title: Role of inducible nitric oxide synthase pathway on methotrexate-induced intestinal mucositis in rodents.
abstract: BACKGROUND
  Methotrexate treatment has been associated to intestinal epithelial damage. Studies have suggested an important role of nitric oxide in such injury. The aim of this study was to investigate the role of nitric oxide (NO), specifically iNOS on the pathogenesis of methotrexate (MTX)-induced intestinal mucositis.
  
  
  METHODS
  Intestinal mucositis was carried out by three subcutaneous MTX injections (2.5 mg/kg) in Wistar rats and in inducible nitric oxide synthase knock-out (iNOS-/-) and wild-type (iNOS+/+) mice. Rats were treated intraperitoneally with the NOS inhibitors aminoguanidine (AG; 10 mg/Kg) or L-NAME (20 mg/Kg), one hour before MTX injection and daily until sacrifice, on the fifth day. The jejunum was harvested to investigate the expression of Ki67, iNOS and nitrotyrosine by immunohistochemistry and cell death by TUNEL. The neutrophil activity by myeloperoxidase (MPO) assay was performed in the three small intestine segments.
  
  
  RESULTS
  AG and L-NAME significantly reduced villus and crypt damages, inflammatory alterations, cell death, MPO activity, and nitrotyrosine immunostaining due to MTX challenge. The treatment with AG, but not L-NAME, prevented the inhibitory effect of MTX on cell proliferation. MTX induced increased expression of iNOS detected by immunohistochemistry. MTX did not cause significant inflammation in the iNOS-/- mice.
  
  
  CONCLUSIONS
  These results suggest an important role of NO, via activation of iNOS, in the pathogenesis of intestinal mucositis.
authors.affiliation: Department of Morphology, Federal University of Ceará, Fortaleza, Brazil.
authors.forename: Renata Fc
authors.initials: RF
authors.lastname: Leitão
authors.forename: Gerly Ac
authors.initials: GA
authors.lastname: Brito
authors.forename: Reinaldo B
authors.initials: RB
authors.lastname: Oriá
authors.forename: Manuel B
authors.initials: MB
authors.lastname: Braga-Neto
authors.forename: Emmanuelle Al
authors.initials: EA
authors.lastname: Bellaguarda
authors.forename: Johann V
authors.initials: JV
authors.lastname: Silva
authors.forename: Antoniella S
authors.initials: AS
authors.lastname: Gomes
authors.forename: Roberto Cp
authors.initials: RC
authors.lastname: Lima-Júnior
authors.forename: Francisco Jws
authors.initials: FJ
authors.lastname: Siqueira
authors.forename: Rosemeyre S
authors.initials: RS
authors.lastname: Freire
authors.forename: Mariana L
authors.initials: ML
authors.lastname: Vale
authors.forename: Ronaldo A
authors.initials: RA
authors.lastname: Ribeiro
mesh_terms: D000818:Animals; D016923:Cell Death / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000008:administration & dosage; D006146:Guanidines / Q000008:administration & dosage*; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling; D007274:Injections, Intraperitoneal; D019394:Ki-67 Antigen / Q000378:metabolism; D008297:Male; D008727:Methotrexate / Q000009:adverse effects*; D051379:Mice; D018345:Mice, Knockout; D052016:Mucositis / Q000139:chemically induced / Q000188:drug therapy / Q000201:enzymology*; D019331:NG-Nitroarginine Methyl Ester / Q000008:administration & dosage*; D009569:Nitric Oxide / Q000502:physiology; D052247:Nitric Oxide Synthase Type II / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009195:Peroxidase / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D006146:Guanidines; D019394:Ki-67 Antigen; D009569:Nitric Oxide; C002744:3-nitrotyrosine; D014443:Tyrosine; D009195:Peroxidase; D052247:Nitric Oxide Synthase Type II; C004479:pimagedine; D019331:NG-Nitroarginine Methyl Ester; D008727:Methotrexate