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Involvement of nitric oxide on the pathogenesis of irinotecan-induced intestinal mucositis: role of cytokines on inducible nitric oxide synthase activation

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Abstract

Purpose

Intestinal mucositis and the closely associated diarrhea are common costly side effects of irinotecan. Cytokine modulators, such as thalidomide and pentoxifylline, are found capable of attenuating intestinal mucositis progression. Nitric oxide (NO) seems to be a key mediator of the antineoplastic drug toxicity. The aim of this study was to investigate the role of NO on the pathogenesis of intestinal mucositis, as well as the participation of cytokines upon inducible nitric oxide synthase (iNOS) expression in irinotecan-induced intestinal mucositis.

Methods

iNOS-knockout (iNOS−/−) and C57BL/6 (WT, wild type) animals (n = 5–6) were given either saline or irinotecan (60 mg/kg i.p for 4 days), with or without pretreatment with aminoguanidine (50 mg/kg s.c.), thalidomide (60 mg/kg s.c), infliximab (5 mg/kg i.v.), or pentoxifylline (1.7 mg/kg s.c). On day 5, diarrhea was assessed, and following euthanasia, proximal intestinal samples were obtained for myeloperoxidase (MPO) and iNOS activity, morphometric analysis, western blot and immunohistochemistry to iNOS, cytokine dosage, and for in vitro evaluation of gut contractility.

Results

Irinotecan induced severe diarrhea and intestinal smooth muscle over-contractility, accompanied with histopathological changes. Additionally, increased MPO and iNOS activity and iNOS immunoexpression were found in WT animals treated with irinotecan. The rise in MPO, smooth muscle over-contractility, and diarrhea were abrogated in aminoguanidine-treated and iNOS−/− mice. Moreover, through western blot, we verified that infliximab and pentoxifylline significantly inhibited irinotecan-induced iNOS expression. In addition, cytokine concentration was found only partially decreased in irinotecan-treated iNOS−/− mice when compared with wild-type animals that were given irinotecan.

Conclusions

This study suggests a role of nitric oxide in the pathogenesis of irinotecan-induced intestinal mucositis and also provides evidence for the participation of cytokines on iNOS induction.

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Acknowledgments

We are grateful to José Ivan Rodrigues, Maria Silvandira Freire, and Antônio Ricardo Barreto for technical assistance. This work was supported by CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico), CAPES (Fundação Coordenação de Aperfeiçoamento de Pessoal de Nível Superior), and FUNCAP (Fundação Cearense de Apoio ao Desenvolvimento Científico). G.A.C.B., M.H.L.P., and R.A.R. are recipients of Conselho Nacional de Desenvolvimento Científico e Tecnológico fellowships.

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The authors indicated no potential conflict of interest.

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Authors and Affiliations

  1. Department of Physiology and Pharmacology, Faculty of Medicine, Federal University of Ceará, Rua Cel Nunes de Melo, 1315, Rodolfo Teófilo, Fortaleza, Ceará, 60430-270, Brazil

    Roberto César P. Lima-Júnior, Aline A. Figueiredo, Helano C. Freitas, Maria Luisa P. Melo, Deysi Viviana T. Wong, Caio Abner V. G. Leite, Raul P. Medeiros, Raphael D. Marques-Neto, Mariana L. Vale, Marcellus H. L. P. Souza & Ronaldo A. Ribeiro

  2. Department of Morphology, Faculty of Medicine, Federal University of Ceará, Ceará, Brazil

    Gerly Anne C. Brito & Reinaldo B. Oriá

  3. Department of Pharmacology, School of Medicine of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo, Brazil

    Fernando Q. Cunha

  4. Department of Clinical Oncology, Cancer Institute of Ceará, Fortaleza, Ceará, Brazil

    Ronaldo A. Ribeiro

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  1. Roberto César P. Lima-Júnior
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  2. Aline A. Figueiredo
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  4. Maria Luisa P. Melo
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Correspondence to Ronaldo A. Ribeiro.

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Lima-Júnior, R.C.P., Figueiredo, A.A., Freitas, H.C. et al. Involvement of nitric oxide on the pathogenesis of irinotecan-induced intestinal mucositis: role of cytokines on inducible nitric oxide synthase activation. Cancer Chemother Pharmacol 69, 931–942 (2012). https://doi.org/10.1007/s00280-011-1780-z

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