophil Infiltration; D009504:Neutrophils / Q000276:immunology* / Q000378:metabolism / Q000502:physiology; D009569:Nitric Oxide / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D019001:Nitric Oxide Synthase; D051381:Rats; D017208:Rats, Wistar
chemical_list: D004791:Enzyme Inhibitors; D019041:L-Selectin; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D019331:NG-Nitroarginine Methyl Ester