All Title Author
Keywords Abstract

PLOS ONE  2013 

Zidovudine (AZT) and Hepatic Lipid Accumulation: Implication of Inflammation, Oxidative and Endoplasmic Reticulum Stress Mediators

DOI: 10.1371/journal.pone.0076850

Full-Text   Cite this paper   Add to My Lib


The clinical effectiveness of Zidovudine (AZT) is constrained due to its side-effects including hepatic steatosis and toxicity. However, the mechanism(s) of hepatic lipid accumulation in AZT-treated individuals is unknown. We hypothesized that AZT-mediated oxidative and endoplasmic reticulum (ER) stress may play a role in the AZT-induced hepatic lipid accumulation. AZT treatment of C57BL/6J female mice (400 mg/day/kg body weight, i.p.) for 10 consecutive days significantly increased hepatic triglyceride levels and inflammation. Markers of oxidative stress such as protein oxidation, nitration, glycation and lipid peroxidation were significantly higher in the AZT-treated mice compared to vehicle controls. Further, the levels of ER stress marker proteins like GRP78, p-PERK, and p-eIF2α were significantly elevated in AZT-treated mice. The level of nuclear SREBP-1c, a transcription factor involved in fat synthesis, was increased while significantly decreased protein levels of phospho-acetyl-CoA carboxylase, phospho-AMP kinase and PPARα as well as inactivation of 3-keto-acyl-CoA thiolase in the mitochondrial fatty acid β-oxidation pathway were observed in AZT-exposed mice compared to those in control animals. Collectively, these data suggest that elevated oxidative and ER stress plays a key role, at least partially, in lipid accumulation, inflammation and hepatotoxicity in AZT-treated mice.


[1]  Hogg RS, Yip B, Kully C, Craib KJ, O’Shaughnessy MV, et al. (1999) Improved survival among HIV-infected patients after initiation of triple-drug antiretroviral regimens. CMAJ 160: 659–665.
[2]  Palella FJ Jr, Delaney KM, Moorman AC, Loveless MO, Fuhrer J, et al. (1998) Declining morbidity and mortality among patients with advanced human immunodeficiency virus infection. HIV Outpatient Study Investigators. N Engl J Med 338: 853–860.
[3]  Palella FJ Jr, Baker RK, Moorman AC, Chmiel JS, Wood KC, et al. (2006) Mortality in the highly active antiretroviral therapy era: changing causes of death and disease in the HIV outpatient study. J Acquir Immune Defic Syndr 43: 27–34.
[4]  Hansen L, Parker I, Monet Roberts L, Sutliff RL, Platt MO, et al. (2013) Azidothymidine (AZT) leads to arterial stiffening and intima-media thickening in mice. J Biomech 46: 1540–1547.
[5]  Georges B, Galland S, Rigault C, Le Borgne F, Demarquoy J (2003) Beneficial effects of L-carnitine in myoblastic C2C12 cells. Interaction with zidovudine. Biochem Pharmacol 65: 1483–1488.
[6]  Carr A, Miller J, Law M, Cooper DA (2000) A syndrome of lipoatrophy, lactic acidaemia and liver dysfunction associated with HIV nucleoside analogue therapy: contribution to protease inhibitor-related lipodystrophy syndrome. AIDS 14: F25–32.
[7]  Sutinen J, Hakkinen AM, Westerbacka J, Seppala-Lindroos A, Vehkavaara S, et al. (2002) Increased fat accumulation in the liver in HIV-infected patients with antiretroviral therapy-associated lipodystrophy. AIDS 16: 2183–2193.
[8]  Sulkowski MS, Thomas DL, Chaisson RE, Moore RD (2000) Hepatotoxicity associated with antiretroviral therapy in adults infected with human immunodeficiency virus and the role of hepatitis C or B virus infection. JAMA 283: 74–80.
[9]  Puoti M, Nasta P, Gatti F, Matti A, Prestini K, et al. (2009) HIV-related liver disease: ARV drugs, coinfection, and other risk factors. J Int Assoc Physicians AIDS Care (Chic) 8: 30–42.
[10]  Chariot P, Drogou I, de Lacroix-Szmania I, Eliezer-Vanerot MC, Chazaud B, et al. (1999) Zidovudine-induced mitochondrial disorder with massive liver steatosis, myopathy, lactic acidosis, and mitochondrial DNA depletion. J Hepatol 30: 156–160.
[11]  Freiman JP, Helfert KE, Hamrell MR, Stein DS (1993) Hepatomegaly with severe steatosis in HIV-seropositive patients. AIDS 7: 379–385.
[12]  Acosta BS, Grimsley EW (1999) Zidovudine-associated type B lactic acidosis and hepatic steatosis in an HIV-infected patient. South Med J 92: 421–423.
[13]  Corcuera T, Alonso MJ, Picazo A, Gomez F, Roldan M, et al. (1996) Hepatic Morphological alterations induced by zidovudine (ZDV) in an experimental model. Pathol Res Pract 192: 182–187.
[14]  Shah I (2006) Adverse effects of antiretroviral therapy in HIV-1 infected children. J Trop Pediatr 52: 244–248.
[15]  de la Asuncion JG, del Olmo ML, Sastre J, Millan A, Pellin A, et al. (1998) AZT treatment induces molecular and ultrastructural oxidative damage to muscle mitochondria. Prevention by antioxidant vitamins. J Clin Invest 102: 4–9.
[16]  de la Asuncion JG, del Olmo ML, Sastre J, Pallardo FV, Vina J (1999) Zidovudine (AZT) causes an oxidation of mitochondrial DNA in mouse liver. Hepatology 29: 985–987.
[17]  Szabados E, Fischer GM, Toth K, Csete B, Nemeti B, et al. (1999) Role of reactive oxygen species and poly-ADP-ribose polymerase in the development of AZT-induced cardiomyopathy in rat. Free Radic Biol Med 26: 309–317.
[18]  Manda KR, Banerjee A, Banks WA, Ercal N (2011) Highly active antiretroviral therapy drug combination induces oxidative stress and mitochondrial dysfunction in immortalized human blood-brain barrier endothelial cells. Free Radic Biol Med 50: 801–810.
[19]  Fromenty B, Pessayre D (1995) Inhibition of mitochondrial beta-oxidation as a mechanism of hepatotoxicity. Pharmacol Ther 67: 101–154.
[20]  Brinkman K, ter Hofstede HJ, Burger DM, Smeitink JA, Koopmans PP (1998) Adverse effects of reverse transcriptase inhibitors: mitochondrial toxicity as common pathway. AIDS 12: 1735–1744.
[21]  Abdelmegeed MA, Banerjee A, Yoo SH, Jang S, Gonzalez FJ, et al. (2012) Critical role of cytochrome P450 2E1 (CYP2E1) in the development of high fat-induced non-alcoholic steatohepatitis. J Hepatol 57: 860–866.
[22]  Moon KH, Hood BL, Kim BJ, Hardwick JP, Conrads TP, et al. (2006) Inactivation of oxidized and S-nitrosylated mitochondrial proteins in alcoholic fatty liver of rats. Hepatology 44: 1218–1230.
[23]  Abdelmegeed MA, Moon KH, Hardwick JP, Gonzalez FJ, Song BJ (2009) Role of peroxisome proliferator-activated receptor-alpha in fasting-mediated oxidative stress. Free Radic Biol Med 47: 767–778.
[24]  Kim BJ, Hood BL, Aragon RA, Hardwick JP, Conrads TP, et al. (2006) Increased oxidation and degradation of cytosolic proteins in alcohol-exposed mouse liver and hepatoma cells. Proteomics 6: 1250–1260.
[25]  Pagliassotti MJ (2012) Endoplasmic reticulum stress in nonalcoholic fatty liver disease. Annu Rev Nutr 32: 17–33.
[26]  Fu S, Watkins SM, Hotamisligil GS (2012) The role of endoplasmic reticulum in hepatic lipid homeostasis and stress signaling. Cell Metab 15: 623–634.
[27]  Desai VG, Lee T, Moland CL, Branham WS, Mittelstaedt RA, et al. (2012) Evaluation of Hepatic Mitochondria and Hematological Parameters in Zidovudine-Treated B6C3F(1) Mice. AIDS Res Treat 2012: 317695.
[28]  Omar RF, Gourde P, Desormeaux A, Tremblay M, Beauchamp D, et al. (1996) In vivo toxicity of foscarnet and zidovudine given alone or in combination. Toxicol Appl Pharmacol 139: 324–332.
[29]  Moon KH, Hood BL, Mukhopadhyay P, Rajesh M, Abdelmegeed MA, et al. (2008) Oxidative inactivation of key mitochondrial proteins leads to dysfunction and injury in hepatic ischemia reperfusion. Gastroenterology 135: 1344–1357.
[30]  Chen Q, Vazquez EJ, Moghaddas S, Hoppel CL, Lesnefsky EJ (2003) Production of reactive oxygen species by mitochondria: central role of complex III. J Biol Chem 278: 36027–36031.
[31]  Moon KH, Upreti VV, Yu LR, Lee IJ, Ye X, et al. (2008) Mechanism of 3,4-methylenedioxymethamphetamine (MDMA, ecstasy)-mediated mitochondrial dysfunction in rat liver. Proteomics 8: 3906–3918.
[32]  Abdelmegeed MA, Jang S, Banerjee A, Hardwick JP, Song BJ (2013) Robust protein nitration contributes to acetaminophen-induced mitochondrial dysfunction and acute liver injury. Free Radic Biol Med Epub ahead of print.
[33]  Kimzey AL, Weitz KK, Guengerich FP, Zangar RC (2003) Hydroperoxy-10,12-octadecadienoic acid stimulates cytochrome P450 3A protein aggregation by a mechanism that is inhibited by substrate. Biochemistry 42: 12691–12699.
[34]  Buechler C, Weiss TS (2011) Does hepatic steatosis affect drug metabolizing enzymes in the liver? Curr Drug Metab 12: 24–34.
[35]  Robertson G, Leclercq I, Farrell GC (2001) Nonalcoholic steatosis and steatohepatitis. II. Cytochrome P-450 enzymes and oxidative stress. Am J Physiol Gastrointest Liver Physiol 281: G1135–1139.
[36]  Lapadula G, Izzo I, Costarelli S, Cologni G, Bercich L, et al. (2007) Dideoxynucleoside HIV reverse transcriptase inhibitors and drug-related hepatotoxicity: a case report. J Med Case Rep 1: 19.
[37]  Sulkowski MS, Mehta SH, Chaisson RE, Thomas DL, Moore RD (2004) Hepatotoxicity associated with protease inhibitor-based antiretroviral regimens with or without concurrent ritonavir. AIDS 18: 2277–2284.
[38]  Miller TL, Evans SJ, Orav EJ, Morris V, McIntosh K, et al. (1993) Growth and body composition in children infected with the human immunodeficiency virus-1. Am J Clin Nutr 57: 588–592.
[39]  Calvaruso V, Craxi A (2009) Implication of normal liver enzymes in liver disease. J Viral Hepat 16: 529–536.
[40]  Wu D, Wang X, Zhou R, Yang L, Cederbaum AI (2012) Alcohol steatosis and cytotoxicity: the role of cytochrome P4502E1 and autophagy. Free Radic Biol Med 53: 1346–1357.
[41]  Hardwick JP, Osei-Hyiaman D, Wiland H, Abdelmegeed MA, Song BJ (2009) PPAR/RXR Regulation of Fatty Acid Metabolism and Fatty Acid omega-Hydroxylase (CYP4) Isozymes: Implications for Prevention of Lipotoxicity in Fatty Liver Disease. PPAR Res 2009: 952734.
[42]  Leclercq IA, Farrell GC, Field J, Bell DR, Gonzalez FJ, et al. (2000) CYP2E1 and CYP4A as microsomal catalysts of lipid peroxides in murine nonalcoholic steatohepatitis. Journal of Clinical Investigation 105: 1067–1075.
[43]  Wu J, Zhao MY, Zheng H, Zhang H, Jiang Y (2010) Pentoxifylline alleviates high-fat diet-induced non-alcoholic steatohepatitis and early atherosclerosis in rats by inhibiting AGE and RAGE expression. Acta Pharmacol Sin 31: 1367–1375.
[44]  Guimaraes EL, Empsen C, Geerts A, van Grunsven LA (2010) Advanced glycation end products induce production of reactive oxygen species via the activation of NADPH oxidase in murine hepatic stellate cells. J Hepatol 52: 389–397.
[45]  Gao RY, Mukhopadhyay P, Mohanraj R, Wang H, Horvath B, et al. (2011) Resveratrol attenuates azidothymidine-induced cardiotoxicity by decreasing mitochondrial reactive oxygen species generation in human cardiomyocytes. Mol Med Rep 4: 151–155.
[46]  Kaplowitz N, Than TA, Shinohara M, Ji C (2007) Endoplasmic reticulum stress and liver injury. Semin Liver Dis 27: 367–377.
[47]  Gentile CL, Frye MA, Pagliassotti MJ (2011) Fatty acids and the endoplasmic reticulum in nonalcoholic fatty liver disease. Biofactors 37: 8–16.
[48]  Harding HP, Zhang Y, Ron D (1999) Protein translation and folding are coupled by an endoplasmic-reticulum-resident kinase. Nature 397: 271–274.
[49]  Shi Y, Taylor SI, Tan SL, Sonenberg N (2003) When translation meets metabolism: multiple links to diabetes. Endocr Rev 24: 91–101.
[50]  Pahl HL, Baeuerle PA (1995) A novel signal transduction pathway from the endoplasmic reticulum to the nucleus is mediated by transcription factor NF-kappa B. EMBO J. 14: 2580–2588.
[51]  Wu S, Tan M, Hu Y, Wang JL, Scheuner D, et al. (2004) Ultraviolet light activates NFkappaB through translational inhibition of IkappaBalpha synthesis. J Biol Chem 279: 34898–34902.
[52]  Shimano H, Horton JD, Hammer RE, Shimomura I, Brown MS, et al. (1996) Overproduction of cholesterol and fatty acids causes massive liver enlargement in transgenic mice expressing truncated SREBP-1a. J Clin Invest 98: 1575–1584.
[53]  Kammoun HL, Chabanon H, Hainault I, Luquet S, Magnan C, et al. (2009) GRP78 expression inhibits insulin and ER stress-induced SREBP-1c activation and reduces hepatic steatosis in mice. J Clin Invest 119: 1201–1215.
[54]  Zhou H, Pandak WM Jr, Lyall V, Natarajan R, Hylemon PB (2005) HIV protease inhibitors activate the unfolded protein response in macrophages: implication for atherosclerosis and cardiovascular disease. Mol Pharmacol 68: 690–700.
[55]  You M, Fischer M, Deeg MA, Crabb DW (2002) Ethanol induces fatty acid synthesis pathways by activation of sterol regulatory element-binding protein (SREBP). J Biol Chem 277: 29342–29347.
[56]  Long YC, Zierath JR (2006) AMP-activated protein kinase signaling in metabolic regulation. J Clin Invest 116: 1776–1783.
[57]  Hardie DG, Carling D, Carlson M (1998) The AMP-activated/SNF1 protein kinase subfamily: metabolic sensors of the eukaryotic cell? Annu Rev Biochem 67: 821–855.
[58]  You M, Crabb DW (2004) Recent advances in alcoholic liver disease II. Minireview: molecular mechanisms of alcoholic fatty liver. Am J Physiol Gastrointest Liver Physiol 287: G1–6.
[59]  Garcia-Villafranca J, Guillen A, Castro J (2008) Ethanol consumption impairs regulation of fatty acid metabolism by decreasing the activity of AMP-activated protein kinase in rat liver. Biochimie 90: 460–466.


comments powered by Disqus