Induction of an ATPase inhibitor protein by propylthiouracil and protection against paracetamol (acetaminophen) hepatotoxicity in the rat

1 The purpose of the present study was to test the following hypothesis: propylthiouracil (PTU) treatments of rats induces an increase in the concentration and activity of the mitochondrial ATPase (m‐ATPase) inhibitor protein (IF1). The PTU‐induced elevated baseline levels of this inhibitor protein inactivated m‐ATPase, and prevented hepatotoxicity by a toxic dose of acetaminophen (AAP) (paracetamol), by maintaining hepatic adenosine 5′‐triphosphate (ATP) levels. 2 Male Wistar rats were either gavaged with a toxic dose of AAP alone, or after pretreatment with PTU for periods of 3 and 12 days. 3 Twenty four hours after acetaminophen treatment alone, toxicity was manifested by: an approximately 10 fold increase in serum transaminase levels (serum glutamic oxaloacetic transaminase and serum glutamic pyruvic transaminase); depletion of hepatic reduced glutathione (GSH) and ATP levels; loss of inhibitor protein activity, and extensive pericentral necrosis of the hepatocytes. Propylthiouracil pretreatment for 12 days enhanced the concentration of the following metabolites in the liver: ATP (1.5 fold), ATPase inhibitor protein (IF1) (4.5 fold), and reduced glutathione (1.3 fold), while the activity of the inhibitor protein increased 2 fold. When the PTU treated rats were challenged with AAP, transaminases were not elevated, and only sporadic areas of necrosis were detected by histological examination of the liver tissue. In contrast to the 12 day treatment with PTU the 3 day treatment had no protection against AAP. No histological evidence of protection was manifested and the transaminases were not different from AAP treated controls. Most of the protective metabolites were depleted. 4 Our findings suggest that PTU‐induced increased concentration of inhibitor protein and GSH, are contributing factors in the prevention of hepatotoxicity by maintaining hepatic m‐ATP levels and reducing the harmful effect of the toxic metabolite of AAP.

[1]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[2]  G. Ellman,et al.  Tissue sulfhydryl groups. , 1959, Archives of biochemistry and biophysics.

[3]  M. E. Pullman,et al.  A NATURALLY OCCURRING INHIBITOR OF MITOCHONDRIAL ADENOSINE TRIPHOSPHATASE. , 1963, The Journal of biological chemistry.

[4]  H. Adam.  Adenosine-5′-triphosphate: Determination with Phosphoglycerate Kinase , 1965 .

[5]  E. Baginski,et al.  Determination of phosphate: Study of labile organic phosphate interference , 1967 .

[6]  L. Horstman,et al.  Partial Resolution of the Enzymes Catalyzing Oxidative Phosphorylation XXII. INTERACTION BETWEEN MITOCHONDRIAL ADENOSINE TRIPHOSPHATASE INHIBITOR AND MITOCHONDRIAL ADENOSINE TRIPHOSPHATASE , 1970 .

[7]  E. Engvall,et al.  Enzyme-linked immunosorbent assay, Elisa. 3. Quantitation of specific antibodies by enzyme-labeled anti-immunoglobulin in antigen-coated tubes. , 1972, Journal of immunology.

[8]  B. Brodie,et al.  ACETAMINOPHEN-INDUCED HEPATIC NECROSIS. III. CYTOCHROME P-450-MEDIATED COVALENT BINDING IN VITRO , 1973 .

[9]  B B Brodie,et al.  Acetaminophen-induced hepatic necrosis. IV. Protective role of glutathione. , 1973, The Journal of pharmacology and experimental therapeutics.

[10]  S. Thorgeirsson,et al.  Acetaminophen-induced hepatic necrosis. VI. Metabolic disposition of toxic and nontoxic doses of acetaminophen. , 1974, Pharmacology.

[11]  R. Barbour,et al.  Purification and properties of ATPase inhibitor from rat liver mitochondria. , 1976, Biochimica et biophysica acta.

[12]  P. Pedersen,et al.  Preparation and characterization of mitochondria and submitochondrial particles of rat liver and liver-derived tissues. , 1978, Methods in cell biology.

[13]  S. Chan,et al.  Use of antibodies for studying the sidedness of membrane components. , 1979, Methods in enzymology.

[14]  D. Roos,et al.  The protective role of glutathione , 1979 .

[15]  A. Pitotti,et al.  F1-ATPase from different submitochondrial particles. , 1979, Biochimica et biophysica acta.

[16]  N. Kaplowitz,et al.  Propylthiouracil. A substrate for the glutathione S-transferases that competes with glutathione. , 1980, The Journal of biological chemistry.

[17]  K. Raheja,et al.  Mechanism of the protective effect of propylthiouracil against acetaminophen (Tylenol) toxicity in the rat. , 1980, Gastroenterology.

[18]  L. Theodorsen,et al.  Experiences with the Scandinavian recommended methods for determinations of enzymes in blood. A report by the Scandinavian Committee on Enzymes (SCE) , 1981, Scandinavian journal of clinical and laboratory investigation.

[19]  N. Kaplowitz,et al.  Direct protection against acetaminophen hepatotoxicity by propylthiouracil. In vivo and in vitro studies in rats and mice. , 1981, The Journal of clinical investigation.

[20]  K. Raheja,et al.  Protective effect of propylthiouracil independent of its hypothyroid effect on acetaminophen toxicity in the rat. , 1982, The Journal of pharmacology and experimental therapeutics.

[21]  K. Raheja,et al.  Prevention of acetaminophen hepatotoxicity by propylthiouracil in the glutathione depleted rat. , 1983, Comparative biochemistry and physiology. C, Comparative pharmacology and toxicology.

[22]  A. Y. Lu,et al.  N-acetyl-p-benzoquinone imine: a cytochrome P-450-mediated oxidation product of acetaminophen. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[23]  B. Mannervik,et al.  2-Propylthiouracil does not replace glutathione for the glutathione transferases. , 1984, The Journal of biological chemistry.

[24]  K. Raheja,et al.  Inhibitory effect of propylthiouracil-induced hypothyroidism in rat on oxidative drug metabolism. , 1985, Comparative biochemistry and physiology. C, Comparative pharmacology and toxicology.

[25]  P. Pedersen,et al.  Regulation of the mitochondrial ATP synthase/ATPase complex. , 1986, Archives of biochemistry and biophysics.

[26]  B. Chernyak,et al.  The effect of the natural protein inhibitor on H+‐ATPase in hepatoma 22a mitochondria , 1987, FEBS letters.

[27]  William Rouslin.  Factors affecting the loss of mitochondrial function during zero-flow ischemia (autolysis) in slow and fast heart-rate hearts. , 1988, Journal of molecular and cellular cardiology.

[28]  William Rouslin,et al.  Factors affecting the reactivation of the mitochondrial adenosine 5'-triphosphatase and the release of ATPase inhibitor protein during and following the reenergization of mitochondria from ischemic cardiac muscle. , 1989, Archives of biochemistry and biophysics.

[29]  M. Tirmenstein,et al.  Subcellular binding and effects on calcium homeostasis produced by acetaminophen and a nonhepatotoxic regioisomer, 3'-hydroxyacetanilide, in mouse liver. , 1989, The Journal of biological chemistry.

[30]  J. Farber,et al.  The killing of cultured hepatocytes by N-acetyl-p-benzoquinone imine (NAPQI) as a model of the cytotoxicity of acetaminophen. , 1991, Biochemical pharmacology.

[31]  A. Mclean,et al.  Effect of paracetamol on mitochondrial membrane function in rat liver slices. , 1991, Biochemical pharmacology.

[32]  J. Noordhoek,et al.  Depletion of ATP but not of GSH affects viability of rat hepatocytes. , 1992, European journal of pharmacology.

[33]  B. Trumpower,et al.  Isolation and characterization of COX12, the nuclear gene for a previously unrecognized subunit of Saccharomyces cerevisiae cytochrome c oxidase. , 1992, The Journal of biological chemistry.

[34]  P. Mohanty,et al.  pH dependent conformational changes modulate functional activity of the mitochondrial ATPase inhibitor protein. , 1993, Biochemical and biophysical research communications.

[35]  A. Mclean,et al.  Adenosine triphosphate (ATP) levels in paracetamol-induced cell injury in the rat in vivo and in vitro. , 1995, Toxicology.

[36]  M. Soledad García,et al.  Kinetic determination of carbimazole, methimazole and propylthiouracil in pharmaceuticals, animal feed and animal livers. , 1995, The Analyst.

[37]  I..,et al.  Isolation and Characterization of COX 12 , the Nuclear Gene for a Previously Unrecognized Subunit of Saccharomyces cerevisiae Cytochrome c Oxidase * , 2022 .