Skip to main content

Advertisement

SpringerLink
Log in

L-Carnitine ameliorates methotrexate-induced oxidative organ injury and inhibits leukocyte death

  • Published:
Cell Biology and Toxicology Aims and scope Submit manuscript

Abstract

Methotrexate (MTX), a folic acid antagonist widely used for the treatment of a variety of tumors and inflammatory diseases, affects normal tissues that have a high rate of proliferation, including the hematopoietic cells of the bone marrow and the gastrointestinal mucosal cells. To elucidate the role of free radicals and leukocytes in MTX-induced oxidative organ damage and the putative protective effect of L-carnitine (L-Car), Wistar albino rats were administered a single dose of MTX (20 mg/kg) followed by either saline or L-Car (500 mg/kg) for 5 days. After decapitation of the rats, trunk blood was obtained, and the ileum, liver, and kidney were removed for histological examination and for the measurement of malondialdehyde (MDA) and glutathione (GSH) levels, myeloperoxidase (MPO) activity, and collagen content. Our results showed that MTX administration increased the MDA and MPO activities and collagen content and decreased GSH levels in all tissues, while these alterations were reversed in L-Car-treated group. The elevated serum TNF-α level observed following MTX treatment was depressed with L-Car. The oxidative burst of neutrophils stimulated by Annexin V was reduced in the saline-treated MTX group, while L-Car abolished this inhibition. Similarly, flow cytometric measurements revealed that leukocyte apoptosis was increased in MTX-treated animals, while L-Car reversed these effects. Severe degeneration of the intestinal mucosa, liver parenchyma, and glomerular and tubular epithelium observed in the saline-treated MTX group was improved by L-Car treatment. These results suggest that L-Car, possibly via its free radical scavenging and antioxidant properties, ameliorates MTX-induced oxidative organ injury and inhibits leukocyte apoptosis. Thus, supplementation with L-Carnitine as an adjuvant therapy may be promising in alleviating the systemic side-effects of chemotherapeutics.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

ELS:

erythrocyte-lysing solution

GSH:

glutathione (reduced)

GSSG:

glutathione (oxidized)

L-Car:

L-carnitine

MDA:

malondialdehyde

MPO:

myeloperoxidase

MTX:

methotrexate

NAD(P)(H):

nicotine–adenine dinucleotide (phosphate) (reduced form)

ROS:

reactive oxygen species

TNF-α:

tumor necrosis factor α

References

  • Alesci S, De Martino MU, Mirani M, et al. L-Carnitine: a nutritional modulator of glucocorticoid receptor functions. FASEB J. 2003;17:1553–5.

    CAS  PubMed  Google Scholar 

  • Altman GG. Changes in the mucosa of the small intestine following methotrexate administration or abdominal x-irradiation. Am J Anat. 1974;140:263–79.

    Google Scholar 

  • Andrieu-Abadie N, Jaffrezou JP, Hatem S, Laurent G, Levade T, Mercadier JJ. L-Carnitine prevents doxorubicin-induced apoptosis of cardiac myocytes: role of inhibition of ceramide generation. FASEB J. 1999;13:1501–10.

    CAS  PubMed  Google Scholar 

  • Babiak RM, Campello AP, Carnieri EG, Olivera MB. Methotrexate: pentose cycle and oxidative stress. Cell Biochem Funct. 1998;16:283–93.

    Article  CAS  PubMed  Google Scholar 

  • Bertelli A, Conte A, Ronca G. L-Propionyl carnitine protects erythrocytes and low density lipoproteins against peroxidation. Drugs Exp Clin Res. 1994;20:191–7.

    CAS  PubMed  Google Scholar 

  • Beuge JA, Aust SD. Microsomal lipid peroxidation. Methods Enzymol. 1978;52:302–11.

    Google Scholar 

  • Beutler E. Glutathione in red blood cell metabolism. In: A manual of biochemical methods. New York: Grune and Stratton; 1975: 112–14.

    Google Scholar 

  • Bykov I, Jarvelainen H, Lindros K. L-Carnitine alleviates alcohol-induced liver damage in rats: role of tumour necrosis factor-alpha. Alcohol Alcohol. 2003;38:400–6.

    CAS  PubMed  Google Scholar 

  • Caetano NN, Campello AP, Carnieri EGS, Kluppel MLW, Oliveira MBM. Effects of methotrexate (MTX) on NAD(P)+ dehydrogenases of HeLa cells: malic enzymes, 2-oxoluterate and isocitrate dehydrogenases. Cell Biochem Funct. 1997;15:259–64.

    Article  CAS  PubMed  Google Scholar 

  • Calabrese V, Scapagnini G, Ravagna A, et al. Disruption of thiol homeostasis and nitrosative stress in the cerebrospinal fluid of patients with active multiple sclerosis: evidence for a protective role of acetylcarnitine. Neurochem Res. 2003;28:1321–8.

    Article  CAS  PubMed  Google Scholar 

  • Chladek J, Martinkova J, Sispera L. An in vitro study on methotrexate hydroxylation in rat and human liver. Physiol Res. 1997;46:371–9.

    CAS  PubMed  Google Scholar 

  • Demling R, Lalonde C, Knox J, Youn Y, Zhu D, Daryani R. Fluid resuscitation with deferoxamine prevents systemic burn-induced oxidant injury. J Trauma. 1991;31:38–44.

    Google Scholar 

  • Di Giacomo C, Latteri F, Fichera C, et al. Effect of acetyl-L-carnitine on lipid peroxidation and xanthine oxidase activity in rat skeletal muscle. Neurochem Res. 1993;18:1157–62.

    CAS  PubMed  Google Scholar 

  • Eksioglu-Demiralp E, Direskeneli H, Kibaroğlu A, Yavuz Ş, Ergun T, Akoğlu T. Neutrophil activation in Behcet's disease. Clin Exp Rheumatol. 2001;19:S19–24.

    CAS  PubMed  Google Scholar 

  • Elliott GR, Lauwen AP, Bonta IL. The effect of acute feeding of carnitine, acetyl carnitine and propionyl carnitine on basal and A23187-stimulated eicosanoid release from rat carrageenan-elicited peritoneal macrophages. Br J Nutr. 1990;64:497–503.

    CAS  PubMed  Google Scholar 

  • Famularo G, De Simone C, Trinchieri V, Mosca L. Carnitines and its congeners: a metabolic pathway to the regulation of immune response and inflammation. Ann NY Acad Sci. 2004;1033:132–8.

    CAS  PubMed  Google Scholar 

  • Ferrari R, Merli E, Cicchitelli G, Mele D, Fucili A, Ceconi C. Therapeutic effects of L-carnitine and propionyl-L-carnitine on cardiovascular diseases: a review. Ann NY Acad Sci. 2004;1033:79–91.

    CAS  PubMed  Google Scholar 

  • Gao F, Horie T. A synthetic analog of prostaglandin E(1) prevents the production of reactive oxygen species in the intestinal mucosa of methotrexate-treated rats. Life Sci. 2002;71:1091–9.

    Article  CAS  PubMed  Google Scholar 

  • Goldman D. Analysis of the cytotoxic deteminants for methotrexate (NSC-740): a role for free intracellular drug. Cancer Chemother Rep. 1975;6:51–61.

    CAS  Google Scholar 

  • Goto E, Tomojiri S, Okamoto I, Tanaka K. Methotrexate poisoning with acute hepatorenal dysfunction. J Toxicol Clin Toxicol. 2001;39:101–4.

    Article  CAS  PubMed  Google Scholar 

  • Hall PM, Jenner MA, Ahern MJ. Hepatotoxicity in a rat model caused by orally administered methotrexate. Hepatology. 1991;14:906–10.

    CAS  PubMed  Google Scholar 

  • Hersh EM, Wong VG, Henderson ES, Freirich EJ. Hepatotoxic effects of methotrexate. Cancer. 1966;19:600–6.

    CAS  PubMed  Google Scholar 

  • Hillegas LM, Griswold DE, Brickson B, Albrightson-Winslow C. Assesment of myeloperoxidase activity in whole rat kidney. J Pharmacol Methods. 1990;24:285–95.

    Google Scholar 

  • Izgut-Uysal VN, Agac A, Derin N. Effect of L-carnitine on carragenan-induced inflammation in aged rats. Gerontology. 2003;49:287–92.

    PubMed  Google Scholar 

  • Jolivet J, Cowan KH, Curt GA, Clendeninn NJ, Chabner BA. The pharmacology and clinical use of methotrexate. N Engl J Med. 1983;309:1094–104.

    CAS  PubMed  Google Scholar 

  • Kelly GS. L-carnitine: therapeutic applications of a conditionally-essential amino acid. Altern Med Rev. 1998;3:345–60.

    CAS  PubMed  Google Scholar 

  • Kintzel PE. Anticancer drug-induced kidney disorders. Drug Saf. 2001;24:19–38.

    Article  CAS  PubMed  Google Scholar 

  • Kumaran S, Deepak B, Naveen B, Panneerselvam C. Effects of levocarnitine on mitochondrial antioxidant systems and oxidative stress in aged rats. Drugs R D. 2003;4:141–7.

    CAS  PubMed  Google Scholar 

  • Lin HI, Chu SJ, Wang D, Feng NH. Pharmacological modulation of TNF production in macrophages. J Microbiol Immunol Infect. 2004;37:8–15.

    Google Scholar 

  • Longoni B, Giovannini L, Migliori M, Bertelli AA, Bertelli A. Cyclosporine-induced lipid peroxidation and propionyl carnitine protective effect. Int J Tissue React. 1999;21:7–11.

    CAS  PubMed  Google Scholar 

  • Lopez De Leon A, Rojkind M. A simple micromethod for collagen and total protein determination in formalin-fixed parraffin-embedded sections. J Histochem Cytochem. 1985;33:737–43.

    CAS  PubMed  Google Scholar 

  • Maccari F, Arseni A, Chiodi P, Ramacci MT, Angelucci L. Levels of carnitines in brain and other tissues of rats of different ages: effect of acetyl-L-carnitine administration. Exp Gerontol. 1990;25:127–34.

    Article  CAS  PubMed  Google Scholar 

  • Manoli I, De Martino MU, Kino T, Alesci S. Modulatory effects of L-carnitine on glucocorticoid receptor activity. Ann NY Acad Sci. 2004;1033:147–57.

    CAS  PubMed  Google Scholar 

  • Miyazono Y, Gao F, Horie T. Oxidative stress contributes to methotrexate-induced small intestinal toxicity in rats. Scand J Gastroenterol. 2004; 39:1119–27.

    CAS  PubMed  Google Scholar 

  • Nagakubo J, Tomimatsu T, Kitajima M, Takayama H, Aimi N, Horie T. Characteristics of transport of fluoresceinated methotrexate in rat small intestine. Life Sci. 2001;69:739–47.

    Article  CAS  PubMed  Google Scholar 

  • Neuman MG, Cameron RG, Haber JA, Katz GG, Malkiewicz IM, Shear NH. Inducers of cytochrome P450 2E1 enhance methotrexate-induced hepatotoxicity. Clin Biochem. 1999;32:519–36.

    CAS  PubMed  Google Scholar 

  • Paradies G, Petrosillo G, Gadaleta MN, Ruggiero FM. The effect of aging and acetyl-L-carnitine on the pyruvate transport and oxidation in rat heart mitochondria. FEBS Lett. 1999;454:207–9.

    Article  CAS  PubMed  Google Scholar 

  • Pertosa G, Grandaliano G, Simone S, Soccio M, Schena FP. Inflammation and carnitine in hemodialysis patients. J Ren Nutr. 2005;15:8–12.

    Article  PubMed  Google Scholar 

  • Przybtkowski E, Averill-Bates DA. Correlation between glutathione and stimulation of the pentose phosphate cycle in situ in Chinese hamster ovary cells exposed to hydrogen peroxide. Arch Biochem Biophys. 1996;325:91–8.

    Google Scholar 

  • Rani PJ, Panneerselvam C. Effect of L-carnitine on brain lipid peroxidation and antioxidant enzymes in old rats. J Gerontol A Biol Sci Med Sci. 2002;57:B134–7.

    PubMed  Google Scholar 

  • Rauchova H, Koudelova J, Drahota Z, Mourek J. Hypoxia-induced lipid peroxidation in rat brain and protective effect of carnitine and phosphocreatine. Neurochem Res. 2002;27:899–904.

    CAS  PubMed  Google Scholar 

  • Reiter RJ, Tan DX, Sainz RM, Mayo JC, Lopez-Burillo S. Melatonin: reducing the toxicity and increasing the efficacy of drugs. J Pharm Pharmacol. 2002;54:1299–321.

    Article  CAS  PubMed  Google Scholar 

  • Robinson JP. Handbook of flow cytometry methods. Oxidative burst. New York: Wiley-Liss; 1993:145–54.

  • Roenigk HH, Bergfield WF, St. Jacquez R, Owens FJ, Hawk WA. Hepatotoxicity of methotrexate in the treatment of psoriasis. Arch Dermatol. 1971;103:250–61.

    PubMed  Google Scholar 

  • Ross D. Glutathione, free radicals and chemotherapeutic agents. Pharmacol Ther. 1988;37:231–49.

    Article  CAS  PubMed  Google Scholar 

  • Rouse K, Nwokedi E, Woodliff JE, Epstein J, Klimberg VS. Glutamine enhances selectivity of chemotherapy through changes in glutathione metabolism. Ann Surg. 1995;221:420–6.

    CAS  PubMed  Google Scholar 

  • Şener G, Paskaloğlu K, Şatıroglu H, Alican I, Kaçmaz A, Sakarcan A. L-Carnitine ameliorates oxidative damage due to chronic renal failure in rats. J Cardiovasc Pharmacol. 2004;43:698–705.

    PubMed  Google Scholar 

  • Şener G, Toklu H, Kapucu C, et al. Melatonin protects against oxidative organ injury in a rat model of sepsis. Surg Today. 2005;35:52–9.

    PubMed  Google Scholar 

  • Swantek JL, Tsen MF, Cobb MH, Thomas JA. IL-1 receptor-associated kinase modulates host responsiveness to endotoxin. J Immunol. 2000;164:4301–6.

    CAS  PubMed  Google Scholar 

  • Thomas S, Fischer FP, Mettang T, Pauli-Magnus C, Weber J, Kuhlmann U. Effects of L-carnitine on leukocyte function and viability in hemodialysis patients: A double-blind randomized trial. Am J Kidney Dis. 1999;34:678–87.

    CAS  PubMed  Google Scholar 

  • Tobias H, Auerbach R. Hepatotoxicity of long term methotrexate therapy for psoriasis. Arch Intern Med. 1973;132:391–6.

    Article  CAS  PubMed  Google Scholar 

  • van den Bongard HJGD, Manhot RAA, Beijnen JH, et al. Successful rescue with leucovorin and thymidine in a patient with high-dose methotrexate induced acute renal failure. Cancer Chemother Pharmacol. 2001;47:537–40.

    Article  CAS  PubMed  Google Scholar 

  • van't Land B, Meijer HP, Frerich J, et al. Transforming growth factor-beta2 protects the small intestine during methotrexate treatment in rats possibly by reducing stem cell cycling. Br J Cancer. 2002;87:113–8.

    Article  PubMed  Google Scholar 

  • Vanella A, Russo A, Acquaviva R, et al. L-PropionyL-carnitine as superoxide scavenger, antioxidant, and DNA cleavage protector. Cell Biol Toxicol. 2000;16:99–104.

    Article  CAS  PubMed  Google Scholar 

  • Vogel WH, Snyder R, Schulman MP. The inhibition of dehydrogenases by folic acid and several of its analogs. Biochim Biophys Res Common. 1963;10:97–101.

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Pharmacology, School of Pharmacy, Istanbul

    G. Şener Ph.D. (Associate Professor)

  2. Department of Hematology and Immunology, Marmara University, Istanbul

    E. Ekşioğlu-Demiralp & M. Çetiner

  3. Department of Histology-Embryology, Marmara University, Istanbul

    F. Ercan & S. Şirvancı

  4. Department of Physiology, Marmara University, Istanbul

    B. Ç. Yeğen

  5. Division of Biochemistry, Kasimpasa Military Hospital, Istanbul, Trukey

    N. Gedik

  6. Department of Pharmacology, Marmara University, School of Pharmacy, Tıbbiye Cad. 34668, Istanbul, Trukey

    G. Şener Ph.D. (Associate Professor)

Authors
  1. G. Şener Ph.D.
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. E. Ekşioğlu-Demiralp
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. M. Çetiner
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. F. Ercan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. S. Şirvancı
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. N. Gedik
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. B. Ç. Yeğen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to G. Şener Ph.D..

Rights and permissions

Reprints and permissions

About this article

Cite this article

Şener, G., Ekşioğlu-Demiralp, E., Çetiner, M. et al. L-Carnitine ameliorates methotrexate-induced oxidative organ injury and inhibits leukocyte death. Cell Biol Toxicol 22, 47–60 (2006). https://doi.org/10.1007/s10565-006-0025-0

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10565-006-0025-0

Keywords

Search

Navigation