Evaluation of the Efficacy of Glutathione Administration in Acetaminophen-Induced Hepatotoxicity in Experimental Rats

Subject Areas : Journal of Chemical Health Risks

Reham Nafad Elbendary ¹ , Ghada Hassabo ² , Abeer Mostafa ³ , Dina Sabry ⁴ , heba Abdelmonaem Ibrahim ⁵ , Amr Mohamed Tawfik Khattab ⁶

1 - lecturer of toxicology department Cairo univeristy
2 - toxicology department, Cairo university
3 - Cairo University
4 - Cairo University
5 - Cairo univeristy
6 - toxicology department, Cairo university

Received: 2020-10-04 Accepted : 2022-07-02 Published : 2024-03-16

Keywords: DNA damage, Liver function, GSH, Comet assay, APAP hepatotoxicity,

Abstract :

One of the most common causes of acute liver failure is acetaminophen overdose. The antidote N-acetylcysteine acts by scavenging the reactive metabolite, but its therapeutic limitation necessitates the development of additional therapeutic approaches that can benefit late-presenting patients. Glutathione (GSH) is the most abundant intracellular nonprotein thiol that has an important role in the regulation of many cellular physiologic functions such as redox-homeostatic buffering. This study aims to evaluate the efficacy of GSH supplementation in the recovery of deteriorated liver functions in induced acute acetaminophen toxicity rats; in addition to determining its value in the preservation of DNA integrity in such toxicity. This experimental study was done on 36 albino rats which were divided into three groups (n=12 rats / group) as follows, group1: Control group, group 2: Acetaminophen (APAP) treated group, group 3: APAP and glutathione treated group. Each group was subdivided into 2 subgroups (n=6) and they were sacrificed at 12 hours and 24 hours sequentially. The extent of hepatic inflammation, oxidative stress, and DNA damage was evaluated using histopathological study, and comet analysis, and biochemical markers (ALT, GSH, and MDA). GSH supplementation (APAP and glutathione treated group) significantly improved liver functions resulting in; a statistically significant decrease in ALT levels, reducing Malondialdehyde (MDA) levels, and preserving DNA integrity. GSH is a highly effective alternative in the treatment of APAP hepatotoxicity.

References:

1. Kisaoglu A., Borekci B., Yapca O.E., Bilen H., Suleyman H., 2013. Tissue damage and oxidant/antioxidant balance. Eurasian J Med. 45:47e9


2. Karihtala P., Soini Y., 2007. Reactive oxygen species and antioxidant mechanisms in human tissues and their relation to malignancies. APMIS. 115, 81e103.


3. Fakurazi S., Sharifudin S.A., Arulselvan P., 2012. Moringa oleifera hydroethanolic extracts effectively alleviate acetaminophen-induced hepatotoxicity in experimental rats through their antioxidant nature. Molecules, 17, 8334–50


4. Al-Belooshi T., John A., Al-Otaiba A., Raza H., 2010, Acetaminophen-induced mitochondrial oxidative stress in murine J774.2 monocyte macrophages. Am J Biomed Sci. 2,142–54


5. Fouad A.A., Al-Mulhim A.S., Jresat I., Gomaa W., 2012. Therapeutic role of telmisartan against acetaminophen hepatotoxicity in mice. Eur J Pharmacol. 693: 64–71


6. Kumari A., Kakkar P., 2012. Lupeol prevents acetaminophen-induced in vivo hepatotoxicity by altering the Bax/Bcl-2 and oxidative stress-mediated mitochondrial signaling cascade. Life Sci. 90, 561–70


7. Gujral J.S., Knight T.R., Farhood A., Bajt M.L., Jaeschke H., 2002. Mode of cell death after acetaminophen overdose in mice: apoptosis or oncotic necrosis? Toxicol Sci. 67(2), 322–328. doi: 10.1093/toxsci/67.2.322.


8. Kisaoglu A., Ozogul B., Turan M.I., Yilmaz I., Demiryilmaz I., Atamanalp S.S., Bakan E., Suleyman H., 2014. Damage induced by paracetamol compared with N-acetylcysteine. J Chin Med Assoc. 77(9), 463-8. doi: 10.1016/j.jcma.2014.01.011. Epub 2014 Jul 12. PMID: 25028290.


9. McGill M.R., Yan H.M., Ramachandran A., Murray G.J., Rollins D.E., Jaeschke H., 2011. A human model to study mechanisms of acetaminophen hepatotoxicity. J Hepatol. 53, 974–82


10. Wu G., Fang Y.Z., Yang S., Lupton J.R., Turner N.D., 2004. Glutathione metabolism and its implications for health. J Nutr. 134(3), 489-92. doi: 10.1093/jn/134.3.489.


11. Fakurazi S., Sharifudin S.A., Arulselvan P., 2012. Moringa oleifera hydroethanolic extracts effectively alleviate acetaminophen-induced hepatotoxicity in experimental rats through their antioxidant nature. Molecules. 17, 8334–50


12. Al-Belooshi T., John A., Al-Otaiba A., Raza H., 2010. Acetaminophen-induced mitochondrial oxidative stress in murine J774.2 monocyte macrophages. Am J Biomed Sci. 2, 142–54


13. Knight T.R., Jaeschke H., 2002. Acetaminophen-induced inhibition of Fas receptormediated liver cell apoptosis: mitochondrial dysfunction versus glutathione depletion. Toxicol Appl Pharmacol. 181, 133-141


14. James L.P., McCullough S.S., Lamps L.W., Hinson J.A., 2003. Effect of Nacetylcysteine on acetaminophen toxicity in mice: relationship to reactive nitrogen and cytokine formation. Toxicol Sci. 75, 458-467.


15. Bajt M.L., Knight T.R., Farhood F., Jaeschke H., 2003. Scavenging peroxynitrite with glutathione promotes regeneration and enhances survival during acetaminophen-induced liver injury in mice. J Pharmacol Exp Ther. 307, 67-73.


16. Knight T.R., HO Y.S., Farhood F., Jaeschke H., 2002. Peroxynitrite is a critical mediator of acetaminophen hepatotoxicity in murine liver: protection by glutathione. J Pharmacol Exp Ther. 303, 468-475.


17. Singh N.P., McCoy M.T., Tice R.R., Schneider E.L., 1988. A simple technique for quantitation of low levels of DNA damage in individual cells. Exp Cell Res. 175, 184-191


18. Chan Y., 2003. Biostatistics102: Quantitative Data – Parametric & Non-parametric Tests. Singapore Med J. 44, 391-396.


19. Ramachandran A., Jaeschke H., 2018. Acetaminophen Toxicity: Novel Insights Into Mechanisms and Future Perspectives. Gene Expr. 18(1), 19-30. doi: 10.3727/ 105221617X15084371374138. Epub 2017 Oct 20. Review. PMID:29054140


20. Floyd J.S., Barbehenn E., Lurie P., Wolfe S.M., 2009. Case series of liver failure associated with rosiglitazone and pioglitazone. Pharmacoepidemiol Drug Saf. 18, 1238-43


21. Uchida N.S., Silva-Filho S.E., Cardia G.F.E., Cremer E., Silva-Comar F.M.S., Silva E.L., Bersani-Amado C.A., Cuman R.K.N., 2017. Hepatoprotective Effect of Citral on Acetaminophen-Induced Liver Toxicity in Mice. Evid Based Complement Alternat Med. 2017, 1796209. doi: 10.1155/2017/1796209. Epub 2017 Jun 22. PMID: 28717379; PMCID: PMC5499238.


22. Valko M., Leibfritz D., Moncol J., Cronin M.T., Mazur M., Telser J., 2007. Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol. 39, 44e84.


23. Fakurazi S., Sharifudin S.A., Arulselvan P., 2012. Moringa oleifera hydroethanolic extracts effectively alleviate acetaminophen-induced hepatotoxicity in experimental rats through their antioxidant nature. Molecules.17, 8334–50


24. Al-Belooshi T., John A., Al-Otaiba A., Raza H., 2010.Acetaminophen-induced mitochondrial oxidative stress in murine J774.2 monocyte macrophages. Am J Biomed Sci. 2, 142–54


25. Fouad A.A., Al-Mulhim A.S., Jresat I., 2012 Gomaa W. Therapeutic role of telmisartan against acetaminophen hepatotoxicity in mice. Eur J Pharmacol. 693, 64–71


26. Kumari A., Kakkar P., 2012 Lupeol prevents acetaminophen-induced in vivo hepatotoxicity by altering the Bax/Bcl-2 and oxidative stress-mediated mitochondrial signaling cascade. Life Sci. 90, 561–70


27. Mitchell J.R., Jollow D.J., Potter W.Z., Gillette J.R., Brodie B.B., 1973. Acetaminophen-induced hepatic necrosis. IV. Protective role of glutathione. J Pharmacol Exp Ther. 187, 211–7


28. Choi J., Park K.H., Kim S.Z., Shin J.H., Jang S.I., 2013. The ameliorative effects of L-2-oxothiazolidine-4-carboxylate on acetaminophen-induced hepatotoxicity in mice. Molecules. 18, 3467–78


29. McGill M.R., Yan H.M., Ramachandran A., Murray G.J., Rollins D.E., Jaeschke H., 2011. HepaRG cells: A human model to study mechanisms of acetaminophen hepatotoxicity. J Hepatol. 53, 974–82


30. Fakurazi S., Sharifudin S.A., Arulselvan P., 2012. Moringa oleifera hydroethanolic extracts effectively alleviate acetaminophen-induced hepatotoxicity in experimental rats through their antioxidant nature. Molecules.17, 8334–50


31. Al-Belooshi T., John A., Al-Otaiba A., Raza H., 2010. Acetaminophen-induced mitochondrial oxidative stress in murine J774.2 monocyte macrophages. Am J Biomed Sci. 2,142–54


32. Qian L., Wang W., Zhou Y., Ma J., 2017. Effects of reduced glutathione therapy on chronic hepatitis B. Cent Eur J Immunol. 42(1), 97-100. doi: 10.5114/ceji.2016.65894.


33. More S.S., Nugent J., Vartak A.P., Nye S.M., Vince R., 2017. Hepatoprotective Effect of ψ-Glutathione in a Murine Model of Acetaminophen-Induced Liver Toxicity. Chem Res Toxicol. 30(3), 777-784. doi: 10.1021/acs.chemrestox.6b00291. Epub 2017 Feb 16. PMID: 28165728.


34. Knight T.R., Jaeschke H., 2002. Acetaminophen-induced inhibition of Fas receptormediated liver cell apoptosis: mitochondrial dysfunction versus glutathione depletion. Toxicol Appl Pharmacol. 181, 133-141.


35. Richard M.Green, Graham M., Michael R.O' Donovan, Kevin Chipman J., J. Hodges N., 2006. Subcellular compartmentalization of glutathione: Correlations with parameters of oxidative stress related to genotoxicity. Mutagenesis. 21(6), 383–390.


36. Morales A., Miranda M., Sánchez-Reyes A., Biete A., Fernández-Checa J.C., 1998. Oxidative damage of mitochondrial and nuclear DNA induced by ionizing radiation in human hepatoblastoma cells. Int J Radiat Oncol Biol Phys. 42(1), 191-203


37. McGill M.R., Sharpe M.R., Williams C.D., Taha M., Curry S.C., Jaeschke H., 2012. The mechanism underlying acetaminophen-induced hepatotoxicity in humans and mice involves mitochondrial damage and nuclear DNA fragmentation. J Clin Invest. 122(4), 1574-83. doi: 10.1172/JCI59755.