Potential urinary and plasma biomarkers of peroxisome proliferation in the rat: identification of N-methylnicotinamide and N-methyl-4-pyridone-3-carboxamide by 1H nuclear magnetic resonance and high performance liquid chromatography

This study identified two potential novel biomarkers of peroxisome proliferation in the rat. Three peroxisome proliferator-activated receptor (PPAR) ligands, chosen for their high selectivity towards the PPARα, -δ and -γ subtypes, were given to rats twice daily for 7 days at doses known to cause a pharmacological effect or peroxisome proliferation. Fenofibrate was used as a positive control. Daily treatment with the PPARα and -δ agonists produced peroxisome proliferation and liver hypertrophy. 1H nuclear magnetic resonance spectroscopy and multivariate statistical data analysis of urinary spectra from animals given the PPARα and -δ agonists identified two new potential biomarkers of peroxisome proliferation - N-methylnicotinamide (NMN) and N-methyl-4-pyridone-3-carboxamide (4PY) - both endproducts of the tryptophan-nicotinamide adenine dinucleotide (NAD+) pathway. After 7 days, excretion of NMN and 4PY increased 24- and three-fold, respectively, following high doses of fenofibrate. The correlation between total NMN excretion over 7 days and the peroxisome count was r=0.87 (r2=0.76). Plasma NMN, measured using a sensitive high performance liquid chromatography method, was increased up to 61-fold after 7 days' treatment with high doses of fenofibrate. Hepatic gene expression of aminocarboxymuconate-semialdehyde decarboxylase (EC 4.1.1.45) was downregulated following treatment with the PPARα and -δ agonists. The decrease was up to 11-fold compared with controls in the groups treated with high doses of fenofibrate. This supports the link between increased NMN and 4PY excretion and regulation of the tryptophan-NAD+ pathway in the liver. In conclusion, NMN, and possibly other metabolites in the pathway, are potential non-invasive surrogate biomarkers of peroxisome proliferation in the rat.

[1]  Y. Nishizuka,et al.  STUDIES ON THE BIOSYNTHESIS OF NICOTINAMIDE ADENINE DINUCLEOTIDE. II. A ROLE OF PICOLINIC CARBOXYLASE IN THE BIOSYNTHESIS OF NICOTINAMIDE ADENINE DINUCLEOTIDE FROM TRYPTOPHAN IN MAMMALS. , 1965, The Journal of biological chemistry.

[2]  P Grasso,et al.  Hepatic peroxisome proliferation in rodents and its significance for humans. , 1993, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[3]  S. Yuyama,et al.  Isolation and identification of N1-methylnicotinic acid (trigonelline) from rat urine. , 1985, Journal of nutritional science and vitaminology.

[4]  J. Styles,et al.  Mitosis and histopathology in rat liver during methylclofenapate-induced hyperplasia. , 1990, Cancer Letters.

[5]  S Kettle,et al.  Mechanistically-based Human Hazard Assessment of Peroxisome Proliferator-induced Hepatocarcinogenesis , 1994, Human & experimental toxicology.

[6]  N. Lalwani,et al.  Induction of fatty acid beta-oxidation and peroxisome proliferation in the liver of rhesus monkeys by DL-040, a new hypolipidemic agent. , 1985, Biochemical pharmacology.

[7]  K. Shibata,et al.  Niacin catabolism in rodents. , 1990, Journal of nutritional science and vitaminology.

[8]  C. Elcombe,et al.  Peroxisome Proliferators: Species Differences in Response of Primary Hepatocyte Cultures , 1996, Annals of the New York Academy of Sciences.

[9]  E. Ros,et al.  Fibrate treatment does not modify the expression of acyl coenzyme A oxidase in human liver , 2002, Clinical pharmacology and therapeutics.

[10]  S. Wold,et al.  Orthogonal signal correction of near-infrared spectra , 1998 .

[11]  B. Lake.  Peroxisome proliferation: current mechanisms relating to nongenotoxic carcinogenesis. , 1995, Toxicology letters.

[12]  C. Bernofsky.  New synthesis of the 4- and 6-pyridones of 1-methylnicotinamide and 1-methylnicotinic acid (trigonelline). , 1979, Analytical biochemistry.

[13]  K. Beier.  Light microscopic morphometric analysis of peroxisomes by automatic image analysis: advantages of immunostaining over the alkaline DAB method. , 1992, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[14]  Y. Egashira,et al.  Change of tryptophan-niacin metabolism in D-galactosamine-induced liver injury in rat. , 1997, Journal of nutritional science and vitaminology.

[15]  C Kemmer,et al.  Relationship between morphological changes and lipid-lowering action of p-chlorphenoxyisobutyric acid (CPIB) on hepatic mitochondria and peroxisomes in man. , 1983, Atherosclerosis.

[16]  K. Shibata,et al.  Influence of Adenine-induced Renal Failure on Tryptophan-niacin Metabolism in Rats , 2001, Bioscience, biotechnology, and biochemistry.

[17]  M. Hollinshead,et al.  Immunocytochemical analysis of soluble epoxide hydrolase and catalase in mouse and rat hepatocytes demonstrates a peroxisomal localization before and after clofibrate treatment. , 1988, European journal of cell biology.

[18]  Y. Egashira,et al.  Expression of rat hepatic 2-amino-3-carboxymuconate-6-semialdehyde decarboxylase is affected by a high protein diet and by streptozotocin-induced diabetes. , 2002, The Journal of nutrition.

[19]  R. Gholson,et al.  Isolation and identification of two new nicotinamide metabolites. , 1969, The Journal of biological chemistry.

[20]  R. Cuomo,et al.  Nicotinamide methylation and hepatic energy reserve: a study by liver perfusion in vitro. , 1995, Journal of hepatology.

[21]  J. Nicholson,et al.  Hepatotoxin-induced hypertaurinuria: a proton NMR study , 2005, Archives of Toxicology.

[22]  M. Shin,et al.  NAD+ biosynthesis and metabolic fluxes of tryptophan in hepatocytes isolated from rats fed a clofibrate-containing diet. , 1996, Biochemical pharmacology.

[23]  B. Ledwith,et al.  Peroxisome proliferators activate growth regulatory pathways largely via peroxisome proliferator-activated receptor alpha-independent mechanisms. , 2002, Cellular signalling.

[24]  B. Lake,et al.  Peroxisome proliferation and nongenotoxic carcinogenesis: commentary on a symposium. , 1991, Fundamental and applied toxicology : official journal of the Society of Toxicology.

[25]  M. Shin,et al.  Effects of peroxisome-proliferators on the TRP-NAD pathway. , 1999, Advances in experimental medicine and biology.

[26]  C. Dunnett.  A Multiple Comparison Procedure for Comparing Several Treatments with a Control , 1955 .

[27]  B. Lake,et al.  Peroxisome proliferation and nongenotoxic carcinogenesis: Commentary on a symposium , 1991 .

[28]  Y. Egashira,et al.  Suppression of Rat Hepatic α-Amino-β-carboxymuconate-ε-semialdehyde Decarboxylase (ACMSD) Activity by Linoleic Acid in Relation to Its Induction by Glucocorticoids and Dietary Protein , 1994 .

[29]  Andrew W Nicholls Jeremy K Nicholson John N Haseld.  A metabonomic approach to the investigation of drug-induced phospholipidosis: an NMR spectroscopy and pattern recognition study. , 2000, Biomarkers : biochemical indicators of exposure, response, and susceptibility to chemicals.

[30]  N. Iwamoto,et al.  Pyridine nucleotide levels in liver of rats fed clofibrate- or pyrazinamide-containing diets. , 1998, Biochemical pharmacology.

[31]  K. Shibata,et al.  Elucidation of the Toxic Mechanism of the Plasticizers, Phthalic Acid Esters, Putative Endocrine Disrupters: Effects of Dietary Di(2-ethylhexyl)phthalate on the Metabolism of Tryptophan to… , 2002, Bioscience, biotechnology, and biochemistry.

[32]  H. Wolf.  Studies on Tryptophan Metabolism in Man: The Effect of Hormones and Vitamin B6 on Urinary Excretion of Metabolites of the Kynurenine Pathway: Part 2 , 1974 .

[33]  H. Wolf.  The effect of hormones and vitamin B6 on urinary excretion of metabolites of the kynurenine pathway. , 1974, Scandinavian journal of clinical and laboratory investigation. Supplementum.

[34]  N. Lalwani,et al.  Induction of hepatic peroxisome proliferation in nonrodent species, including primates. , 1984, The American journal of pathology.

[35]  P. Novikoff,et al.  Catalase-negative peroxisomes: transient appearance in rat hepatocytes during liver regeneration after partial hepatectomy. , 1995, The American journal of pathology.

[36]  J. Biollaz,et al.  Validation of an HPLC method for the determination of urinary and plasma levels of N1-methylnicotinamide, an endogenous marker of renal cationic transport and plasma flow. , 2001, Journal of pharmaceutical and biomedical analysis.

[37]  F J Gonzalez,et al.  Evidence for peroxisome proliferator-activated receptor (PPAR)alpha-independent peroxisome proliferation: effects of PPARgamma/delta-specific agonists in PPARalpha-null mice. , 2000, Molecular pharmacology.

[38]  J. Lehmann,et al.  N-(2-Benzoylphenyl)-L-tyrosine PPARgamma agonists. 1. Discovery of a novel series of potent antihyperglycemic and antihyperlipidemic agents. , 1998, Journal of medicinal chemistry.

[39]  B. Lake.  Mechanisms of hepatocarcinogenicity of peroxisome-proliferating drugs and chemicals. , 1995, Annual review of pharmacology and toxicology.

[40]  P J Sadler,et al.  Quantitative high resolution 1H NMR urinalysis studies on the biochemical effects of cadmium in the rat. , 1989, Molecular pharmacology.

[41]  Y. Egashira,et al.  Effects of Various Dietary Fatty Acids on α-Amino-β-carboxymuconate-ε-semialdehyde Decarboxylase Activity in Rat Liver , 1992 .

[42]  Y. Egashira,et al.  Dietary linoleic acid alters alpha-amino-beta-carboxymuconate-epsilon-semialdehyde decarboxylase (ACMSD), a key enzyme of niacin synthesis from tryptophan, in the process of protein expression in rat liver. , 1998, Journal of nutritional science and vitaminology.

[43]  R. Cuomo,et al.  The metabolism of nicotinamide in human liver cirrhosis: a study on N-methylnicotinamide and 2-pyridone-5-carboxamide production , 2001, American Journal of Gastroenterology.

[44]  Brian C Sweatman,et al.  Optimisation of collection, storage and preparation of rat plasma for 1H NMR spectroscopic analysis in toxicology studies to determine inherent variation in biochemical profiles. , 2002, Journal of pharmaceutical and biomedical analysis.

[45]  J. Lindon,et al.  'Metabonomics': understanding the metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data. , 1999, Xenobiotica; the fate of foreign compounds in biological systems.

[46]  M. Shin,et al.  Effect of Thioridazine or Chlorpromazine on Increased Hepatic NAD+ Level in Rats Fed Clofibrate, a Hypolipidaemic Drug , 1998, The Journal of pharmacy and pharmacology.