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Published Online: 10 February 2017

Antcin H Protects Against Acute Liver Injury Through Disruption of the Interaction of c-Jun-N-Terminal Kinase with Mitochondria

Publication: Antioxidants & Redox Signaling
Volume 26, Issue Number 5


Aim: Antrodia Camphorate (AC) is a mushroom that is widely used in Asian countries to prevent and treat various diseases, including liver diseases. However, the active ingredients that contribute to the biological functions remain elusive. The purpose of the present study is to test the hepatoprotective effect of Antcin H, a major triterpenoid chemical isolated from AC, in murine models of acute liver injury.
Results: We found that Antcin H pretreatment protected against liver injury in both acetaminophen (APAP) and galactosamine/tumor necrosis factor (TNF)α models. More importantly, Antcin H also offered a significant protection against acetaminophen-induced liver injury when it was given 1 h after acetaminophen. The protection was verified in primary mouse hepatocytes. Antcin H prevented sustained c-Jun-N-terminal kinase (JNK) activation in both models. We excluded an effect of Antcin H on acetaminophen metabolism and TNF receptor signaling and excluded a direct effect as a free radical scavenger or JNK inhibitor. Since the sustained JNK activation through its interaction with mitochondrial Sab, leading to increased mitochondrial reactive oxygen species (ROS), is pivotal in both models, we examined the effect of Antcin H on p-JNK binding to mitochondria and impairment of mitochondrial respiration. Antcin H inhibited the direct effect of p-JNK on isolated mitochondrial function and binding to isolated mitochondria.
Innovation and Conclusion: Our study has identified Antcin H as a novel active ingredient that contributes to the hepatoprotective effect of AC, and Antcin H protects against liver injury through disruption of the binding of p-JNK to Sab, which interferes with the ROS-dependent self-sustaining activation of MAPK cascade. Antioxid. Redox Signal. 26, 207–220.

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Abdelmegeed MA, Jang S, Banerjee A, Hardwick JP, and Song BJ. Robust protein nitration contributes to acetaminophen-induced mitochondrial dysfunction and acute liver injury. Free Radical Bio Med 60: 211–222, 2013.
Amir M, Liu K, Zhao E, and Czaja MJ. Distinct functions of JNK and c‐Jun in oxidant‐induced hepatocyte death. J Cell Biochem 113: 3254–3265, 2012.
An L, Wang X, and Cederbaum AI. Cytokines in alcoholic liver disease. Arch Toxicol 86: 1337–1348, 2012.
Chambers JW and LoGrasso PV. Mitochondrial c-Jun N-terminal kinase (JNK) signaling initiates physiological changes resulting in amplification of reactive oxygen species generation. J Biol Chem 286: 16052–16062, 2011.
Chen CH, Yang SW, and Shen YC. New steroid acids from Antrodia cinnamomea, a fungal parasite of Cinnamomum micranthum. J Nat Prod 1655–1661, 1995.
Coen M. Metabolic phenotyping applied to pre-clinical and clinical studies of acetaminophen metabolism and hepatotoxicity. Drug Metab Rev 47: 29–44, 2015.
Davidson DG and Eastham WN. Acute liver necrosis following overdose of paracetamol. Br Med J 2: 497–499, 1966.
Feng G and Kaplowitz N. Colchicine protects mice from the lethal effect of an agonistic anti-Fas antibody. J Clin Invest 105: 329–339, 2000.
Feng G and Kaplowitz N. Mechanism of staurosporine-induced apoptosis in murine hepatocytes. Am J Physiol-Gastrointest Liver Physiol 282: G825–G834, 2002.
Geethangili M and Tzeng YM. Review of pharmacological effects of Antrodia camphorata and its bioactive compounds. Evid Based Complement Alternat Med 212641, 2011.
Gulmez SE, Larrey D, Pageaux GP, Bernuau J, Bissoli F, Horsmans Y, Thorburn D, McCormick PA, Stricker B, Toussi M, Lignot-Maleyran S, Micon S, Hamoud F, Lassalle R, Jové J, Blin P, and Moore N. Liver transplant associated with paracetamol overdose: results from the seven‐country SALT study. Br J Clin Pharmacol 80: 599–606, 2015.
Gunawan BK, Liu ZX, Han D, Hanawa N, Gaarde WA, and Kaplowitz N. c-Jun N-terminal kinase plays a major role in murine acetaminophen hepatotoxicity. Gastroenterology 131: 165–178, 2006.
Han D, Shinohara M, Ybanez MD, Saberi B, and Kaplowitz N. Signal transduction pathways involved in drug-induced liver injury. Adverse Drug React 267–310, 2010.
Han D, Ybanez MD, Ahmadi S, Yeh K, and Kaplowitz N. Redox regulation of tumor necrosis factor signaling. Antioxid Redox Signal 11: 2245–2263, 2009.
Hanawa N, Shinohara M, Saberi B, Gaarde WA, Han D, and Kaplowitz N. Role of JNK translocation to mitochondria leading to inhibition of mitochondria bioenergetics in acetaminophen-induced liver injury. J Biol Chem 283: 13565–13577, 2008.
Hinson JA, Michael SL, Ault SG, and Pumford NR. Western blot analysis for Nitrotyrosine protein adducts in livers of saline-treated and acetaminophen-treated mice. Toxicol Sci 53: 467–473, 2000.
Hinson JA, Pike SL, Pumford NR, and Mayeux PR. Nitrotyrosine-protein adducts in hepatic centrilobular areas following toxic doses of acetaminophen in mice. Chem Res Toxicol 11: 604–607, 1998.
James LP, Mayeux PR, and Hinson JA. Acetaminophen-induced hepatotoxicity. Drug Metab Dispos 31: 1499–1506, 2003.
Ker YB, Peng CC, Chang WL, Chyau CC, and Peng RY. Hepatoprotective bioactivity of the glycoprotein, antrodan, isolated from Antrodia cinnamomea mycelia. PLoS One 9: e93191, 2014.
Kharbanda S, Saxena S, Yoshida K, Pandey P, Kaneki M, Wang Q, Cheng K, Chen YN, Campbell A, Sudha T, Yuan ZM, Narula J, Weichselbaum R, Nalin C, and Kufe D. Translocation of SAPK/JNK to mitochondria and interaction with Bcl-xL in response to DNA damage. J Biol Chem 275: 322–327, 2000.
Knight TR, Ho YS, Farhood A, and Jaeschke H. Peroxynitrite is a critical mediator of acetaminophen hepatotoxicity in murine livers: protection by glutathione. J Pharmacol Exp Ther 303: 468–475, 2002.
Larson AM, Polson J, Fontana RJ, Davern TJ, Lalani E, Hynan LS, Reisch JS, Schiødt FV, Ostapowicz G, Shakil AO, and Lee WM. Acetaminophen‐induced acute liver failure: results of a United States multicenter, prospective study. Hepatology 42: 1364–1372, 2005.
Lee WM, Squires RH, Nyberg SL, Doo E, and Hoofnagle JH. Acute liver failure: summary of a workshop. Hepatology 47: 1401–1415, 2008.
Liu YW, Lu KH, Ho CT, and Sheen LY. Protective effects of Antrodia cinnamomea against liver injury. J Tradit Complement Med 2: 284, 2012.
Lu MC, El-Shazly M, Wu TY, Du YC, Chang TT, Chen CF, Hsu YM, Lai KH, Chiu CP, Chang FR, and Wu YC. Recent research and development of Antrodia cinnamomea. Pharmacol Ther 139: 124–156, 2013.
Male KB, Rao YK, Tzeng YM, Montes J, Kamen A, and Luong JH. Probing inhibitory effects of Antrodia camphorata isolates using insect cell-based impedance spectroscopy: inhibition vs chemical structure. Chem Res Toxicol 21: 2127–2133, 2008.
McGill MR, Lebofsky M, Norris HRK, Slawson MH, Bajt ML, Xie Y, Williams CD, Wilkins DG, Rollins DE, and Jaeschke H. Plasma and liver acetaminophen-protein adduct levels in mice after acetaminophen treatment: dose–response, mechanisms, and clinical implications. Toxicol Appl Pharmacol 269: 240–249, 2013.
Nagai H, Matsumaru K, Feng G, and Kaplowitz N. Reduced glutathione depletion causes necrosis and sensitization to tumor necrosis factor‐α-induced apoptosis in cultured mouse hepatocytes. Hepatology 36: 55–64, 2002.
Nakagawa H, Maeda S, Hikiba Y, Ohmae T, Shibata W, Yanai A, Sakamoto K, Ogura K, Noguchi T, Karin M, Ichijo H, and Omata M. Deletion of apoptosis signal-regulating kinase 1 attenuates acetaminophen-induced liver injury by inhibiting c-Jun N-terminal kinase activation. Gastroenterology 135: 1311–1321, 2008.
Phuong DT, Ma CM, Hattori M, and Jin JS. Inhibitory effects of antrodins A-E from Antrodia cinnamomea and their metabolites on hepatitis C virus protease. Phytother Res 23: 582–584, 2009.
Qiao X, An R, Huang Y, Ji S, Li L, Tzeng YM, Guo DA, and Ye M. Separation of 25R/S-ergostane triterpenoids in the medicinal mushroom Antrodia camphorata using analytical supercritical-fluid chromatography. J Chromatogr A 1358: 252–260, 2014.
Qiao X, Wang Q, Ji S, Huang Y, Liu KD, Zhang ZX, Bo T, Tzeng YM, Guo DA, and Ye M. Metabolites identification and multi-component pharmacokinetics of ergostane and lanostanetriterpenoids in the anticancer mushroom Antrodiacinnamomea. J Pharm Biomed Anal 111: 266–276, 2015.
Rahman I, Kode A, and Biswas SK. Assay for quantitative determination of glutathione and glutathione disulfide levels using enzymatic recycling method. Nat Protoc 1: 3159–3165, 2006.
Saito C, Lemasters JJ, and Jaeschke H. c-Jun N-terminal kinase modulates oxidant stress and peroxynitrite formation independent of inducible nitric oxide synthase in acetaminophen hepatotoxicity. Toxicol Appl Pharmacol 246: 8–17, 2010.
Shen YC, Wang YH, Chou YC, Chen CF, Lin LC, Chang TT, Tien JH, and Chou CJ. Evaluation of the anti-inflammatory activity of zhankuic acids isolated from the fruiting bodies of Antrodia camphorata. Planta Med 70: 310–314, 2004.
Tahir M, Rehman MU, Lateef A, Khan R, Khan AQ, Qamar W, Ali F, O'Hamiza O, and Sultana S. Diosmin protects against ethanol-induced hepatic injury via alleviation of inflammation and regulation of TNF-α and NF-κB activation. Alcohol 47: 131–139, 2013.
Than TA, Win S, and Kaplowitz N. In vitro assays of mitochondrial function/dysfunction. Clin Pharmacol Ther 96: 665–668, 2014.
Tilg H and Diehl AM. Cytokines in alcoholic and nonalcoholic steatohepatitis. New Engl J Med 343: 1467–1476, 2000.
Tsou TC, Tsai FY, Wu MC, and Chang LW. The protective role of NF-κB and AP-1 in arsenite-induced apoptosis in aortic endothelial cells. Toxicol Appl Pharmacol 191: 177–187, 2003.
Twumasi‐Boateng K, Wang TW, Tsai L, Lee KH, Salehpour A, Bhat S, Tan MW, and Shapira M. An age-dependent reversal in the protective capacities of JNK signaling shortens Caenorhabditis elegans lifespan. Aging cell 11: 659–667, 2012.
Weston CR and Davis RJ. The JNK signal transduction pathway. Curr Opin Cell Biol 19: 142–149, 2007.
Win S, Than TA, Fernandez-Checa JC, and Kaplowitz N. JNK interaction with Sab mediates ER stress induced inhibition of mitochondrial respiration and cell death. Cell Death Dis 5: e989, 2014.
Win S, Than TA, Han D, Petrovic LM, and Kaplowitz N. c-Jun N-terminal kinase (JNK)-dependent acute liver injury from acetaminophen or tumor necrosis factor (TNF) requires mitochondrial Sab protein expression in mice. J Biol Chem 286: 35071–35078, 2011.
Win S, Than TA, Le BH, García-Ruiz C, Fernandez-Checa JC, and Kaplowitz N. Sab (Sh3bp5) dependence of JNK mediated inhibition of mitochondrial respiration in palmitic acid induced hepatocyte lipotoxicity. J Hepatol 62: 1367–1374, 2015.
Win S, Than TA, Min RWM, Aghajan M, and Kaplowitz N. JNK mediates mouse liver injury through a novel Sab (SH3BP5) dependent pathway leading to inactivation of intramitochondrial Src. Hepatology 63: 1987–2003, 2016.
Yang L, Sun G, Guo Y, Hou Z, and Chen S. Holistic evaluation of quality consistency of Ixeris sonchifolia (Bunge) Hance injectables by quantitative fingerprinting in combination with antioxidant activity and chemometric methods. PLoS One 11: e0148878, 2016.
Ye X, Liu SF, and Liu G. NF-κB to Ap-1 switch: a mechanism regulating the transition from endothelial barrier injury to barrier repair following endotoxemic lung injury. Lung 1: 2, 2014.

Information & Authors


Published In

cover image Antioxidants & Redox Signaling
Antioxidants & Redox Signaling
Volume 26Issue Number 5February 10, 2017
Pages: 207 - 220
PubMed: 27596680


Published in print: February 10, 2017
Published online: 10 February 2017
Published ahead of print: 11 October 2016
Published ahead of production: 5 September 2016
Accepted: 5 September 2016
Revision received: 11 August 2016
Received: 14 July 2016


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Yazhen Huo
Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, China.
USC Research Center for Liver Disease, Keck School of Medicine, University of Southern California, Los Angeles, California.
Sanda Win
USC Research Center for Liver Disease, Keck School of Medicine, University of Southern California, Los Angeles, California.
Tin Aung Than
USC Research Center for Liver Disease, Keck School of Medicine, University of Southern California, Los Angeles, California.
Shutao Yin
Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, China.
Min Ye
State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing, China.
Hongbo Hu
Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, China.
Neil Kaplowitz
USC Research Center for Liver Disease, Keck School of Medicine, University of Southern California, Los Angeles, California.


Address correspondence to:Prof. Hongbo HuBeijing Advanced Innovation Center for Food Nutrition and Human HealthCollege of Food Science and Nutritional EngineeringChina Agricultural UniversityNo. 17 Qinghua East RoadHaidian DistrictBeijing 100083China
E-mail: [email protected]
Dr. Neil KaplowitzUSC Research Center for Liver DiseaseKeck School of MedicineUniversity of Southern California2011 Zonal Avenue, HMR101Los Angeles, CA 90033E-mail: [email protected]

Author Disclosure Statement

No competing financial interests exist.

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