The Relationship between Prohibitin 1 Expression, Hepatotoxicity Induced by Acetaminophen, and Hepatoprotection by S-Adenosylmethionine in AML12 Cells
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
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.
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.
참고문헌 (28)
- McGill MR, "Metabolism and disposition of acetaminophen : recent advances in relation to hepatotoxicity and diagnosis", Pharm. Res, 30, pp.2174~2187 (2013)
- Khodayar MJ, "Betaine protects mice against acetaminophen hepatotoxicity possibly via mitochondrial complex II and glutathione availability", Biomed. Pharmacother, 103, pp.1436~1445 (2018)
- Roh T, "Detoxifying effect of pyridoxine on acetaminophen-induced hepatotoxicity via suppressing oxidative stress injury", Food. Chem. Toxicol, 114, pp.11~22 (2018)
- Burcham PC, "Acetaminophen toxicity results in site-specific mitochondrial damage in isolated mouse hepatocytes", J. Biol. Chem, 266, pp.5049~5054 (1991)
- Bajt M, "Acetaminophen-induced oxidant stress and cell injury in cultured mouse hepatocytes : protection by N-acetyl cysteine", Toxicol. Sci, 80, pp.343~349 (2004)
- Jaeschke H, "Current issues with acetaminophen hepatotoxicity-a clinically relevant model to test the efficacy of natural products", Life Sci, 88, pp.737~745 (2011)
- Ramani K, "Prohibitin 1 regulates the H19-Igf2 axis and proliferation in hepatocytes", J. Biol. Chem, 291, pp.24148~24159 (2016)
- Xu Z, "Up-regulation of prohibitin 1 is involved in the proliferation and migration of liver cancer cells", Sci. China. Life. Sci, 54, pp.121~127 (2011)
- Mishra S, "Prohibitin : a potential target for new therapeutics", Trends Mol. Med, 11, pp.192~197 (2005)
- Ko KS, "Liver-specific deletion of prohibitin 1 results in spontaneous liver injury, fibrosis, and hepatocellular carcinoma in mice", Hepatology, 52, pp.2096~2108 (2010)
- Anstee QM, "S-adenosylmethionine(SAMe)therapy in liver disease : a review of current evidence and clinical utility", J. Hepatol, 57, pp.1097~1109 (2012)
- Lu SC, "S-Adenosylmethionine in cell growth, apoptosis and liver cancer", J. Gastroenterol. Hepatol, 23, pp.73~77 (2008)
- Lu SC, "S-adenosylmethionine", Int. J. Biochem. Cell. Biol, 32, pp.391~395 (2000)
- Mato JM, "S-adenosylmethionine metabolism and liver disease", Ann. Hepatol, 12, pp.183~189 (2015)
- Gonzalez FJ, "Role of cytochromes P450 in chemical toxicity and oxidative stress : studies with CYP2E1", Mutat. Res-Fund. Mol. M, 569, pp.101~110 (2005)
- Lee SS, "Role of CYP2E1 in the hepatotoxicity of acetaminophen", J. Biol. Chem, 271, pp.12063~12067 (1996)
- Mackenzie PI, "Nomenclature update for the mammalian UDP glycosyltransferase(UGT)gene superfamily", Pharmacogenet. Genomics, 15, pp.677~685 (2005)
- Tanikawa K, "Studies on oxidative stress in liver diseases : important future trends in liver research", Med. Mol. Morphol, 39, pp.22~27 (2006)
- Zhu R, "Oxidative stress and liver disease", Hepatol. Res, 42, pp.741~749 (2012)
- Poss KD, "Reduced stress defense in heme oxygenase 1-deficient cells", Proc. Natl. Acad. Sci. USA, 94, pp.10925~10930 (1997)
- Yamawaki H, "Thioredoxin in the cardiovascular system", J. Mol. Med, 84, pp.997~1003 (2006)
- Yi-Hua Jan, "Acetaminophen Reactive Intermediates Target Hepatic Thioredoxin Reductase", Chemical Research in Toxicology, 27(5), pp.882~894 (2014), American Chemical Society (ACS)[DOI]
- Yan M, "Mechanisms of acetaminophen-induced liver injury and its implications for therapeutic interventions", Redox. Biol, 17, pp.274~283 (2018)
- Lee SM, "Protective effect of ethanol against acetaminophen-induced hepatotoxicity in mice : role of NADH : quinone reductase", Biochem. Pharmacol, 58, pp.1547~1555 (1999)
- Lu SC, "Regulation of hepatic glutathione synthesis : current concepts and controversies", FASEB J, 13, pp.1169~1183 (1999)
- Behrends V, "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, pp.341~353 (2019)
- Kaplowitz N, "The importance and regulation of hepatic glutathione", Yale J. Biol. Med, 54, pp.497~ (1981)
- Franklin CC, "Structure, function, and post-translational regulation of the catalytic and modifier subunits of glutamate cysteine ligase", Mol. Aspects Med, 30, pp.86~98 (2009)