Vol 21, No 1 (2018)
Research paper
Published online: 2017-12-13

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The use of 99mTc-phytate for assessment the protective effect of vitamin E against hepatotoxicity induced by methotrexat in rat

Hossein Amirfakhrian, Seyed Mohammad Abedi, Hossein Sadeghi, Soheil Azizi, Seyed Jalal Hosseinimehr
Pubmed: 29319136
Nucl. Med. Rev 2018;21(1):8-13.


In this study, we investigated the protective effect of vitamin E against methotrexate (MTX)-induced hepatotoxicity by quantitative liver 99mTc-phytate uptake and liver imaging and to compare its effect with histopathology in rat. Rats were divided into five groups as control, solvent, Vit E (100 mg/kg), MTX (20 mg/kg), Vit E + MTX and. Vit E was intraperitoneally administrated for 17 days before MTX injection and continued for 4 days. 99mTc-phytate was injected through the tail of rats after the drug administration. The percentage of the injected dose per gram of liver and spleen tissues (%ID/g) was calculated. Liver imaging was obtained with gamma camera. In other experiment, liver of treated rats were assessed for histopathology. 99mTc-phytate uptake per gram tissue of the livers as %ID/g in control, solvent, MTX, Vit E, Vit E + MTX and MTX groups were 8.99% ± 1.37, 8.53% ± 2.91, 8.65% ± 3.84, 3.22% ± 1.09 and 8.38% ± 2.68. Vit E administration with MTX resulted in a significant increasing in the level of %ID/g. Vit E treatment improved the shape of live in planner image. Histophatological examinations showed a protective effect of Vit E against MTX-induced hepatoxicity in rats. The results showed that Vit E significantly attenuates the MTX-induced hepatotoxicity in rats, and 99mTc-phytate uptake in liver as well as liver image to be acceptable techniques for assessment of liver and spleen damages and/or their tissues protective effects in animal model.

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  1. Peters GJ, van der Wilt CL, van Moorsel CJ, et al. Basis for effective combination cancer chemotherapy with antimetabolites. Pharmacol Ther. 2000; 87(2-3): 227–253.
  2. Choudhury RC, Ghosh SK, Palo AK. Cytogenetic toxicity of methotrexate in mouse bone marrow. Environ Toxicol Pharmacol. 2000; 8(3): 191–196.
  3. Jensen SB, Mouridsen HT, Reibel J, et al. Adjuvant chemotherapy in breast cancer patients induces temporary salivary gland hypofunction. Oral Oncol. 2008; 44(2): 162–173.
  4. Sener G, Ekşioğlu-Demiralp E, Cetiner M, et al. L-Carnitine ameliorates methotrexate-induced oxidative organ injury and inhibits leukocyte death. Cell Biol Toxicol. 2006; 22(1): 47–60.
  5. Rang HP, Dale MM, Ritter JM, Flower RF, Henderson G. Rang & Dale's Pharmacology. 5th. Churchill Livingston, London 2003: 490–502.
  6. Thomas JA, Aithal GP. Monitoring liver function during methotrexate therapy for psoriasis: are routine biopsies really necessary? Am J Clin Dermatol. 2005; 6(6): 357–363.
  7. Caetano NN, Campello AP, Carnieri EG, et al. Effect of methotrexate (MTX) on NAD(P)+ dehydrogenases of HeLa cells: malic enzyme, 2-oxoglutarate and isocitrate dehydrogenases. Cell Biochem Funct. 1997; 15(4): 259–264, doi: 10.1002/(SICI)1099-0844(199712)15:4<259::AID-CBF749>3.0.CO;2-D.
  8. Uraz S, Tahan V, Aygun C, et al. Role of ursodeoxycholic acid in prevention of methotrexate-induced liver toxicity. Dig Dis Sci. 2008; 53(4): 1071–1077.
  9. Babiak RM, Campello AP, Carnieri EG, et al. Methotrexate: pentose cycle and oxidative stress. Cell Biochem Funct. 1998; 16(4): 283–293, doi: 10.1002/(SICI)1099-0844(1998120)16:4<283::AID-CBF801>3.0.CO;2-E.
  10. Armagan I, Bayram D, Candan IA, et al. Effects of pentoxifylline and alpha lipoic acid on methotrexate-induced damage in liver and kidney of rats. Environ Toxicol Pharmacol. 2015; 39(3): 1122–1131.
  11. Vardi N, Parlakpinar H, Cetin A, et al. Protective effect of beta-carotene on methotrexate-induced oxidative liver damage. Toxicol Pathol. 2010; 38(4): 592–597.
  12. Gulec M, Gurel A, Armutcu F. Vitamin E protects against oxidative damage caused by formaldehyde in the liver and plasma of rats. Mol Cell Biochem. 2006; 290(1-2): 61–67.
  13. Ozdil S, Bolkent S, Yanardag R, et al. Protective effects of ascorbic acid, dl-alpha-tocopherol acetate, and sodium selenate on ethanol-induced liver damage of rats. Biol Trace Elem Res. 2004; 97(2): 149–162.
  14. Groshar D, Slobodin G, Zuckerman E. Quantitation of liver and spleen uptake of (99m)Tc-phytate colloid using SPECT: detection of liver cirrhosis. J Nucl Med. 2002; 43(3): 312–317.
  15. Hoefs JC, Wang F, Kanel G, et al. The liver-spleen scan as a quantitative liver function test: correlation with liver severity at peritoneoscopy. Hepatology. 1995; 22(4 Pt 1): 1113–1121.
  16. Picard D, Infante-Rivard C, Villeneuve JP, et al. Extrahepatic uptake of technetium-99m-phytate: a prognostic index in patients with cirrhosis. J Nucl Med. 1990; 31(4): 436–440.
  17. Gonzalez P, Humeres P, Arroyo A, et al. Liver uptake with phytate colloid in normal subjects and patients with cirrhosis: a possible alternative to extrahepatic measurements—reply. J Nucl Med. 1991; 32(7): 1467–1468.
  18. Huet PM, Chartrand R, Marleau D. Extrahepatic uptake of 99mTc-phytate: its mechanism and significance in chronic liver disease. Gastroenterology. 1980; 78(1): 76–80.
  19. Gao Li, Yang F, Ren C, et al. Diagnosis of cirrhotic portal hypertension and compensatory circulation using transsplenic portal scintigraphy with (99m)Tc-phytate. J Nucl Med. 2010; 51(1): 52–56.
  20. Brunt EM. Grading and staging the histopathological lesions of chronic hepatitis: the Knodell histology activity index and beyond. Hepatology. 2000; 31(1): 241–246.
  21. Zhang C, Wang Y, Chen H, et al. Protective effect of the herbal medicine Gan‑fu‑kang against carbon tetrachloride‑induced liver fibrosis in rats. Mol Med Rep. 2013; 8(3): 954–962.
  22. Abolmaali SS, Tamaddon AM, Dinarvand R. A review of therapeutic challenges and achievements of methotrexate delivery systems for treatment of cancer and rheumatoid arthritis. Cancer Chemother Pharmacol. 2013; 71(5): 1115–1130.
  23. Hazlewood GS, Barnabe C, Tomlinson G, et al. Methotrexate monotherapy and methotrexate combination therapy with traditional and biologic disease modifying anti-rheumatic drugs for rheumatoid arthritis: A network meta-analysis. Cochrane Database Syst Rev. 2016; 353(8): CD010227.
  24. Puig L. Methotrexate: new therapeutic approaches. Actas Dermosifiliogr. 2014; 105(6): 583–589.
  25. Sotoudehmanesh R, Anvari B, Akhlaghi M, et al. Methotrexate hepatotoxicity in patients with rheumatoid arthritis. Middle East J Dig Dis. 2010; 2(2): 104–109.
  26. Barker J, Horn EJ, Lebwohl M, et al. International Psoriasis Council. Assessment and management of methotrexate hepatotoxicity in psoriasis patients: report from a consensus conference to evaluate current practice and identify key questions toward optimizing methotrexate use in the clinic. J Eur Acad Dermatol Venereol. 2011; 25(7): 758–764.
  27. Albrecht K, Müller-Ladner U. Side effects and management of side effects of methotrexate in rheumatoid arthritis. Clin Exp Rheumatol. 2010; 28(5 Suppl 61): S95–101.
  28. Mukherjee S, Ghosh S, Choudhury S, et al. Pomegranate reverses methotrexate-induced oxidative stress and apoptosis in hepatocytes by modulating Nrf2-NF-κB pathways. J Nutr Biochem. 2013; 24(12): 2040–2050.
  29. Dalaklioglu S, Genc GE, Aksoy NH, et al. Resveratrol ameliorates methotrexate-induced hepatotoxicity in rats via inhibition of lipid peroxidation. Hum Exp Toxicol. 2013; 32(6): 662–671.
  30. Ali N, Rashid S, Nafees S, et al. Beneficial effects of Chrysin against Methotrexate-induced hepatotoxicity via attenuation of oxidative stress and apoptosis. Mol Cell Biochem. 2014; 385(1-2): 215–223.
  31. Cure E, Kirbas A, Tumkaya L, et al. Protective effect of infliximab on methotrexate-induced liver injury in rats: unexpected drug interaction. J Cancer Res Ther. 2015; 11(1): 164–169.
  32. Hamazaki K, Sato S, Yunoki M, et al. Kupffer cell function in chronic liver injury and after partial hepatectomy. Res Exp Med (Berl). 1994; 194(4): 237–246.
  33. Watanabe T, Kondo M, Hirasa M, et al. Scintigraphic study of regenerative nodules due to fulminant hepatic failure. J Gastroenterol. 2003; 38(7): 695–699.
  34. Picard D, Infante-Rivard C, Villeneuve JP, et al. Extrahepatic uptake of technetium-99m-phytate: a prognostic index in patients with cirrhosis. J Nucl Med. 1990; 31(4): 436–440.
  35. Masui T, Yan H, Kosugi I, et al. Assessment of early radiation effects on the liver. Comparison of SPECT and MR. Acta Radiol. 1996; 37(5): 665–671.
  36. Niki E. Role of vitamin E as a lipid-soluble peroxyl radical scavenger: in vitro and in vivo evidence. Free Radic Biol Med. 2014; 66: 3–12.
  37. Tahan G, Aytac E, Aytekin H, et al. Vitamin E has a dual effect of anti-inflammatory and antioxidant activities in acetic acid-induced ulcerative colitis in rats. Can J Surg. 2011; 54(5): 333–338.
  38. Singh U, Devaraj S, Jialal I. Vitamin E, oxidative stress, and inflammation. Annu Rev Nutr. 2005; 25: 151–174.
  39. Jiang Q. Natural forms of vitamin E: metabolism, antioxidant, and anti-inflammatory activities and their role in disease prevention and therapy. Free Radic Biol Med. 2014; 72: 76–90.