Protective Effects of Essential Oils as Natural Antioxidants against Hepatotoxicity Induced by Cyclophosphamide in Mice
Authors: Lobna S El-Hosseiny
Munther A Nashashibi
Salah A Sheweita
Journal: PLoS ONE (2016)
Discipline: Multidisciplinary
Keywords: cyclophosphamide; hepatotoxicity; essential oils; antioxidants; liver
Affiliations: Department of Biotechnology, Institute of Graduate Studies and Research, Alexandria University, Alexandria, Egypt., Department of Environmental Studies, Institute of Graduate Studies and Research, Alexandria University, Alexandria, Egypt.

Abstract:  Clinical application of cyclophosphamide (CP) as an anticancer drug is often limited due to its toxicity. CP is metabolized mainly in the liver by cytochrome P450 system into acrolein which is the proximate toxic metabolite. Many different natural antioxidants were found to alleviate the toxic effects of various toxic agents via different mechanisms. Therefore, the present study aimed at investigating the role of essential oils extracted from fennel, cumin and clove as natural antioxidants in the alleviation of hepatotoxicity induced by CP through assessment of hepatotoxicity biomarkers (AST, ALT, ALP), histopathology of liver tissues as well as other biochemical parameters involved in the metabolism of CP. The data of the present study showed that treatment of male mice with cyclophosphamide (2.5 mg/Kg BW) as repeated dose for 28 consecutive days was found to induce hepatotoxicity through the elevation in the activities of AST, ALT, and ALP. Combined administration of any of these oils with CP to mice partially normalized the altered hepatic biochemical markers caused by CP, whereas administration of fennel, clove or cumin essential oils alone couldn't change liver function indices. Moreover, CP caused histological changes in livers of mice including swelling and dilation in sinusoidal space, inflammation in portal tract and hepatocytes, as well as, hyperplasia in Kuppfer cells. However, co-administration of any of the essential oils with CP alleviated to some extent the changes caused by CP but not as the normal liver. CP was also found to induce free radical levels (measured as thiobarbituric acid reactive substances) and inhibited the activities of superoxide dismutase, glutathione reductase, and catalase as well as activities and protein expressions of both glutathione S-transferase (GSTπ) and glutathione peroxidase. Essential oils restored changes in activities of antioxidant enzymes (SOD, CAT, GR, GST, and GPx) caused by CP to their normal levels compared to control group. In addition, treatment of mice with CP was found to induce the protein expression of CYP 3A4, 2B1/2, 2C6, 2C23. Moreover, the present study showed that essential oils reduced the expression of CYPs 2E1, 3A4 but could not restore the expression of CYP 2C6 and 2C23 compared to CP-treated mice. Interestingly, pretreatment of mice with essential oil of clove was found to restore activities of DMN-dI, AHH, and ECOD which were induced by CP to their normal control levels. It is concluded that EOs showed a marked hepatoprotective effect against hepatotoxicity induced by CP. In addition, co-administration of CP with any of these oils might be used as a new strategy for cancer treatment to alleviate the hepatotoxicity induced by CP. 

Retracted: No

PubPeer comments:
 Figure 3 may have some repeated elements (with adjustment to dimensions) and evidence of splicing (arrow). Could the authors please post an original uncropped blot? I did not check elsewhere. *** Placed adjacently for easier comparison. ![file] *** Showing original placement. ![file] , 
 Figure 4 appears to have an image with two regions showing unexpected similarities, although they are not identical. *** ![file] , 
 Some additional problems spotted in Figure 4. ![file] , 
 No changes from original gels . please send me your email to send them. It seems you have more .time. This paper has been reviewed by three reviwers. please stop what you are doing since your comments are not based on any scientific bases. , 
 The b-actin blots used in Figures 2 and 3 also appear to have been used in several other papers by the same author(s). Same blot spotted in the following papers (marked with red boxes): * Salah A. Sheweita et al., PLOS ONE (2016), doi:10.1371 journal.pone.0165667 (this paper) ![file] * Salah A. Sheweita et al.,Oxidative Medicine and Cellular Longevity (2016), doi: 10.1155 2016 4970906, https: pubpeer.com publications 7EB7E48FB7C6AB67CC4A457A86581F ![file] * Salah A. Sheweita et al., PLOS ONE (2018), doi:10.1371 journal.pone.0202110, https: pubpeer.com publications EC715E36CC25E84B60A1664B821398 ![file] * Salah A. Sheweita et al., PLOS ONE (2020), doi:10.1371 journal.pone.0241509, https: pubpeer.com publications 2AEA8222A001DABDF861C1924D47B2 ![file] A close up of the use of the blot in these four papers: ![file] , 
 Dear Dr. Sheweita, if you have original gels to share, please share them here. No matter how many reviewers have looked at your paper, I am a scientist who has concerns about these blots. I am asking you to share the originals here. , 
 The authors attribute some of the effects seen in this paper to quercetin - however in a standard distilled essential oil there is likely none of this compound present as it is not a volatile substance whatsoever. , 
 #5 I sympathize with the authoru2019s viewpoint but remind the author that every paper that has been corrected or retracted after publication was also peer reviewed before publication. It is not a foolproof process. Could the authors please provide the original uncropped scans? https: www.nature.com nature-portfolio editorial-policies image-integrity#electrophoretic-gels-and-blots describes some best practices regarding image integrity and standards for gels and blots. External sites for larger files can be linked here. Some options: http: figshare.com http: Imgur.com https: www.google.com drive Baidu (u767eu5ea6u7f51u76d8) Tencent (u817eu8bafu5faeu4e91) ,