IMPACT OF ANTIOXIDANT SUPPLEMENTATION ON TOXICITY OF METHOTREXATE-AN IN VITRO STUDY ON ERYTHROCYTES USING VITAMIN E
Abstract
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Objective: Methotrexate (MTX) is an antimetabolite used widely in cancer and autoimmune diseases as an inhibitor of the enzyme dihydrofolate
reductase. Treatment regimens with MTX lead to the formation of reactive oxygen species and are involved in drug-induced toxicity presenting severe
side effects. In the present investigation, red blood cells were used as a model to understand drug-mediated hemolysis and alterations in antioxidant
defense system stimulated by MTX. This effort attempts to evaluate, whether pre-supplementation of vitamin E can modulate drug induced toxicity.
Methods: Blood samples from healthy subjects were collected and processed to obtain 10% RBC solution in saline. This solution was further treated
with 80μM MTX in presence and absence of 90μM vitamin E respectively. Treated and control RBC solutions were used to assess percent hemolysis,
levels of lipid peroxides, GSH, lactate dehydrogenase and activities of specific antioxidant enzymes.
Results: Our study reveals a significant increase in lipid peroxides, reduced glutathione GSH reductase activity after incubation with MTX, lactate
dehydrogenase, and a considerable decline in catalase, superoxide dismutase, GSH peroxidase, GSH S-transferase, and GSH reductase after incubation
with MTX.
Conclusion: Ameliorative effect of vitamin E supplementation reduces oxidative stress and restores the activities of these antioxidant enzymes,
thereby demonstrating the protection rendered by vitamin E. Our data indicates that vitamin E administration during chemotherapy is effective in
modulating the chemotherapy-induced side effects thus stressing on the importance of vitamin E supplementation in combination with chemotherapy
during cancer treatments.
Keywords: Antioxidant, Lipid peroxides, Superoxide dismutase, Glutathione, Catalase, Red blood cell.
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Allegra CJ, Fine RL, Drake JC, Chabner BA. The effect of methotrexate on intracellular folate pools in human MCF-7 breast cancer cells. Evidence for direct inhibition of purine synthesis. J Biol Chem 1986;261(14):6478-85.
Cogan E. Methotrexate treatment in non-cancerous disorders. Rev Med Brux 1995;16(4):300-3.
Gaies E, Jebabli N, Trabelsi S, Salouage I, Charfi R, Lakhal M, et al. Methotrexate side effects: Review article. J Drug Metab Toxicol 2012;3(4):2-5.
Cronstein BN. Low-dose methotrexate: A mainstay in the treatment of rheumatoid arthritis. Pharmacol Rev 2005;57(2):163-72.
Kremer JM. Toward a better understanding of methotrexate. Arthritis Rheum 2004;50(5):1370-82.
Singh K, Malviya A, Bhori M, Marar T. An in vitro study of the ameliorative role of a-tocopherol on methotrexate-induced oxidative stress in rat heart mitochondria. J Basic Clin Physiol Pharmacol 2012;23(4):163-8.
Coleshowers CL, Oguntibeju OO, Ukpong M, Truter EJ. Effects of methotrexate on antioxidant enzyme status in a rodent model. Med Tech South Afr 2010;24(1):5-9.
Weinblatt ME, Fraser P. Elevated mean corpuscular volume as a predictor of hematologic toxicity due to methotrexate therapy. Arthritis Rheum 1989;32(12):1592-6.
Miyazono Y, Gao F, Horie T. Oxidative stress contributes to methotrexate-induced small intestinal toxicity in rats. Scand J Gastroenterol 2004;39(11):1119-27.
Devrim E, Cetin R, Kiliçoglu B, Ergüder BI, Avci A, Durak I. Methotrexate causes oxidative stress in rat kidney tissues. Ren Fail 2005;27:771-3.
Devasagayam TP, Tilak JC, Boloor KK, Sane KS, Ghaskadbi SS, Lele RD. Free radicals and antioxidants in human health: Current status and future prospects. J Assoc Physicians India 2004;52:794-804.
Brigelius-Flohé R, Kelly FJ, Salonen JT, Neuzil J, Zingg JM, Azzi A. The European perspective on vitamin E: Current knowledge and future research. Am J Clin Nutr 2002;76(4):703-16.
Peralta EA, Viegas ML, Louis S, Engle DL, Dunnington GL. Effect of vitamin E on tamoxifen-treated breast cancer cells. Surgery 2006;140(4):607-14.
Nayma S, Noorzahan B, Shelina B, Sultana F, Ali T. Effects of vitamin E supplementation on some aspects of hematological variables in patients of hemolytic anemia with glucose 6 phosphate dehydrogenase (G6PD) deficiency. Bangladesh J Physiol Pharmacol 2006;22(1,2):12‑7.
Lamson DW, Brignall MS. Antioxidants in cancer therapy; their actions and interactions with oncologic therapies. Altern Med Rev 1999;4(5):304-29.
Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 1979;95(2):351‑8.
Misra HP, Fridovich I. The generation of superoxide radical during the autoxidation of hemoglobin. J Biol Chem 1972;247(21):6960-2.
Claiborne A. Catalase activity. In: Greenwald RA, editor. Handbook of Methods for Oxygen Radical Research. Vol. 1. Boca Raton: CRC Press; 1985. p. 283-4.
Habig WH, Pabst MJ, Jakoby WB. Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem 1974;249(22):7130-9.
Paglia DE, Valentine WN. Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J Lab Clin Med 1967;70(1):158-69.
Carlberg I, Mannervik B. Purification and characterization of glutathione reductase from calf liver. An improved procedure foraffinity chromatography on 2’,5’-ADP-Sepharose 4B. Anal Biochem 1981;116:531-6.
Moron MS, Depierre JW, Mannervik B. Levels of glutathione, glutathione reductase and glutathione S-transferase activities in rat lung and liver. Biochim Biophys Acta 1979;582(1):67-78.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193(1):265-75.
Uzar E, Sahin O, Koyuncuoglu HR, Uz E, Bas O, Kilbas S, et al. The activity of adenosine deaminase and the level of nitric oxide in spinal cord of methotrexate administered rats: Protective effect of caffeic acid phenethyl ester. Toxicology 2006;218(2-3):125-33.
Bhogade RB, Suryakar AN, Joshi NG, Patil RY. Effect of vitamin E supplementation on oxidative stress in hemodialysis patients. Indian J Clin Biochem 2008;23(3):233-7.
Marar T. Amelioration of glucose induced hemolysis of human erythrocytes by vitamin E. Chem Biol Interact 2011;193(2):149-53.
Singh K, Pillai V, Marar T. Influence of vitamin E on camptothecin induced oxidant injury: An in vitro study on erythrocytes. J Pharm Res 2012;5(12):3116-9.
Singh K, Kaur R, Marar T. Ameliorative effect of vitamin E on chemotherapy induced side effects in rat liver. J Pharmacol Toxicol 2011;6(5):01-12.
Niki E, Kawakami A, Saito M, Yamamoto Y, Tsuchiya J, Kamiya Y. Effect of phytyl side chain of vitamin E on its antioxidant activity. J Biol Chem 1985;260:2191-6.
Krishnamurthy S, George T, Jayanthibai N. Effect of retinol on the hemolysis and lipid peroxidation in vitamin Ε deficiency. J Biosci 1984;6(3):325-30.
Sudha K, Rao AV, Rao S, Rao A. Lipid peroxidation, hemolysis and antioxidant enzymes of erythrocytes in stroke. Indian J Physiol Pharmacol 2004;48(2):199-205.
Wang F, Wang T, Lai J, Li M, Zou C. Vitamin E inhibits hemolysis induced by hemin as a membrane stabilizer. Biochem Pharmacol 2006;71(6):799-805.
Cruz CS, Wimberley PD, Johansen K, Friis-Hansen B. The effect of vitamin E on erythrocyte hemolysis and lipid peroxidation in newborn premature infants. Acta Paediatr Scand 1983;72(6):823-6.
Vidyasagar J, Karunakar N, Reddy MS, Rajnarayana K, Surender T, Krishna DR. Oxidative stress and antioxidant status in acute organophosphorous insecticide poisoning. Indian J Pharmacol 2002;36(2):76-9.
Abdel-Ghaffar FR, El-Elaimy IA, El-Dougdoug KA, Nassar HI. Protective and modulatory effects of curcumin and L-carnitine against methotrexate-induced oxidative stress in albino rats. Res J Pharm Biol Chem Sci 2013;4:744-54.
Jacob HS, Lux SE. Degradation of membrane phospholipids and thiols in peroxide hemolysis: Studies in vitamin E deficiency. Blood 1968;32(4):549-68.
Brownlee NR, Huttner JJ, Panganamala RV, Cornwell DG. Role of vitamin E in glutathione-induced oxidant stress: Methemoglobin, lipid peroxidation, and hemolysis. J Lipid Res 1977;18(5):635-44.
Jovanovic P, Zoric L, Stefanovic I, Dzunic B, Djordjevic-Jocic J, Radenkovic M, et al. Lactate dehydrogenase and oxidative stress activity in primary open-angle glaucoma aqueous humour. Bosn J Basic Med Sci 2010;10(1):83-8.
Al-Abrash AS, Al-Quobaili FA, Al-Akhras GN. Catalase evaluation in different human diseases associated with oxidative stress. Saudi Med J 2000;21(9):826-30.
Fridovich I. Superoxide dismutase. Annu Rev Biochem 1975;44:147‑59.
Pigeolet E, Corbisier P, Houbion A, Lambert D, Michiels C, Raes M, et al. Glutathione peroxidase, superoxide dismutase, and catalase inactivation by peroxides and oxygen derived free radicals. Mech Ageing Dev 1990;51(3):283-97.
Little C, O’Brien PJ. An intracellular GSH-peroxidase with a lipid peroxide substrate. Biochem Biophys Res Commun 1968;31(2):145‑50.
Selvakumar E, Prahalathan C, Mythili Y, Varalakshmi P. Mitigation of oxidative stress in cyclophosphamide-challenged hepatic tissue by DL-alpha-lipoic acid. Mol Cell Biochem 2005;272(1-2):179-85.
Halani CG, Dewani AP, Kale MK, Umathe SN, Bhursari KP, Duragkar NJ. Effect of vitamin-E on oxidative stress in diverse thyroid states. Int J Pharm Sci Nanotechnol 2008;1:195-8.
Parimal C, Michael S. Oxidant/anti-oxidant status in rats exposed to simulated weightlessness by hind-limb unloading and reloading. Open Clin Chem J 2008;1:47-56.
Geetha A, Lakshmi Priya MD, Jeyachristy SA, Surendran R. Level of oxidative stress in the red blood cells of patients with liver cirrhosis. Indian J Med Res 2007;126(3):204-10.
Gradinski B, Stojevic Z, Tur SM, Balenovic T, Pirsljin J, Tuk MZ. In vitro susceptibility of duck, chicken, and pig erythrocyte lipids to peroxidation. Vet Med Czech 2002;47:303-8.
Metin G, Atukeren P, Gümüstas MK, Belce A, Kayserilioglu A. The effect of vitamin E treatment on oxidative stress generated in trained rats. Tohoku J Exp Med 2002;198(1):47-53.
Seven A, Guzel S, Seymen O, Civelek S, Bolayirli M, Uncu M, et al. Effects of vitamin E supplementation on oxidative stress in streptozotocin induced diabetic rats: Investigation of liver and plasma. Yonsei Med J 2004;45:703-10.
Hayes JD, Pulford DJ. The glutathione S-transferase supergene family: Regulation of GST and the contribution of the isoenzymes to cancer chemoprotection and drug resistance. Crit Rev Biochem Mol Biol 1995;30(6):445-600.
Gutierrez-Correa J, Stoppani AO. Inactivation of yeast glutathione reductase by Fenton systems: Effect of metal chelators, catecholamines and thiol compounds. Free Radic Res 1997;27(6):543-55.
Muller EG. A glutathione reductase mutant of yeast accumulates high levels of oxidized glutathione and requires thioredoxin for growth. Mol Biol Cell 1996;7(11):1805-13.
Brigelius R. Mixed disulfides: Biological functions and increase in oxidative stress. In: Sies H, editor. Oxidative Stress. London: Academic Press; 1985. p. 243-72.
Chahbouni M, Escames G, Lopez LC, Sevilla B, Doerrier C, Muñoz-Hoyos A, et al. Melatonin treatment counteracts the hyperoxidative status in erythrocytes of patients suffering from Duchenne muscular dystrophy. Clin Biochem 2011;44(10-11):853-8.
Babiak RM, Campello AP, Carnieri EG, Oliveira MB. Methotrexate: Pentose cycle and oxidative stress. Cell Biochem Funct 1998;16(4):283‑93.
Caetano NN, Campello AP, Carnieri EG, Kluppel ML, Oliveira MB. Effect of methotrexate (MTX) on NAD (P) + dehydrogenases of HeLa cells: Malic enzyme, 2-oxoglutarate and isocitrate dehydrogenases. Cell Biochem Funct 1997;15(4):259-64.
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