Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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The Relationship between Prohibitin 1 Expression, Hepatotoxicity Induced by Acetaminophen, and Hepatoprotection by S-Adenosylmethionine in AML12 Cells

  • Eunhye Cho (Department of Nutritional Science and Food Management, Ewha Womans University) ;
  • Soohan Jung (Department of Integrated Biomedical and Life Science, Korea University) ;
  • Jina Kim (Department of Human Biology, University of Southern California) ;
  • Kwang Suk Ko (Department of Nutritional Science and Food Management, Ewha Womans University)
  • Received : 2022.07.18
  • Accepted : 2022.09.16
  • Published : 2022.11.28
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Abstract

Prohibitin 1 (Phb1) is a pleiotropic protein, located mainly in the mitochondrial inner membrane and involved in the regulation of cell proliferation and the stabilization of mitochondrial protein. Acetaminophen (APAP) is one of the most commonly used over-the-counter analgesics worldwide. However, at high dose, the accumulation of N-acetyl-p-benzoquinone imine (NAPQI) can lead to APAP-induced hepatotoxicity. In this study, we sought to understand the regulation of mRNA expression in relation to APAP and GSH metabolism by Phb1 in normal mouse AML12 hepatocytes. We used two different Phb1 silencing levels: high-efficiency (HE, >90%) and low-efficiency (LE, 50-60%). In addition, the siRNA-transfected cells were further pretreated with 0.5 mM of Sadenosylmethionine (SAMe) for 24 h before treatment with APAP at different doses (1-2 mM) for 24 h. The expression of APAP metabolism-related and antioxidant genes such as Cyp2e1 and Ugt1a1 were increased during SAMe pretreatment. Moreover, SAMe increased intracellular GSH concentration and it was maintained after APAP treatment. To sum up, Phb1 silencing and APAP treatment impaired the metabolism of APAP in hepatocytes, and SAMe exerted a protective effect against hepatotoxicity by upregulating antioxidant genes.

Keywords

Acknowledgement

This research was supported by the National Research Foundation of Korea (NRF) funded by the Ministry of Science and ICT. (NRF-2020R1A2C110245), and also by the BK21 FOUR (Fostering Outstanding Universities for Research) funded by the Ministry of Education (MOE, Korea) and National Research Foundation of Korea (NRF5199990614253). Also all authors thank to Samoh Pharm Co., Ltd to provide S-adenosylmethionine for this research.

References

  1. McGill MR, Jaeschke H. 2013. Metabolism and disposition of acetaminophen: recent advances in relation to hepatotoxicity and diagnosis. Pharm. Res. 30: 2174-2187. https://doi.org/10.1007/s11095-013-1007-6
  2. Khodayar MJ, et al. 2018. Betaine protects mice against acetaminophen hepatotoxicity possibly via mitochondrial complex II and glutathione availability. Biomed. Pharmacother. 103: 1436-1445. https://doi.org/10.1016/j.biopha.2018.04.154
  3. Roh T, et al. 2018. Detoxifying effect of pyridoxine on acetaminophen-induced hepatotoxicity via suppressing oxidative stress injury. Food. Chem. Toxicol. 114: 11-22. https://doi.org/10.1016/j.fct.2018.02.017
  4. Burcham PC, Harman AW. 1991. Acetaminophen toxicity results in site-specific mitochondrial damage in isolated mouse hepatocytes. J. Biol. Chem. 266: 5049-5054. https://doi.org/10.1016/S0021-9258(19)67754-9
  5. Bajt M, Knight TR, Lemasters JJ, Jaeschke H. 2004. Acetaminophen-induced oxidant stress and cell injury in cultured mouse hepatocytes: protection by N-acetyl cysteine. Toxicol. Sci. 80: 343-349. https://doi.org/10.1093/toxsci/kfh151
  6. Jaeschke H, McGill MR, Williams CD, Ramachandran A. 2011. Current issues with acetaminophen hepatotoxicity - a clinically relevant model to test the efficacy of natural products. Life Sci. 88: 737-745. https://doi.org/10.1016/j.lfs.2011.01.025
  7. Ramani K, Mavila N, Ko KS, Mato JM, Lu SC. 2016. Prohibitin 1 regulates the H19-Igf2 axis and proliferation in hepatocytes. J. Biol. Chem. 291: 24148-24159. https://doi.org/10.1074/jbc.M116.744045
  8. Xu Z, Wu J, Zha X. 2011. Up-regulation of prohibitin 1 is involved in the proliferation and migration of liver cancer cells. Sci. China. Life. Sci. 54: 121-127. https://doi.org/10.1007/s11427-010-4130-1
  9. Mishra S, Murphy LC, Nyomba BG, Murphy LJ. 2005. Prohibitin: a potential target for new therapeutics. Trends Mol. Med. 11: 192-197. https://doi.org/10.1016/j.molmed.2005.02.004
  10. Ko KS, Tomasi ML, Iglesias-Ara A, French BA, French SW, Ramani K, Lu SC. 2010. Liver-specific deletion of prohibitin 1 results in spontaneous liver injury, fibrosis, and hepatocellular carcinoma in mice. Hepatology 52: 2096-2108. https://doi.org/10.1002/hep.23919
  11. Anstee QM, Day CP. 2012. S-adenosylmethionine (SAMe) therapy in liver disease: a review of current evidence and clinical utility. J. Hepatol. 57: 1097-1109. https://doi.org/10.1016/j.jhep.2012.04.041
  12. Lu SC, Mato JM. 2008. S-Adenosylmethionine in cell growth, apoptosis and liver cancer. J. Gastroenterol. Hepatol. 23: S73-S77. https://doi.org/10.1111/j.1440-1746.2007.05289.x
  13. Lu SC. 2000. S-adenosylmethionine. Int. J. Biochem. Cell. Biol. 32: 391-395. https://doi.org/10.1016/S1357-2725(99)00139-9
  14. Mato JM, Martinez-Chantar ML, Lu SC. 2015. S-adenosylmethionine metabolism and liver disease. Ann. Hepatol. 12: 183-189.
  15. Gonzalez FJ. 2005. Role of cytochromes P450 in chemical toxicity and oxidative stress: studies with CYP2E1. Mutat. Res-Fund. Mol. M. 569: 101-110. https://doi.org/10.1016/j.mrfmmm.2004.04.021
  16. Lee SS, Buters JT, Pineau T, Fernandez-Salguero P, GonzalezFJ. 1996. Role of CYP2E1 in the hepatotoxicity of acetaminophen. J. Biol. Chem. 271: 12063-12067. https://doi.org/10.1074/jbc.271.20.12063
  17. Mackenzie PI, Bock KW, Burchell B, Guillemette C, Ikushiro SI, Iyanagi T, et al. 2005. Nomenclature update for the mammalian UDP glycosyltransferase (UGT) gene superfamily. Pharmacogenet. Genomics. 15: 677-685. https://doi.org/10.1097/01.fpc.0000173483.13689.56
  18. Tanikawa K, Torimura T. 2006. Studies on oxidative stress in liver diseases: important future trends in liver research. Med. Mol. Morphol. 39: 22-27. https://doi.org/10.1007/s00795-006-0313-z
  19. Zhu R, Wang Y, Zhang L, Guo Q. 2012. Oxidative stress and liver disease. Hepatol. Res. 42: 741-749. https://doi.org/10.1111/j.1872-034X.2012.00996.x
  20. Poss KD, Tonegawa S. 1997. Reduced stress defense in heme oxygenase 1-deficient cells. Proc. Natl. Acad. Sci. USA94: 10925-10930. https://doi.org/10.1073/pnas.94.20.10925
  21. Yamawaki H, Berk BC. 2006. Thioredoxin in the cardiovascular system. J. Mol. Med. 84: 997-1003. https://doi.org/10.1007/s00109-006-0109-6
  22. Jan YH, Heck DE, Dragomir AC, Gardner CR, Laskin DL, Laskin, JD. 2014. Acetaminophen reactive intermediates target hepatic thioredoxin reductase. Chem. Res. Toxicol. 27: 882-894. https://doi.org/10.1021/tx5000443
  23. Yan M, Huo Y, Yin S, Hu H. 2018. Mechanisms of acetaminophen-induced liver injury and its implications for therapeutic interventions. Redox. Biol. 17: 274-283. https://doi.org/10.1016/j.redox.2018.04.019
  24. Lee SM, Cho TS, Kim DJ, Cha YN. 1999. Protective effect of ethanol against acetaminophen-induced hepatotoxicity in mice: role of NADH: quinone reductase. Biochem. Pharmacol. 58: 1547-1555. https://doi.org/10.1016/S0006-2952(99)00248-8
  25. Lu SC. 1999. Regulation of hepatic glutathione synthesis: current concepts and controversies. FASEB J. 13: 1169-1183. https://doi.org/10.1096/fasebj.13.10.1169
  26. Behrends V, Giskeodegard GF, Bravo-Santano N, Letek M, Keun HC. 2019. Acetaminophen cytotoxicity in HepG2 cells is associated with a decoupling of glycolysis from the TCA cycle, loss of NADPH production, and suppression of anabolism. Arch. Toxicol. 93: 341-353. https://doi.org/10.1007/s00204-018-2371-0
  27. Kaplowitz N. 1981. The importance and regulation of hepatic glutathione. Yale J. Biol. Med. 54: 497.
  28. Franklin CC, Backos DS, Mohar I, White CC, Forman HJ, Kavanagh TJ. 2009. Structure, function, and post-translational regulation of the catalytic and modifier subunits of glutamate cysteine ligase. Mol. Aspects Med. 30: 86-98.  https://doi.org/10.1016/j.mam.2008.08.009