Temporal variation in hepatotoxicity and metabolism of acetaminophen in mice
Introduction
It has been well accepted that biological response to drugs and chemicals may not be constant throughout a day (Halberg, 1969, Moore Ede, 1973, Reinberg, 1979). Conceptually such variations may be dependent on changes in the sensitivity of target organs to a particular agent, on alterations in drug disposition including metabolism and excretion, and/or on rhythmic fluctuations in the availability of endogenous substances such as glutathione (GSH), of which hepatic concentrations have been shown to exhibit a circadian rhythmicity (Jaeger et al., 1973, Schnell et al., 1984).
Acetaminophen (APAP), a widely used analgesic–antipyretic, is safe at therapeutic doses, but upon a large overdose severe hepatotoxicity may result in experimental animals and humans. At low dose levels APAP is detoxified mainly through sulfation and glucuronidation in animals (Jollow et al., 1974). Following an overdose, however, APAP is increasingly metabolized into a reactive metabolite, N-acetyl-p-benzoquinonimine (NAPQI), by cytochrome P450-mediated reactions, which is normally detoxified by conjugation with hepatic GSH. When the generation of reactive metabolites exceeds the availability of hepatic GSH for conjugation, covalent binding of the reactive metabolite to macromolecules may result, an event that correlates with hepatic necrosis. Considering the rhythmicity of hepatic GSH level and its critical role in detoxification of APAP, it is suspected that there would be time-dependent variations in APAP-induced effects. In fact a circadian rhythm in APAP-induced lethality was described in mice (Schnell et al., 1983, Schnell et al., 1984).
The present study was designed to further investigate the temporal variation in response of animals to APAP. We have examined various factors associated with metabolic activation or detoxification of APAP to determine the mechanism responsible for the time-dependent fluctuations in hepatotoxicity and metabolism of this drug.
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Animals and treatments
Adult ICR male mice (Animal Breeding Center, Seoul National University) weighing 30–35 g were used throughout the study. Animals were housed in temperature (22±2°C) and humidity (55±5%) controlled rooms with a 12-h light:dark cycle (light, 08:00–20:00 h). Regular lab chow and tap water were allowed ad libitum until drug administration. Animals were injected with APAP (400 mg/kg, i.p.) either at 08:00, 14:00 or 20:00 h. APAP was injected as a saturated solution in saline (20 ml/kg), prepared by
Variations in APAP-induced hepatotoxicity
Mice were injected with a single dose of APAP (400 mg/kg, i.p.) at 08:00, 14:00 or 20:00 h, and the resulting hepatotoxicity was measured 24 h following each treatment (Table 1). APAP, at the dose used, was not hepatotoxic when administered either at 08:00 or 14:00 h, as measured by changes in serum AST and ALT activities. However, an identical dose of this analgesic given at 20:00 h resulted in a remarkable elevation in these parameters, more than 10- and 50-fold increases in AST and ALT,
Discussion
In the present study an intraperitoneal dose of APAP (400 mg/kg) was administered to mice at 08:00, 14:00 or 20:00 h. Treatment of animals with APAP at 08:00 or 14:00 h did not affect the serum AST/ALT activities, but an identical dose of APAP given at 20:00 h elevated the parameters for hepatotoxicity measured in this study significantly. Similarly, hepatic G-6-Pase activity was depressed by the APAP administration made at 20:00 h only. Among the time points examined, a significantly higher
Acknowledgements
This work was supported by the Korea Science and Engineering Foundation (KOSEF) through the Research Center for New Drug Development (RCNDD) at Seoul National University.
References (30)
- et al.
Ethoxy-, pentoxy- and benzylphenoxazones and homologues: a series of substrates to distinguish between different induced cytochromes P-450
Biochem. Pharmacol.
(1985) Determination of glutathione and glutathione disulfide using glutathione reductase and 2-vinylpyridine
Anal. Biochem.
(1980)- et al.
Circadian variation of hepatic UDP-glucuronic acid and the glucuronidation of xenobiotics in mice
Toxicol. Lett.
(1991) - et al.
Role of CYP2E1 in the hepatotoxicity of acetaminophen
J. Biol. Chem.
(1996) - et al.
Protein measurement with Folin-phenol reagent
J. Biol. Chem.
(1951) - et al.
The toxicity of acetaminophen and N-acetyl-p-benzoquinone imine in isolated hepatocytes is associated with thiol depletion and increased Ca2+
J. Biol. Chem.
(1985) - et al.
The carbon monoxide binding pigment of liver microsomes. I. Evidence for its hemoprotein nature
J. Biol. Chem.
(1964) - et al.
Acetaminophen activation by human liver cytochromes P450IIE1 and P450IA2
Arch. Biochem. Biophys.
(1989) Advances in clinical chronophamacology
Trends Pharmacol. Sci.
(1979)- et al.
Oxidation of acetaminophen to N-acetyl-p-aminobenzoquinone imine by human CYP3A4
Biochem. Pharmacol.
(1993)
Differences in the potentiation of carbon tetrachloride in rats by ethanol and isopropanol pretreatment
Toxicol. Appl. Pharmacol.
Chronobiology
Ann. Rev. Physiol.
Paracetamol metabolism following overdosage: application of high performance liquid chromatography
J. Pharm. Pharmacol.
Diurnal variation of hepatic glutathione concentration and its correlation with 1,1-dichloroethylene inhalation toxicity in rats
Res. Commun. Chem. Pathol. Pharmacol.
Acetaminophen-induced hepatic necrosis
Pharmacology
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