title: Cytoprotective effects of amifostine, ascorbic acid and N-acetylcysteine against methotrexate-induced hepatotoxicity in rats.
abstract: OBJECTIVE
  To investigate the potential role of oxidative stress and the possible therapeutic effects of N-acetyl cysteine (NAC), amifostine (AMF) and ascorbic acid (ASC) in methotrexate (MTX)-induced hepatotoxicity.
  
  
  METHODS
  An MTX-induced hepatotoxicity model was established in 44 male Sprague Dawley rats by administration of a single intraperitoneal injection of 20 mg/kg MTX. Eleven of the rats were left untreated (Model group; n = 11), and the remaining rats were treated with a 7-d course of 50 mg/kg per day NAC (MTX + NAC group; n = 11), 50 mg/kg per single dose AMF (MTX + AMF group; n = 11), or 10 mg/kg per day ASC (MTX + ASC group; n = 11). Eleven rats that received no MTX and no treatments served as the negative control group. Structural and functional changes related to MTX- and the various treatments were assessed by histopathological analysis of liver tissues and biochemical assays of malondialdehyde (MDA), superoxide dismutase (SOD), catalase, glutathione (GSH) and xanthine oxidase activities and of serum levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and total bilirubin.
  
  
  RESULTS
  Exposure to MTX caused structural and functional hepatotoxicity, as evidenced by significantly worse histopathological scores [median (range) injury score: control group: 1 (0-3) vs 7 (6-9), P = 0.001] and significantly higher MDA activity [409 (352-466) nmol/g vs 455.5 (419-516) nmol/g, P < 0.05]. The extent of MTX-induced perturbation of both parameters was reduced by all three cytoprotective agents, but only the reduction in hepatotoxicity scores reached statistical significance [4 (3-6) for NAC, 4.5 (3-5) for AMF and 6 (5-6) for ASC; P = 0.001, P = 0.001 and P < 0.005 vs model group respectively]. Exposure to MTX also caused a significant reduction in the activities of GSH and SOD antioxidants in liver tissues [control group: 3.02 (2.85-3.43) μmol/g and 71.78 (61.88-97.81) U/g vs model group: 2.52 (2.07-3.34) μmol/g and 61.46 (58.27-67.75) U/g, P < 0.05]; however, only the NAC treatment provided significant increases in these antioxidant enzyme activities [3.22 (2.54-3.62) μmol/g and 69.22 (61.13-100.88) U/g, P < 0.05 and P < 0.01 vs model group respectively].
  
  
  CONCLUSIONS
  MTX-induced structural and functional damage to hepatic tissues in rats may involve oxidative stress, and cytoprotective agents (NAC > AMF > ASC) may alleviate MTX hepatotoxicity.
authors.affiliation: Sami Akbulut, Cengiz Eris, Emrah Otan, Liver Transplant Institute, Inonu University Faculty of Medicine, 44280 Malatya, Turkey.
authors.forename: Sami
authors.initials: S
authors.lastname: Akbulut
authors.affiliation: Sami Akbulut, Cengiz Eris, Emrah Otan, Liver Transplant Institute, Inonu University Faculty of Medicine, 44280 Malatya, Turkey.
authors.forename: Hulya
authors.initials: H
authors.lastname: Elbe
authors.affiliation: Sami Akbulut, Cengiz Eris, Emrah Otan, Liver Transplant Institute, Inonu University Faculty of Medicine, 44280 Malatya, Turkey.
authors.forename: Cengiz
authors.initials: C
authors.lastname: Eris
authors.affiliation: Sami Akbulut, Cengiz Eris, Emrah Otan, Liver Transplant Institute, Inonu University Faculty of Medicine, 44280 Malatya, Turkey.
authors.forename: Zumrut
authors.initials: Z
authors.lastname: Dogan
authors.affiliation: Sami Akbulut, Cengiz Eris, Emrah Otan, Liver Transplant Institute, Inonu University Faculty of Medicine, 44280 Malatya, Turkey.
authors.forename: Gulten
authors.initials: G
authors.lastname: Toprak
authors.affiliation: Sami Akbulut, Cengiz Eris, Emrah Otan, Liver Transplant Institute, Inonu University Faculty of Medicine, 44280 Malatya, Turkey.
authors.forename: Emrah
authors.initials: E
authors.lastname: Otan
authors.affiliation: Sami Akbulut, Cengiz Eris, Emrah Otan, Liver Transplant Institute, Inonu University Faculty of Medicine, 44280 Malatya, Turkey.
authors.forename: Erman
authors.initials: E
authors.lastname: Erdemli
authors.affiliation: Sami Akbulut, Cengiz Eris, Emrah Otan, Liver Transplant Institute, Inonu University Faculty of Medicine, 44280 Malatya, Turkey.
authors.forename: Yusuf
authors.initials: Y
authors.lastname: Turkoz
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology*; D004999:Amifostine / Q000494:pharmacology*; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D001205:Ascorbic Acid / Q000494:pharmacology*; D015415:Biomarkers / Q000378:metabolism; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D019610:Cytoprotection; D004195:Disease Models, Animal; D008099:Liver / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008297:Male; D008727:Methotrexate; D018384:Oxidative Stress / Q000187:drug effects*; D017207:Rats, Sprague-Dawley; D013997:Time Factors
chemical_list: D000975:Antioxidants; D015415:Biomarkers; D004999:Amifostine; D001205:Ascorbic Acid; D000111:Acetylcysteine; D008727:Methotrexate
keywords: Amifostine; Ascorbic acid; Hepatotoxicity; Methotrexate; N-acetyl cysteine; Oxidative stress
references.citation: Med Sci Monit. 2012 Dec;18(12):BR475-81
references.pmid: 23197226
references.citation: Curr Med Chem. 2004 Apr;11(8):1041-64
references.pmid: 15078165
references.citation: J Egypt Natl Canc Inst. 2008 Jun;20(2):141-8
references.pmid: 20029470
references.citation: J Pharm Pharm Sci. 2010;13(2):303-10
references.pmid: 20816014
references.citation: ScientificWorldJournal. 2012;2012:987508
references.pmid: 22566785
references.citation: Clin Biochem. 2005 Dec;38(12):1103-11
references.pmid: 16214125
references.citation: J Pharmacol Pharmacother. 2012 Jul;3(3):248-53
references.pmid: 23129960
references.citation: Ren Fail. 2005;27(6):771-3
references.pmid: 16350832
references.citation: Scand J Gastroenterol. 2004 Nov;39(11):1119-27
references.pmid: 15545171
references.citation: Cell Biochem Funct. 1998 Dec;16(4):283-93
references.pmid: 9857491
references.citation: Arch Biochem Biophys. 1959 May;82(1):70-7
references.pmid: 13650640
references.citation: Med Sci Monit. 2006 Aug;12(8):BR274-8
references.pmid: 16865059
references.citation: Dig Dis Sci. 2008 Apr;53(4):1071-7
references.pmid: 17934844
references.citation: J Lab Clin Med. 1967 Jul;70(1):158-69
references.pmid: 6066618
references.citation: Basic Clin Pharmacol Toxicol. 2009 Jul;105(1):1-9
references.pmid: 19371262
references.citation: Toxicol Pathol. 2010 Jun;38(4):592-7
references.pmid: 20448084
references.citation: Clin Biochem. 2004 Feb;37(2):112-9
references.pmid: 14725941
references.citation: Gerontology. 2001 Jul-Aug;47(4):186-8
references.pmid: 11408722
references.citation: Hum Exp Toxicol. 2013 Jun;32(6):662-71
references.pmid: 23424212
references.citation: Mol Cell Biochem. 2014 Jan;385(1-2):215-23
references.pmid: 24154663
references.citation: Anal Biochem. 1978 May;86(1):271-8
references.pmid: 655387
references.citation: Cell Biochem Funct. 2004 May-Jun;22(3):169-78
references.pmid: 15124182
references.citation: Clin Chem. 1988 Mar;34(3):497-500
references.pmid: 3349599
references.citation: Balkan Med J. 2013 Mar;30(1):99-104
references.pmid: 25207078