Advertisement

Cancer Chemotherapy and Pharmacology

, Volume 71, Issue 5, pp 1209–1218 | Cite as

A preclinical study on the protective effect of melatonin against methotrexate-induced small intestinal damage: effect mediated by attenuation of nitrosative stress, protein tyrosine nitration, and PARP activation

  • Viswa Kalyan Kolli
  • Indirani Kanakasabapathy
  • Minnie Faith
  • Hemalatha Ramamoorthy
  • Bina Isaac
  • Kasthuri Natarajan
  • Premila AbrahamEmail author
Original Article

Abstract

Purpose

One of the major toxic side effects of methotrexate (MTX) is enterocolitis. To date, there is no efficient standard treatment for this side effect. Nitrosative stress is reported to play a critical role in MTX-induced mucositis. The purpose of this study is to investigate whether pretreatment with melatonin, an inhibitor of nitro-oxidative stress, prevents MTX-induced mucositis in rats.

Methods

Rats were pretreated with melatonin (20 and 40 mg/kg body weight) i.p. daily 1 h before MTX (7 mg/kg body weight) administration for three consecutive days. After the final dose of MTX, the rats were killed and the small intestines were used for analysis.

Results

The small intestines of MTX-treated rats showed moderate to severe injury. The villi were distorted, blunted, and atrophied and focally absent in various segments of the small intestines. Crypt abscesses were also found, suggesting an inflammatory response. Pretreatment with melatonin had a dose-dependent protective effect on MTX-induced mucositis. Morphology was saved to a moderate extent with 20 mg melatonin pretreatment, and near-normal morphology was achieved with 40 mg melatonin pretreatment. Damage to the villi and crypt abscess was reduced. The villi/crypt ratio was almost restored. Melatonin pretreatment protected the small intestines from MTX-induced damage by attenuating nitrosative stress, protein tyrosine nitration and PARP expression.

Conclusion

Because of its versatility in protecting against nitro-oxidative stress and reducing inflammation, we suggest that melatonin could be beneficial in ameliorating MTX-induced enteritis in humans.

Keywords

Anticancer drug Methotrexate Melatonin Small intestine Nitrosative stress 

Notes

Acknowledgments

The authors would like to thank CSIR, New Delhi, for the financial support. Mr. Viswa Kalyan Kolli is a senior research fellow on the project.

Conflict of interest

None.

References

  1. 1.
    Hardeland R, Coto-Montes A, Poeggeler B (2003) Circadian rhythms, oxidative stress and antioxidative defense mechanisms. Chronobiol Int 20:921–962PubMedCrossRefGoogle Scholar
  2. 2.
    Burger D, Travis S (2011) Conventional medical management of inflammatory bowel disease. Gastroenterology 140:1827–1837PubMedCrossRefGoogle Scholar
  3. 3.
    Saibeni S, Bollani S, Losco A, Michielan A, Sostegni R, Devani M, Lupinacci G, Pirola L, Cucino C, Meucci G, Basilisco G, D’Incà R, Bruno S (2012) The use of methotrexate for treatment of inflammatory bowel disease in clinical practice. Dig Liver Dis 44:123–127PubMedCrossRefGoogle Scholar
  4. 4.
    Muscaritoli M, Grieco G, Capria S, Iori AP, Rossi Fanelli F (2002) Nutritional and metabolic support in patients undergoing bone marrow transplantation. Am J Clin Nutr 75:183–190PubMedGoogle Scholar
  5. 5.
    Keefe DM, Gibson RJ, Hauer-Jensen M (2004) Gastrointestinal mucositis. Semin Oncol Nurs 20:38–47PubMedCrossRefGoogle Scholar
  6. 6.
    Nagakubo J, Tomimatsu T, Kitajima M, Takayama H, Aimi N, Horie T (2001) Characteristics of transport of fluoresceinated methotrexate in rat small intestine. Life Sci 69:739–747PubMedCrossRefGoogle Scholar
  7. 7.
    Keefe DM, Cummins AG, Dale BM, Kotasek D, Robb TA, Sage RE (1997) Effect of high-dose chemotherapy on intestinal permeability in humans. Clin Sci (Lond) 92:385–389Google Scholar
  8. 8.
    Pico JL, Avila-Garavito A, Naccache P (1998) Mucositis: its occurrence, consequences, and treatment in the oncology setting. Oncologist 3:446–451PubMedGoogle Scholar
  9. 9.
    Ballabeni V, Ghizzardi P, Cattaruzza F, Bertoni S, Lagrasta CA, Impicciatore M (2006) The selective inhibition of inducible nitric oxide synthase prevents intestinal ischemia-reperfusion injury in mice. Nitric Oxide 14:212–218PubMedCrossRefGoogle Scholar
  10. 10.
    Dong W, Mei Q, Yu J, Xu J-M, Xiang L, Xu Y (2003) Effect of melatonin on the expression of iNOS and COX-2 in rat models of colitis. World J Gastroenterol 9:1307–1311PubMedGoogle Scholar
  11. 11.
    Hosoi T, Goto H, Arisawa T (2001) Role of nitric oxide synthase inhibitor in experimental colitis induced by 2,4,6-trinitrobenzene sulphonic acid in rats. Clin Exp Pharmacol Physiol 28:9–12PubMedCrossRefGoogle Scholar
  12. 12.
    Bian K, Harari Y, Zhong M, Lai M, Castro G, Weisbrodt N, Murad F (2001) Down-regulation of inducible nitric-oxide synthase (NOS-2) during parasite-induced gut inflammation: a path to identify a selective NOS-2 inhibitor. Mol Pharmacol 59:939–947PubMedGoogle Scholar
  13. 13.
    Kolli VK, Abraham P, Rabi S (2008) Methotrexate-induced nitrosative stress may play a critical role in small intestinal damage in the rat. Arch Toxicol 82:763–770PubMedCrossRefGoogle Scholar
  14. 14.
    Leitão RF, Brito GA, Oriá RB, Braga-Neto MB, Bellaguarda EA, Silva JV, Gomes AS, Lima-Júnior RC, Siqueira FJ, Freire RS, Vale ML, Ribeiro RA (2011) Role of inducible nitric oxide synthase pathway on methotrexate-induced intestinal mucositis in rodents. BMC Gastroenterol 11:90–111PubMedCrossRefGoogle Scholar
  15. 15.
    El-Boghdady NA (2011) Protective effect of ellagic acid and pumpkin seed oil against methotrexate-induced small intestine damage in rats. Indian J Biochem Biophys 48:380–387PubMedGoogle Scholar
  16. 16.
    Sewerynek E, Reiter RJ, Melchiorri D (1996) Oxidative damage in the liver induced by ischemia reperfusion: protection by melatonin. Hepatogastroenterology 43:898–905PubMedGoogle Scholar
  17. 17.
    Chang HM, Ling EA, Chen CF, Lue H, Wen CY, Shieh JY (2002) Melatonin attenuates the neuronal NADPH-d/NOS expression in the no dose ganglion of acute hypoxic rats. J Pineal Res 32:65–73PubMedCrossRefGoogle Scholar
  18. 18.
    Ersoz N, Guven A, Cayci T, Uysal B, Turk E, Oztas E, Akgul EO, Korkmaz A, Cetiner S (2009) Comparison of the efficacy of melatonin and 1,400 W on renal ischemia/reperfusion injury: a role for inhibiting iNOS. Ren Fail 31:704–710PubMedCrossRefGoogle Scholar
  19. 19.
    Kelly RW, Amato F, Seamark RF (1984) N-Acetyl-5-methoxy kynurenamine, a brain metabolite of melatonin, is a potent inhibitor of prostaglandin biosynthesis. Biochem Biophys Res Commun 121:372–379PubMedCrossRefGoogle Scholar
  20. 20.
    Szabó C (2006) Poly (ADP-ribose) polymerase activation by reactive nitrogen species—relevance for the pathogenesis of inflammation. Nitric Oxide 14:169–179PubMedCrossRefGoogle Scholar
  21. 21.
    Razzouk BI, Rose SR, Hongeng S, Wallace D, Smeltzer MP, Zacher M, Pui CH, Hudson MM (2007) Obesity in survivors of childhood acute lymphoblastic leukemia and lymphoma. J Clin Oncol 25:1183–1189PubMedCrossRefGoogle Scholar
  22. 22.
    Kehoe JE, Harvey LP, Daly JM (1986) Alteration of chemotherapy toxicity using a chemically defined liquid diet in rats. Cancer Res 46:4047–4052PubMedGoogle Scholar
  23. 23.
    Warden RA, Noltorp RS, Lynn Francis J, Dunkley PR, O’Loughlin EV (1997) Vitamin A deficiency exacerbates methotrexate-induced jejunal injury in rats. J Nutr 127:770–776PubMedGoogle Scholar
  24. 24.
    Ucar M, Korkmaz A, Reiter RJ (2007) Melatonin alleviates lung damage induced by the chemical warfare agent nitrogen mustard. Toxicol Lett 173:124–131PubMedCrossRefGoogle Scholar
  25. 25.
    Chiu CJ, McArdle AH, Brown R, Scott HJ, Gurd FN (1970) Intestinal mucosal lesion in low-flow states. I. A morphological, hemodynamic, and metabolic reappraisal. Arch Surg 101:478–483PubMedCrossRefGoogle Scholar
  26. 26.
    Cuzzocrea S, Zingarelli B, Costantino G (1997) Beneficial effects of 3-aminobenzamide, an inhibitor of poly (ADP-ribose) synthetase in a rat model of splanchnic artery occlusion and reperfusion. Br J Pharmacol 121:1065–1074PubMedCrossRefGoogle Scholar
  27. 27.
    Sastry KV, Moudgal RP, Mohan J, Tyagi JS, Rao GS (2002) Spectrophotometric determination of serum nitrite and nitrate by copper–cadmium alloy. Anal Biochem 306:79–82PubMedCrossRefGoogle Scholar
  28. 28.
    Kupper JH, van Gool L, Muller M (1996) Detection of poly (ADP-ribose) polymerase and its reaction product by immunohistochemistry. Histochem J 28:391–395PubMedCrossRefGoogle Scholar
  29. 29.
    Bubenik GA, Pang SF, Cockshut JR (2000) Circadian variation of portal, arterial and venous blood levels of melatonin in pigs and its relationship to food intake and sleep. J Pineal Res 28:9–15PubMedCrossRefGoogle Scholar
  30. 30.
    Stefulj J, Hörtner M, Ghosh M (2001) Gene expression of the key enzymes of melatonin synthesis in extrapineal tissues of the rat. J Pineal Res 30:243–247PubMedCrossRefGoogle Scholar
  31. 31.
    Bubenik GA, Brown GM (1997) Pinealectomy reduces melatonin levels in the serum but not in the gastrointestinal tract of rats. Biol Signals 6:40–44PubMedCrossRefGoogle Scholar
  32. 32.
    Kvetnoy IM, Ingel IE, Kvetnaia TV, Malinovskaya NK, Rapoport SI, Raikhlin NT, Trofimov AV, Yuzhakov VV (2002) Gastrointestinal melatonin: cellular identification and biological role. Neuro Endocrinol Lett 23:121–132PubMedGoogle Scholar
  33. 33.
    Lee PPN, Pang SF (1993) Melatonin and its receptors in the gastrointestinal tract. Biol Signals 2:181–193PubMedCrossRefGoogle Scholar
  34. 34.
    Pandi-Perumal SR, Srinivasan V, Maestroni GJM, Cardinali DP, Poeggeler B, Hardeland R (2006) Melatonin nature is most versatile biological signal? FEBS J 273:2813–2838PubMedCrossRefGoogle Scholar
  35. 35.
    Seabra ML, Bignotto M, Pinto LR Jr, Tufik S (2000) Randomized, double-blind clinical trial, controlled with placebo, of the toxicology of chronic melatonin treatment. J Pineal Res 29:193–200PubMedCrossRefGoogle Scholar
  36. 36.
    Weishaupt JH, Bartels C, Pölking E, Dietrich J, Rohde G, Poeggeler B, Mertens N, Sperling S, Bohn M, Hüther G, Schneider A, Bach A, Sirén AL, Hardeland R, Bähr M, Nave KA, Ehrenreich H (2006) Reduced oxidative damage in ALS by high-dose enteral melatonin treatment. J Pineal Res 41:313–323PubMedCrossRefGoogle Scholar
  37. 37.
    Nordlund JJ, Lerner AB (1977) The effects of oral melatonin on skin color and on the release of pituitary hormones. J Clin Endocrinol Metab 45:768–774PubMedCrossRefGoogle Scholar
  38. 38.
    Lusardi P, Piazza E, Fogari R (2000) Cardiovascular effects of melatonin in hypertensive patients well controlled by nifedipine: a 24-hour study. Br J Clin Pharmacol 49:423–427PubMedCrossRefGoogle Scholar
  39. 39.
    Lewy AJ, Ahmed S, Sack RL (1996) Phase shifting the human circadian clock using melatonin. Behav Brain Res 73:131–134PubMedCrossRefGoogle Scholar
  40. 40.
    Brzezinski A (1997) Melatonin in humans. N Engl J Med 336:186–195PubMedCrossRefGoogle Scholar
  41. 41.
    Bejarano I, Espino J, Barriga C, Reiter RJ, Pariente JA, Rodríguez AB (2011) Pro-oxidant effect of melatonin in tumour leucocytes: relation with its cytotoxic and pro-apoptotic effects. Basic Clin Pharmacol Toxicol 108:14–20PubMedCrossRefGoogle Scholar
  42. 42.
    Kadoma Y, Fujisawa S (2011) Radical-scavenging activity of melatonin, either alone or in combination with vitamin E, ascorbate or 2-mercaptoethanol as co-antioxidants, using the induction period method. In vivo 25:49–53PubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Viswa Kalyan Kolli
    • 1
  • Indirani Kanakasabapathy
    • 2
  • Minnie Faith
    • 1
  • Hemalatha Ramamoorthy
    • 1
  • Bina Isaac
    • 2
  • Kasthuri Natarajan
    • 1
  • Premila Abraham
    • 1
    Email author
  1. 1.Department of BiochemistryChristian Medial College, BagayamVelloreIndia
  2. 2.Department of AnatomyChristian Medial College, BagayamVelloreIndia

Personalised recommendations