Cancer Chemotherapy and Pharmacology

, Volume 65, Issue 6, pp 1117–1123 | Cite as

Oxidative stress and enhanced paracellular permeability in the small intestine of methotrexate-treated rats

  • Tomoko Maeda
  • Yuko Miyazono
  • Kousei Ito
  • Kazuma Hamada
  • Shuichi Sekine
  • Toshiharu Horie
Original Article
First Online:
Received:
Accepted:

Abstract

Purpose

We previously demonstrated the increase of reactive oxygen species (ROS) production and myeloperoxidase (MPO) activity in the small intestine of methotrexate (MTX)-treated rats. In the present study, we investigated the role of ROS modulating intestinal mucosal permeability in this damage.

Method

MTX (20 mg/kg body weight) was administered to rats intravenously. N-Acetylcysteine (NAC; 80 mg/kg body wt), an antioxidant and a precursor of glutathione (GSH) was administered to rats intraperitoneally to investigate the contribution of ROS to the intestinal permeability enhancement. Intestinal permeability was evaluated by determining that of a poorly absorbable marker, fluorescein isothiocyanate-labeled dextran (FD-4; average molecular mass, 4.4 kDa) using the in vitro everted intestine technique. The occurrence of oxidative stress in the small intestine was assayed by measuring chemiluminescence and thiobarbituric acid reactive substances (TBARS) productions in mucosal homogenates of the small intestine.

Results

The mucosal permeability of FD-4 significantly (p < 0.01) increased in MTX-treated rats compared with control rats, as demonstrated by a twofold increase of FD-4 permeation clearance. This suggests an increase in paracellular permeability. Interestingly, the ROS production was observed preceding the increase of paracellular permeability. Treatment with NAC prevented the MTX-induced ROS production and the increase of paracellular permeability.

Conclusions

NAC protected the small intestine of rats from MTX-induced change in paracellular permeability, suggesting that ROS played an important role in the enhanced paracellular permeability.

Keywords

Methotrexate Reactive oxygen species Intestinal permeability Chemiluminescence N-Acetylcysteine Glutathione 
This is a preview of subscription content, log in to check access

References

  1. 1.
    Thomas HJ (1987) Searching for magic bullet: early approaches to chemotherapy-antifolate, methotrexate. Cancer Res 47:5528–5536Google Scholar
  2. 2.
    Rosen G, Caparros B, Huvos AG, Kosloff C, Nirenberg A, Cacavio A, Marcove RC, Lane JM, Mehta B, Urban C (1982) Preoperative chemotherapy for osteogenic sarcoma: selection of postoperative adjuvant chemotherapy based on the response of the primary tumor to preoperative therapy. Cancer 49:1221–1230CrossRefPubMedGoogle Scholar
  3. 3.
    Ortiz Z, Shea B, Suarez-Almazor ME, Moher D, Wells GA, Tugwell P (1998) The efficacy of folic acid and folinic acid in reducing methotrexate gastrointestinal toxicity in rheumatoid arthritis. A metaanalysis of randomized controlled trials. J Rheumatol 25:36–43PubMedGoogle Scholar
  4. 4.
    Taminiau JA, Gall DG, Hamilton JR (1980) Response of the rat small-intestine epithelium to methotexate. Gut 21:486–492CrossRefPubMedGoogle Scholar
  5. 5.
    Pinkerton CR, Cameron CH, Sloan JM, Glasgow JF, Gwevava NJ (1980) Jejunal crypt cell abnormalities associated with methotrexate treatment in children with acute lymphoblatic leukaemia. J Clin Pathol 35:1272–1277CrossRefGoogle Scholar
  6. 6.
    Beck PL, Wong JF, Li Y, Swaminathan S, Xavier RJ, Devaney KL, Podolsky DK (2004) Chemotherapy- and radiotherapy-induced intestinal damage is regulated by intestinal trefoil factor. Gastroenterology 126:796–808CrossRefPubMedGoogle Scholar
  7. 7.
    Khan SA, Wingard JR (2001) Infection and mucosal injury in cancer treatment. NCI Monogr 29:31–36Google Scholar
  8. 8.
    Keefe DM, Schubert MM, Elting LS, Sonis ST, Epstein JB, Raber-Durlacher JE, Migliorati CA, McGuire DB, Hutchins RD, Peterson DE (2007) Mucositis Study Section of the Multinational Association of Supportive Care in Cancer and the International Society for Oral Oncology updated clinical practice guidelines for the prevention and treatment of mucositis. Cancer 109:820–831CrossRefPubMedGoogle Scholar
  9. 9.
    Gibson RJ, Keefe DM, Clarke JM, Regester GO, Thompson FM, Goland GJ, Edwards BG, Cummins AG (2002) The effect of keratinocyte growth factor on tumour growth and small intestinal mucositis after chemotherapy in the rat with breast cancer. Cancer Chemother Pharmacol 50:53–58CrossRefPubMedGoogle Scholar
  10. 10.
    Howarth GS, Cool JC, Bourne AJ, Ballard FJ, Read LC (1998) Insulin-like growth factor-I (IGF-I) stimulates regrowth of the damaged intestine in rats, when administered following, but not concurrent with, methotrexate. Growth Factors 15:279–292CrossRefPubMedGoogle Scholar
  11. 11.
    Jahovic N, Sener G, Cevik H, Ersoy Y, Arbak S, Yegen BC (2004) Amelioration of methotrexate-induced enteritis by melatonin in rats. Cell Biochem Funct 22:169–178CrossRefPubMedGoogle Scholar
  12. 12.
    Tsurui K, Kosakai Y, Horie T, Awazu S (1990) Vitamin A protects the small intestine from methotrexate-induced damage in rats. J Pharmacol Exp Ther 253:1278–1284PubMedGoogle Scholar
  13. 13.
    Horie T, Nakamaru M, Masubuchi Y (1998) Docosahexaenoic acid exhibits a potent protection of small intestine from methotrexate-induced damage in mice. Life Sci 62:1333–1338CrossRefPubMedGoogle Scholar
  14. 14.
    Horie T, Matsumoto H, Kasagi M, Sugiyama A, Kikuchi M, Karasawa C, Awazu S, Itakura Y, Fuwa T (1999) Protective effect of aged garlic extract on the small intestinal damage of rats induced by methotrexate administration. Planta Med 65:545–548CrossRefPubMedGoogle Scholar
  15. 15.
    Gao F, Nakamaru M, Masubuchi Y, Horie T (2001) Protective effect of a synthetic analog of prostaglandin E1 on the small intestinal damage induced by the administration of methotexate to rats. J Pharm Sci 90:1040–1048CrossRefPubMedGoogle Scholar
  16. 16.
    Miyazono Y, Gao F, Horie T (2004) Oxidative stress contributes to methotrexate-induced small intestinal toxicity in rats. Scand J Gastroenterol 39:1119–1127CrossRefPubMedGoogle Scholar
  17. 17.
    Gao F, Ueda S, Horie T (2001) Effect of a synthetic analog of prostaglandin E1 on the intestinal mucosa of methotrexate-treated rats. Anticancer Res 21:1913–1917PubMedGoogle Scholar
  18. 18.
    Sonis ST, Elting LS, Keefe D, Peterson DE, Schubert M, Hauer-Jensen M, Bekele BN, Raber-Durlacher J, Donnelly JP, Rubenstein EB (2004) Perspectives on cancer therapy-induced mucosal injury: pathogenesis, measurement, epidemiology, and consequences for patients. Cancer 100:1995–2025CrossRefPubMedGoogle Scholar
  19. 19.
    Quiles JL, Huertas JR, Battino M, Mataix J, Ramirez-Tortosa MC (2002) Antioxidant nutrients and adriamycin toxicity. Toxicology 180:79–95CrossRefPubMedGoogle Scholar
  20. 20.
    Conklin KA (2000) Dietary antioxidants during cancer chemotherapy: impact on chemotherapeutic effectiveness and development of side effects. Nutr Cancer 37:1–18CrossRefPubMedGoogle Scholar
  21. 21.
    Dehne N, Lautermann J, Petrat F, Rauen U, de Groot H (2001) Cisplatin ototoxicity: involvement of iron and enhanced formation of superoxide anion radicals. Toxicol Appl Pharmacol 174:27–34CrossRefPubMedGoogle Scholar
  22. 22.
    Hagiwara SI, Ishii Y, Kitamura S (2000) Aerosolized administration of N-acetylcysteine attenuates lung fibrosis induced by bleomycin in mice. Am J Respir Crit Care Med 162:225–231PubMedGoogle Scholar
  23. 23.
    Droge W (2002) Free radicals in the physiological control of cell function. Physiol Rev 82:47–95PubMedGoogle Scholar
  24. 24.
    Nakamaru M, Masubuchi Y, Narimatsu S, Awazu S, Horie T (1998) Evaluation of damaged small intestine of mouse following methotrexate administration. Cancer Chemother Pharmacol 41:98–102CrossRefPubMedGoogle Scholar
  25. 25.
    Gao F, Horie T (2002) A synthetic analog of prostaglandin E(1) prevents the production of reactive oxygen species in the intestinal mucosa of methotrexate-treated rats. Life Sci 71:1091–1099CrossRefPubMedGoogle Scholar
  26. 26.
    Sheth P, Basuroy S, Li C, Naren AP, Rao RK (2003) Role of phosphatidylinositol 3-kinase in oxidative stress-induced disruption of tight junctions. J Biol Chem 278:49239–49245CrossRefPubMedGoogle Scholar
  27. 27.
    Rao RK, Basuroy S, Rao VU, Karnaky KJ Jr, Gupta A (2002) Tyrosine phosphorylation and dissociation of occludin-ZO-1 and E-cadherin-beta-catenin complexes from the cytoskeleton by oxidative stress. Biochem J 368(Pt 2):471–481CrossRefPubMedGoogle Scholar
  28. 28.
    Edens HA, Levi BP, Jaye DL, Walsh S, Reaves TA, Turner JR, Nusrat A, Parkos CA (2002) Neutrophil transepithelial migration: evidence for sequential, contact-dependent signaling events and enhanced paracellular permeability independent of transjunctional migration. J Immunol 169:476–486PubMedGoogle Scholar
  29. 29.
    Zen K, Parkos CA (2003) Leukocyte-epithelial interactions. Curr Opin Cell Biol 15:557–564CrossRefPubMedGoogle Scholar
  30. 30.
    Doluisio JT, Billups NF, Dittert LW, Sugita ET, Swintosky JV (1969) Drug absorption I: an in situ rat gut technique yielding realistic absorption rates. J Pharm Sci 58:1196–1200CrossRefPubMedGoogle Scholar
  31. 31.
    Buege JA, Aust SD (1978) Microsomal lipid peroxidation. Methods Enzymol 52:302–310CrossRefPubMedGoogle Scholar
  32. 32.
    Hartsok A, Nelson WJ (2008) Adherens and tight junctions: structure, function and connections to the actin cytoskeleton. Biochim Biophys Acta 1778:660–669CrossRefGoogle Scholar
  33. 33.
    Li T, Ito K, Sumi S-I, Fuwa T, Horie T (2005) Antiapoptosis action of aged garlic extract (AGE) protects epithelial cells from methotrexate induced injury. Gut 54:1819–1820CrossRefPubMedGoogle Scholar
  34. 34.
    Li T, Ito K, Sumi S-I, Fuwa T, Horie T (2009) Protective effect of aged garlic extract (AGE) on the apoptosis of intestinal epithelial cells caused by methotrexate. Cancer Chemother Pharmacol 63:873–880CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Tomoko Maeda
    • 1
  • Yuko Miyazono
    • 1
  • Kousei Ito
    • 2
  • Kazuma Hamada
    • 1
  • Shuichi Sekine
    • 1
  • Toshiharu Horie
    • 1
  1. 1.Department of Biopharmaceutics, Graduate School of Pharmaceutical SciencesChiba UniversityChibaJapan
  2. 2.Department of Pharmacy, Faculty of MedicineThe University of Tokyo Hospital, The University of TokyoTokyoJapan

Personalised recommendations