Randomized Controlled Trial
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World J Gastroenterol. Aug 7, 2014; 20(29): 10158-10165
Published online Aug 7, 2014. doi: 10.3748/wjg.v20.i29.10158
Cytoprotective effects of amifostine, ascorbic acid and N-acetylcysteine against methotrexate-induced hepatotoxicity in rats
Sami Akbulut, Hulya Elbe, Cengiz Eris, Zumrut Dogan, Gulten Toprak, Emrah Otan, Erman Erdemli, Yusuf Turkoz
Sami Akbulut, Cengiz Eris, Emrah Otan, Liver Transplant Institute, Inonu University Faculty of Medicine, 44280 Malatya, Turkey
Hulya Elbe, Department of Histology and Embriology, Inonu University Faculty of Medicine, 44280 Malatya, Turkey
Zumrut Dogan, Department of Anatomy, Adiyaman University Faculty of Medicine, 02100 Adiyaman, Turkey
Gulten Toprak, Department of Biochemistry, Dicle University Faculty of Medicine, 21280 Diyarbakir, Turkey
Erman Erdemli, Yusuf Turkoz, Department of Biochemistry, Inonu University Faculty of Medicine, 44280 Malatya, Turkey
Author contributions: Akbulut S, Eris C, Dogan Z and Erdemli E performed all of experimental design and surgical procedures; Akbulut S, Elbe H and Otan E contributed to writing the paper and review of the literature; Elbe H provided histopathologic information and performed statistical analysis; Toprak G and Turkoz Y provided biochemical information.
Correspondence to: Sami Akbulut, Assistant Professor, FICS, FACS, Liver Transplant Institute, Inonu University Faculty of Medicine, 44280 Malatya, Turkey. akbulutsami@gmail.com
Telephone: +90-422-3410660 Fax: +90-422-3410036
Received: December 9, 2013
Revised: February 11, 2014
Accepted: April 8, 2014
Published online: August 7, 2014
Processing time: 240 Days and 16.2 Hours
Abstract

AIM: 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].

CONCLUSION: 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.

Keywords: Methotrexate; Amifostine; N-acetyl cysteine; Ascorbic acid; Hepatotoxicity; Oxidative stress

Core tip: While the underlying mechanism of methotrexate (MTX)-induced hepatotoxicity remains to be fully elucidated, increases in oxidative stress have been linked to the effects of MTX; moreover, MTX-induced toxicity has been shown to be associated with increases in lipid peroxidation in various tissues, such as liver, using rat model systems. A few previous studies have shown that prophylactic delivery of antioxidant agents can prevent MTX-induced hepatotoxicity. Based on this information, we aimed to investigate the role of oxidative stress in MTX-induced hepatotoxicity and to evaluate the potential therapeutic effects of the cytoprotective antioxidants N-acetylcysteine, amifostine, and ascorbic acid.