Fatty liver or hepatic steatosis is a common health problem associated with abnormal liver function and increased susceptibility to ischemia/reperfusion injury. The objective of this study was to investigate the effect of the fatty acid synthase inhibitor cerulenin on hepatic function in steatotic ob/ob mice. Different dosages of cerulenin were administered intraperitoneally to ob/ob mice for 2 to 7 days. Body weight, serum AST/ALT, hepatic energy state, and gene expression patterns in ob/ob mice were examined. We found that cerulenin treatment markedly improved hepatic function in ob/ob mice. Serum AST/ALT levels were significantly decreased and hepatic ATP levels increased in treated obese mice compared to obese controls, accompanied by fat depletion in the hepatocyte. Expression of peroxisome proliferator-activated receptors α and γ and uncoupling protein 2 were suppressed with cerulenin treatment and paralleled changes in AST/ALT levels. Hepatic glutathione content were increased in some cases and apoptotic activity in the steatotic livers was minimally changed with cerulenin treatment. In conclusion, these results demonstrate that fatty acid synthase blockade constitutes a novel therapeutic strategy for altering hepatic steatosis at non-stressed states in obese livers.
References
[1]
Falck-Ytter Y, Younossi ZM, Marchesini G, McCullough AJ (2001) Clinical features and natural history of nonalcoholic steatosis syndromes. Semin Liver Dis 21: 17–26.
Farrell GC, Larter CZ (2006) Nonalcoholic fatty liver disease: from steatosis to cirrhosis. Hepatology 43: S99–S112.
[4]
Della Corte C, Alisi A, Saccari A, De Vito R, Vania A, et al. (2012) Nonalcoholic fatty liver in children and adolescents: an overview. J Adolesc Health 51: 305–312.
[5]
Nadig SN, Periyasamy B, Shafizadeh SF, Polito C, Fiorini RN, et al. (2004) Hepatocellular ultrastructure after ischemia/reperfusion injury in human orthotopic liver transplantation. J Gastrointest Surg 8: 695–700.
[6]
Malnick SD, Beergabel M, Knobler H (2003) Non-alcoholic fatty liver: a common manifestation of a metabolic disorder. QJM 96: 699–709.
[7]
Begriche K, Massart J, Robin MA, Bonnet F, Fromenty B (2013) Mitochondrial adaptations and dysfunctions in nonalcoholic fatty liver disease. Hepatology.
[8]
Basaranoglu M, Basaranoglu G, Senturk H (2013) From fatty liver to fibrosis: A tale of “second hit”. World J Gastroenterol 19: 1158–1165.
Browning JD, Horton JD (2004) Molecular mediators of hepatic steatosis and liver injury. J Clin Invest 114: 147–152.
[11]
Nardo B, Caraceni P, Pasini P, Domenicali M, Catena F, et al. (2001) Increased generation of reactive oxygen species in isolated rat fatty liver during postischemic reoxygenation. Transplantation 71: 1816–1820.
[12]
Chavin KD, Yang S, Lin HZ, Chatham J, Chacko VP, et al. (1999) Obesity induces expression of uncoupling protein-2 in hepatocytes and promotes liver ATP depletion. J Biol Chem 274: 5692–5700.
[13]
Nakatani T, Tsuboyama-Kasaoka N, Takahashi M, Miura S, Ezaki O (2002) Mechanism for peroxisome proliferator-activated receptor-alpha activator-induced up-regulation of UCP2 mRNA in rodent hepatocytes. J Biol Chem 277: 9562–9569.
[14]
Kimura K, Jung BD, Kanehira K, Irie Y, Canas X, et al. (1999) Induction of uncoupling protein (UCP) 2 in primary cultured hepatocytes. FEBS Lett 457: 75–79.
[15]
Van Der Lee KA, Willemsen PH, Van Der Vusse GJ, Van Bilsen M (2000) Effects of fatty acids on uncoupling protein-2 expression in the rat heart. FASEB J 14: 495–502.
[16]
Furukawa S, Fujita T, Shimabukuro M, Iwaki M, Yamada Y, et al. (2004) Increased oxidative stress in obesity and its impact on metabolic syndrome. J Clin Invest 114: 1752–1761.
[17]
Asselah T, Rubbia-Brandt L, Marcellin P, Negro F (2006) Steatosis in chronic hepatitis C: why does it really matter? Gut 55: 123–130.
Menendez JA, Vellon L, Mehmi I, Oza BP, Ropero S, et al. (2004) Inhibition of fatty acid synthase (FAS) suppresses HER2/neu (erbB-2) oncogene overexpression in cancer cells. Proc Natl Acad Sci U S A 101: 10715–10720.
[20]
Nomura S, Horiuchi T, Hata T, Omura S (1972) Inhibition of sterol and fatty acid biosyntheses by cerulenin in cell-free systems of yeast. J Antibiot (Tokyo) 25: 365–368.
[21]
D’Agnolo G, Rosenfeld IS, Awaya J, Omura S, Vagelos PR (1973) Inhibition of fatty acid synthesis by the antibiotic cerulenin. Specific inactivation of beta-ketoacyl-acyl carrier protein synthetase. Biochim Biophys Acta 326: 155–156.
[22]
Price AC, Choi KH, Heath RJ, Li Z, White SW, et al. (2001) Inhibition of beta-ketoacyl-acyl carrier protein synthases by thiolactomycin and cerulenin. Structure and mechanism. J Biol Chem 276: 6551–6559.
[23]
Ronnett GV, Kleman AM, Kim EK, Landree LE, Tu Y (2006) Fatty acid metabolism, the central nervous system, and feeding. Obesity (Silver Spring) 14 Suppl 5201S–207S.
[24]
Loftus TM, Jaworsky DE, Frehywot GL, Townsend CA, Ronnett GV, et al. (2000) Reduced food intake and body weight in mice treated with fatty acid synthase inhibitors. Science 288: 2379–2381.
[25]
Dridi S, Ververken C, Hillgartner FB, Arckens L, Van der Gucht E, et al. (2006) FAS inhibitor cerulenin reduces food intake and melanocortin receptor gene expression without modulating the other (an)orexigenic neuropeptides in chickens. Am J Physiol Regul Integr Comp Physiol 291: R138–147.
[26]
Chavin KD, Fiorini RN, Shafizadeh S, Cheng G, Wan C, et al. (2004) Fatty acid synthase blockade protects steatotic livers from warm ischemia reperfusion injury and transplantation. Am J Transplant 4: 1440–1447.
[27]
Makimura H, Mizuno TM, Yang XJ, Silverstein J, Beasley J, et al. (2001) Cerulenin mimics effects of leptin on metabolic rate, food intake, and body weight independent of the melanocortin system, but unlike leptin, cerulenin fails to block neuroendocrine effects of fasting. Diabetes 50: 733–739.
[28]
Cheng G, Polito CC, Haines JK, Shafizadeh SF, Fiorini RN, et al. (2003) Decrease of intracellular ATP content downregulated UCP2 expression in mouse hepatocytes. Biochem Biophys Res Commun 308: 573–580.
[29]
Marques AR, Espadinha C, Frias MJ, Roque L, Catarino AL, et al. (2004) Underexpression of peroxisome proliferator-activated receptor (PPAR)gamma in PAX8/PPARgamma-negative thyroid tumours. Br J Cancer 91: 732–738.
[30]
Folch J, Lees M, Sloane Stanley GH (1957) A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem 226: 497–509.
[31]
Baker MA, Cerniglia GJ, Zaman A (1990) Microtiter plate assay for the measurement of glutathione and glutathione disulfide in large numbers of biological samples. Anal Biochem 190: 360–365.
[32]
Cho WS, Cho M, Jeong J, Choi M, Cho HY, et al. (2009) Acute toxicity and pharmacokinetics of 13 nm-sized PEG-coated gold nanoparticles. Toxicol Appl Pharmacol 236: 16–24.
[33]
Evans ZP, Mandavilli BS, Ellett JD, Rodwell D, Fariss MW, et al. (2009) Vitamin E succinate enhances steatotic liver energy status and prevents oxidative damage following ischemia/reperfusion. Transplant Proc 41: 4094–4098.
[34]
Rashid A, Wu TC, Huang CC, Chen CH, Lin HZ, et al. (1999) Mitochondrial proteins that regulate apoptosis and necrosis are induced in mouse fatty liver. Hepatology 29: 1131–1138.
[35]
Cortez-Pinto H, Zhi Lin H, Qi Yang S, Odwin Da Costa S, Diehl AM (1999) Lipids up-regulate uncoupling protein 2 expression in rat hepatocytes. Gastroenterology 116: 1184–1193.
[36]
Memon RA, Tecott LH, Nonogaki K, Beigneux A, Moser AH, et al. (2000) Up-regulation of peroxisome proliferator-activated receptors (PPAR-alpha) and PPAR-gamma messenger ribonucleic acid expression in the liver in murine obesity: troglitazone induces expression of PPAR-gamma-responsive adipose tissue-specific genes in the liver of obese diabetic mice. Endocrinology 141: 4021–4031.
[37]
Rahimian R, Masih-Khan E, Lo M, van Breemen C, McManus BM, et al. (2001) Hepatic over-expression of peroxisome proliferator activated receptor gamma2 in the ob/ob mouse model of non-insulin dependent diabetes mellitus. Mol Cell Biochem 224: 29–37.
[38]
Pizer ES, Jackisch C, Wood FD, Pasternack GR, Davidson NE, et al. (1996) Inhibition of fatty acid synthesis induces programmed cell death in human breast cancer cells. Cancer Res 56: 2745–2747.
[39]
Thupari JN, Pinn ML, Kuhajda FP (2001) Fatty acid synthase inhibition in human breast cancer cells leads to malonyl-CoA-induced inhibition of fatty acid oxidation and cytotoxicity. Biochem Biophys Res Commun 285: 217–223.
[40]
Guan LL, Wang YF, Gong DZ, Yuan B, Wu Q, et al. (2012) [Establishment of the Chang liver cell line stably overexpressing human UCP2 gene and its effect on mitochondrial membrane potential and reactive oxygen species]. Zhonghua Gan Zang Bing Za Zhi 20: 131–135.
[41]
Armeni T, Ghiselli R, Balercia G, Goffi L, Jassem W, et al. (2000) Glutathione and ultrastructural changes in inflow occlusion of rat liver. J Surg Res 88: 207–214.
[42]
Tontonoz P, Hu E, Spiegelman BM (1994) Stimulation of adipogenesis in fibroblasts by PPAR gamma 2, a lipid-activated transcription factor. Cell 79: 1147–1156.
[43]
Francis GA, Fayard E, Picard F, Auwerx J (2003) Nuclear receptors and the control of metabolism. Annu Rev Physiol 65: 261–311.
[44]
Dongiovanni P, Valenti L (2013) Peroxisome proliferator-activated receptor genetic polymorphisms and nonalcoholic Fatty liver disease: any role in disease susceptibility? PPAR Res 2013: 452061.
[45]
Gres S, Canteiro S, Mercader J, Carpene C (2013) Oxidation of high doses of serotonin favors lipid accumulation in mouse and human fat cells. Mol Nutr Food Res.
[46]
de Las Heras N, Valero-Munoz M, Ballesteros S, Gomez-Hernandez A, Martin-Fernandez B, et al.. (2013) Factors involved in rosuvastatin induction of insulin sensitization in rats fed a high fat diet. Nutr Metab Cardiovasc Dis.
[47]
Suzuki S, Suzuki M, Sembon S, Fuchimoto D, Onishi A (2013) Characterization of actions of octanoate on porcine preadipocytes and adipocytes differentiated in vitro. Biochem Biophys Res Commun 432: 92–98.
[48]
Spiegelman BM, Puigserver P, Wu Z (2000) Regulation of adipogenesis and energy balance by PPARgamma and PGC-1. Int J Obes Relat Metab Disord 24 Suppl 4S8–10.
[49]
Tang QQ, Lane MD (2012) Adipogenesis: from stem cell to adipocyte. Annu Rev Biochem 81: 715–736.
Berger J, Moller DE (2002) The mechanisms of action of PPARs. Annu Rev Med 53: 409–435.
[52]
Medvedev AV, Snedden SK, Raimbault S, Ricquier D, Collins S (2001) Transcriptional regulation of the mouse uncoupling protein-2 gene. Double E-box motif is required for peroxisome proliferator-activated receptor-gamma-dependent activation. J Biol Chem 276: 10817–10823.
[53]
Viguerie-Bascands N, Saulnier-Blache JS, Dandine M, Dauzats M, Daviaud D, et al. (1999) Increase in uncoupling protein-2 mRNA expression by BRL49653 and bromopalmitate in human adipocytes. Biochem Biophys Res Commun 256: 138–141.
[54]
Shimokawa T, Kato M, Watanabe Y, Hirayama R, Kurosaki E, et al. (1998) In vivo effects of pioglitazone on uncoupling protein-2 and -3 mRNA levels in skeletal muscle of hyperglycemic KK mice. Biochem Biophys Res Commun 251: 374–378.
[55]
Boelsterli UA, Bedoucha M (2002) Toxicological consequences of altered peroxisome proliferator-activated receptor gamma (PPARgamma) expression in the liver: insights from models of obesity and type 2 diabetes. Biochem Pharmacol 63: 1–10.
[56]
Matsusue K, Haluzik M, Lambert G, Yim SH, Gavrilova O, et al. (2003) Liver-specific disruption of PPARgamma in leptin-deficient mice improves fatty liver but aggravates diabetic phenotypes. J Clin Invest 111: 737–747.
[57]
Fink BD, Reszka KJ, Herlein JA, Mathahs MM, Sivitz WI (2005) Respiratory uncoupling by UCP1 and UCP2 and superoxide generation in endothelial cell mitochondria. Am J Physiol Endocrinol Metab 288: E71–79.
[58]
Deepa PR, Vandhana S, Krishnakumar S (2013) Fatty Acid synthase inhibition induces differential expression of genes involved in apoptosis and cell proliferation in ocular cancer cells. Nutr Cancer 65: 311–316.
[59]
Tirado-Velez JM, Joumady I, Saez-Benito A, Cozar-Castellano I, Perdomo G (2012) Inhibition of fatty acid metabolism reduces human myeloma cells proliferation. PLoS One 7: e46484.
