TY - JOUR
T1 - Role of inducible nitric oxide synthase pathway on methotrexate-induced intestinal mucositis in rodents
AU - Leitão, Renata F C
AU - Brito, Gerly A C
AU - Oriá, Reinaldo B.
AU - Braga-Neto, Manuel B.
AU - Bellaguarda, Emmanuelle A L
AU - Silva, Johann V.
AU - Gomes, Antoniella S.
AU - Lima-Júnior, Roberto C P
AU - Siqueira, Francisco J W S
AU - Freire, Rosemeyre S.
AU - Vale, Mariana L.
AU - Ribeiro, Ronaldo A.
N1 - Funding Information:
The authors thank Maria Silvandira França Pinheiro, Department of Physiology and Pharmacology, and José Ivan Rodrigues de Sousa, Department of Morphology, Faculty of Medicine, Federal University of Ceará, Brazil, for technical assistance. This work was supported by the Brazilian Agency for Scientific and Technological Development (CNPq) and Fundação Cearense de Apoio ao Desenvolvimento Científico e Tecnológico (FUNCAP).
PY - 2011/8/16
Y1 - 2011/8/16
N2 - 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.Conclusion: These results suggest an important role of NO, via activation of iNOS, in the pathogenesis of intestinal mucositis.
AB - 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.Conclusion: These results suggest an important role of NO, via activation of iNOS, in the pathogenesis of intestinal mucositis.
KW - Aminoguanidine
KW - Methotrexate
KW - Nitric oxide
KW - Nitric oxide synthase
KW - Nφ-Nitro-L-arginine methyl ester
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U2 - 10.1186/1471-230X-11-90
DO - 10.1186/1471-230X-11-90
M3 - Article
C2 - 21846355
AN - SCOPUS:80051644417
SN - 1471-230X
VL - 11
JO - BMC Gastroenterology
JF - BMC Gastroenterology
M1 - 90
ER -