Impairment of liver mitochondrial β-oxidation is an important mechanism of drug-induced liver injury. Four inhibitors of fatty acid oxidation were compared in short-term rat in vivo studies in which the rats were administered one or four doses. The hepatocellular vacuolation represented ultrastructural mitochondrial changes. Urine nuclear magnetic resonance (NMR) spectroscopy revealed that both FOX988 and SDZ51-641 induced a persistent dicarboxylic aciduria, suggesting an inhibition of mitochondrial β-oxidation and incomplete fatty acid metabolism. Etomoxir caused minimal mitochondrial ultrastructural changes and induced only transient dicarboxylic aciduria. CPI975 served as a negative control, in that there were no significant perturbations to the mitochondrial ultrastructural morphology or in the urine NMR composition; however, compound exposure was confirmed by the up-regulation of liver gene expression compared to vehicle control. The liver gene expression changes that were altered by the compounds were indicative of mitochondria, general and oxidative stress, and peroxisomal enzymes involved in β-oxidation, suggestive of a compensatory response to the inhibition in the mitochondria. In addition, both FOX988 and SDZ51-641 up-regulated ribosomal genes associated with apoptosis, as well as p53 pathways linked with apoptosis. In summary, metabonomics and liver gene expression provided mechanistic information on mitochondrial dysfunction and impaired fatty acid oxidation to further define the clinical pathology and histopathology findings of hepatotoxicity.
Aldridge, J.E., Gibbon, J.A., Flaherty, M.M., et al. (2003). Heterogeneity of toxicant response: sources of human variability. Toxicol Sci76, 3-20.
2.
Anderson, R. ( 1998). Carnitine palmitoyltransferase: A viable target for the treatment of NIDDM? Current Pharmaceutical Design4, 1-15.
3.
Anderson, R., Balestr, M., Bell, P., Deems, R.O., Fillers, W.S., Foley, J.E., Fraser, J.D., Mann, W.R., Rudin, M., and Villhauer E.B. ( 1995). Antidiabetic agents: A new class of reversible carnitine palmitoyltransferase I inhibitors. J Med Chem38, 3448-3450.
4.
Beckwith-Hall, B.M., Nicholson, J.K., Nicholls, A.W., Foxall, P.J., Lindon, J.C., Connor, S.C., Abdi, M., Connelly, J., and Holmes, E. ( 1998). Nuclear magnetic resonance spectroscopic and principal components analysis investigations into biochemical affects of three model hepatotoxins. Chem Res Toxicol11, 260-72.
5.
Beckwith-Hall, B.M., Holmes, E., Lindon, J.C., Gounarides, J., Vickers, A., Shapiro, M., and Nicholson, J.K. ( 2002). NMR-based metabonomic studies on the biochemical effects of commonly used drug carrier vehicles in the rat. Chem Res Toxicol15, 1136-41.
6.
Brandt, J., Djouadi, F., and Kelly, D.P. ( 1998). Fatty acids activate transcription of the muscle carnitine palmitoyltransferase I gene in cardiac myocytes via the peroxisome proliferator-activated receptor α. J Biol Chem273, 23786-92.
7.
Burczynski, M.E., McMillian, M., Ciervo, J., Li, L., Parker, J.B., Dunn, R.T. 2nd, Hicken, S., Farr, S., and Johnson, M.D. ( 2000) Toxicogenomicsbased discrimination of toxic mechanism in HepG2 human hepatoma cells. Toxicol Sci58, 399-415.
8.
Cabrero, A., Merlos, M., Laguna, J.C., and Carrera, M.V. ( 2003). Down-regulation of acyl-CoA oxidase gene expression and increased NF-kB activity in etomoxir-induced cardiac hypertrophy. J Lipid Res44, 388-98.
9.
Chan, K., Truong, D., Shangari, N., and O’Brien, P.J. (2005). Drug-induced mitochondrial toxicity. Expert Opin Drug Metab Toxicol1, 655-69.
10.
Chen, F.W., and Ioannou, Y.A. ( 1999). Ribosomal proteins in cell proliferation and apoptosis . Int Rev Immunol18, 429-48.
11.
Deems, R.O., Anderson, R.C., and Foley, J.E. ( 1998). Hypoglycemic effects of a novel fatty acid oxidation inhibitor in rats and monkeys. Am J Physiol274, R524-28.
12.
Faergeman, N.J., and Knudsen, J. ( 1997). Role of long chain fatty acyl-CoA esters in the regulation of metabolism and in cell signaling. Biochem J323, 1-12.
13.
Fernandez-Checa, J., Kaplowitz, N., Garcia-Ruiz, C., and Colell, A. ( 1998) Mitochondrial Glutathione: Importance and Transport . Sem Liver Dis18, 389-401.
14.
Foley, J. ( 1992). Rationale and application of fatty acid oxidation inhibitors in treatment of diabetes mellitus. Diabetes Care15, 773-84.
15.
Foley, J., Anderson, R., Bell, P., Burkey, B.F., Deems, R.O., De Souza, C., and Dunning, B.E. ( 1997). Pharmacological strategies for reduction of lipid availability . Ann N Y Acad Sci827, 231-45.
16.
Forman, B.M., Chen, J., and Evans, R.M. ( 1997). Hypolipidemic drugs, polyunsaturated fatty acids, and eicosanoids are ligands for peroxisome proliferator-activated receptors α and δ. Proc Natl Acad Sci USA94, 4312-17.
17.
Fromenty, B., and Pessayre, D. ( 1995). Inhibition of mitochondrial beta-oxidation as a mechanism of hepatotoxicity. Pharmacol Ther67, 101-54.
18.
Greaves, P., Williams, A., and Eve, M. ( 2004). First dose of potential new medicines to humans: How animals help. Nat Rev Drug Disc3, 226-36.
Gunawan, B.K., and Kaplowitz, N. ( 2007). Mechanisms of drug-induced liver disease. Clin Liver Dis11, 459-75.
21.
Heilbronn, L.K., Gan, S.K., Turner, N., Campbell, L.V., and Chisholm, D.J. ( 2007). Markers of mitochondrial biogenesis and metabolism are lower in overweight and obese insulin-resistant subjects. J Clin Endocrinol Metab92, 1467-73.
22.
Ho, J.K., Duclos, R.I. Jr., and Hamilton, J.A. ( 2002). Interaction of acylcarnitines with model membranes: a (13)C-NMR study. J Lipid Res43, 1429-39.
23.
Horn, C.C., Ji, H., and Friedman, M.I. ( 2004). Etomoxir, a fatty acid oxidation inhibitor, increases food intake and reduces hepatic energy status in rats. Physiol Behav81, 157-62.
24.
Kahler, A., Zimmermann, M., and Langhans, W. ( 1999). Suppression of hepatic fatty acid oxidation and food intake in men. Nutrition15, 819-28.
25.
Klaassen, C.D., Liu, J., and Choudhuri, S. ( 1999). Metallothionein: an intracellular protein to protect against cadmium toxicity. Annu Rev Pharmacol Toxicol39, 267-94.
26.
Laskin, J.D., Heck, D.E., and Laskin, D.L. ( 2002). The ribotoxic stress response as a potential mechanism for MAP kinase activation in xenobiotic toxicity. Toxicol Sci69, 289-91.
27.
Lee, L., Horowitz, J., and Frenneaux, M. ( 2004). Metabolic manipulation in ischaemic heart disease, a novel approach to treatment. Eur Heart J25, 634-41.
28.
Lock, E., Mitchell, A.M., and Elcombe, C.R. ( 1989). Biochemical mechanisms of induction of hepatic peroxisome proliferation. Annu Rev Pharmacol Toxicol29,145-63.
29.
Lopaschuk, G.D., and Stanley, W.C. ( 1997). Manipulation of energy metabolism in the heart . Sci Med4, 42-51.
30.
Maher, J. ( 1999) Cytokines: Overview. Sem Liver Dis19, 109-15.
31.
Marazzi, G., Volterrani, M., and Rosano, G.M.C.Metabolic agents in the management of diabetic coronary patients: a new era . Int J Cardiology, in press.
32.
Mazumder, B., Sampath, P., Seshadri, V., Maitra, R.K., DiCorleto, P.E., and Fox, P.L. ( 2003). Regulated release of L13a from the 60S ribosomal subunit as a mechanism of transcript-specific translational control. Cell115, 187-98.
33.
McKenzie, R., Fried, M.W., Sallie, R., Conjeevaram, H., Di Bisceglie, A.M., Park, Y., Savarese, B., Kleiner, D., Tsokos, M., Luciano, C., Pruett, T., Stotka, J.L., Straus, S.E., and Hoofnagle, J.H. ( 1995). Hepatic failure and lactic acidosis due to fialuridine (FIAU), an investigational nucleoside analogue for chronic hepatitis B. N Engl J Med333, 1099-105.
Mortensen, P.B. ( 1990). Mechanisms of dicarboxylic aciduria and dicarboxylic acid metabolism. In Fatty Acid Oxidation: Clinical, Biochemical and Molecular Aspects, pp 249-64, Alan R. Liss, Inc., New York.
36.
Murray, A.J., Panagia, M., Hauton, D., Gibbons, G.F., and Clarke, K. ( 2005). Plasma free fatty acids and peroxisome proliferator-activated receptor α in the control of myocardial uncoupling protein levels. Diabetes54, 3496-502.
37.
Nada, M.A., Chace, D.H., Sprecher, H., and Roe, C.R. ( 1995). Investigation of beta-oxidation intermediates in normal and MCAD deficient human fibroblasts using tandem mass spectrometry . Biochem Molec Med54, 59-66.
38.
Olson, H., Betton, G., Robinson, D., Thomas, K., Munro, A., Kolaja, G., Lilly, P., Sanders, J., Sipes, G., Bracken, W., Dorato, M., Van Deun, K., Smith, P., Berger, B., and Heller, A. ( 2000). Concordance of the toxicity of pharmaceuticals in humans and animals. Regul Toxicol Pharmacol32, 56-67.
39.
Pessayree, D., Mansouri, A., Haouzi, D., and Fromenty, B. ( 1999). Hepatotoxicity due to mitochondrial dysfunction . Cell Biol Toxicol15, 367-73.
40.
Reddy, J.K. ( 2001). Nonalcoholic steatosis and steatohepatitis III. Peroxisomal β-oxidation, PPARα, and steatohepatitis. Am J Physiol Gastrointest Liver Physiol281, G1333-39.
41.
Rupp, H., and Jacob, R. ( 1992). Metabolically modulated growth and phenotype of the rat heart. Eur Heart J13, 56-61.
42.
Schmeichel, K.L., and Bissell, M.J. ( 2003). Modeling tissue-specific signaling and organ function in three dimensions. J Cell Sci116, 2377-88.
43.
Schrauwen, P., Hinderling, V., Hesselink, M.K.C., Schaart, G., Kornips, E., Saris, W.H.M., Westerterp-Plantenga, M., and Langhans, W. ( 2002). Etomoxir-induced increase in UCP3 supports a role of uncoupling protein 3 as a mitochondrial fatty acid anion exporter. FASEB J16, 1688-90.
44.
Schrauwen, P., and Hesselink, M.K.C. (2004). Oxidative capacity, lipotoxicity, and mitochondrial damage in type 2 diabetes. Diabetes53, 1412-17.
45.
Schrauwen, P., Hoeks, J., and Hesselink, M.K. ( 2006). Putative function and physiological relevance of the mitochondrial uncoupling protein-3: involvement in fatty acid metabolism ? Prog Lipid Res45, 17-41.
46.
Schrauwen, P. ( 2008). Editorial: of the fit and the fat: mitochondrial abnormalities and type 2 diabetes mellitus. J Clin Endocrin Metab92, 1229-31.
47.
Szewczyk, A., and Wojtczak, L. ( 2002). Mitochondria as a pharmacological target. Pharmacol Rev54, 101-27.
48.
Van de Water, B., de Graauw, M., Le Devedec, S., and Alderliesten, M. (2006). Cellular stress responses and molecular mechanisms of nephrotoxicity. Toxicol Lett162, 83-93.
49.
Vickers, A. E. M, and Fisher, R.L. ( 2005). Precision-cut organ slices to investigate target organ injury. Exp Opin Drug Metab Toxicol1, 687-99.
50.
Vickers, A.E.M., Bentley, P., and Fisher, R.L. ( 2006). Consequences of mitochondrial injury induced by pharmaceutical fatty acid oxidation inhibitors is characterized in human and rat liver slices . Toxicol In Vitro20, 1173-82.
51.
Weis, B.C., Cowan, A.T., Brown, N., Foster, D.W., and McGarry, J.D. ( 1994). Use of a selective inhibitor of liver carnitine palmitoyltransferase I (CPT I) allows quantification of its contribution to total CPT I activity in rat heart. J Biol Chem269, 26443-48.
52.
Wille, A., Gerber, A., Heimburg, A., Reisenauer, A., Peters, C., Saftig, P., Reinheckel, T., Welte, T., and Buhling, F. ( 2004). Cathepsin L is involved in cathepsin D processing and regulation of apoptosis in A549 human lung epithelial cells. Biol Chem385, 665-70.
53.
Wilson, M., and Easterbrook-Smith, S.B. (2000). Clusterin is a secreted mammalian chaperone. Trends Biochem Sci25, 95-98.
54.
Wiltshire, C., Black, S., and Brown, A.J. ( 1999). Over-expression of Candida albicans mitochondrial ribosomal protein S9 (MrpS9p) disturbs mitochondrial function in Saccharomyces cerevisiae . Yeast15, 139-43.
55.
Yotsumoto, T., Naitoh, T., Kitahara, M., and Tzuruzoe, N. ( 2000). Effects of carnitine palmitoyltransferase I inhibitors on hepatic hypertrophy. Eur J Pharmacol398, 297-302.
