Therapeutic effect of S-allylmercaptocysteine on acetaminophen-induced liver injury in mice
Introduction
Since ancient times, garlic (Allium sativum L.) has been used world-wide as a food and a folk medicine. In recent years, many studies have shown that garlic possesses antimicrobial (Cavallito and Bailey, 1944), antithrombotic (Ariga et al., 1981), antitumor (Sumiyoshi and Wargovich, 1989), antihyperlipidemic Kamanna and Chandrasekhara, 1982, Lau et al., 1987, Steiner et al., 1996, antioxidant Horie et al., 1989, Ide et al., 1996, Imai et al., 1994 and hepatoprotective Hikino et al., 1986, Wang et al., 1996 properties.
S-allylmercaptocysteine is one of the water-soluble organosulfur compounds in the aged garlic extract obtained by ethanol extraction of sliced garlic bulbs. S-allylmercaptocysteine has been shown to inhibit cell proliferation (Sigounas et al., 1997) and to have radical scavenging (Imai et al., 1994) and intraocular pressure-lowering effects (Chu et al., 1999). Moreover, several investigators, including our group, have demonstrated that S-allylmercaptocysteine pretreatment protects the liver against a number of hepatotoxicants, such as acetaminophen, carbon tetrachloride and d-galactosamine Hikino et al., 1986, Nakagawa et al., 1989, Sumioka et al., 1998.
Acetaminophen is widely used as an analgesic and antipyretic drug. Although acetaminophen is harmless at therapeutic doses, it can produce hepatic injury in both human and experimental animals when given in high doses. After therapeutic doses, it is biotransformed and eliminated as non-toxic glucuronic acid and sulfate conjugates Black, 1980, Pacifici et al., 1988. Only a small proportion of acetaminophen is converted to N-acetyl-p-benzoquinoneimine (NAPQI) (Dahlin et al., 1984), a cytochrome P450 (CYP)-mediated intermediate metabolite, which is normally detoxified by conjugation with GSH. However, after high doses of acetaminophen, the capacity for its removal by hepatic conjugation with glucuronide and sulfate is exceeded, and more of the reactive metabolite NAPQI is formed. Consequently, more NAPQI is conjugated with GSH, and when the hepatic GSH is depleted, more NAPQI will bind covalently to cellular macromolecules Jollow et al., 1973, Potter and Hinson, 1986. This adduction of proteins by NAPQI is thought to lead to liver injury, and to play a key role in triggering heat shock protein (HSP) induction (Salminen et al., 1998). Mice and hamsters have been shown to be very sensitive to the hepatotoxic effects of acetaminophen, developing fulminant centrilobular necrosis similar to that observed in the human (Hinson, 1980).
Protection against acetaminophen-induced liver injury can be achieved by various mechanisms. Cysteine prodrugs, including N-acetylcysteine, the most widely used antidote for acetaminophen overdose, have been reported to protect the liver against acetaminophen-induced injury. The mechanism responsible for this protection may be metabolism of these prodrugs to l-cysteine, which is incorporated into hepatic GSH Corcoran and Wong, 1986, Hazelton et al., 1986a, Lauterburg et al., 1983, Miners et al., 1984, Roberts et al., 1987. Moreover, several CYP enzymes have been reported to play important roles in the bioactivation of acetaminophen to NAPQI Harvison et al., 1988, Morgan et al., 1983, Patten et al., 1993, Raucy et al., 1989. Studies on CYP2E1 knockout mice have made it clear that CYP2E1 is the most important factor in acetaminophen bioactivation Lee et al., 1996, Zaher et al., 1998. In addition, CYP2E1 inhibitors such as diallyl sulfide, phenethyl isothiocyanate and 2-(allylthio)pyrazine protect the liver against acetaminophen-induced injury Kim et al., 1997, Li et al., 1997, Wang et al., 1996. This evidence supports the crucial role of the CYP2E1 enzyme in acetaminophen-induced liver damage. Furthermore, results of recent studies suggest that acetaminophen-induced liver injury is also caused by cellular oxidative stress, resulting in hepatic lipid peroxidation Albano et al., 1983, Amimoto et al., 1995, Wendel et al., 1979. Several antioxidants and antioxidative enzymes, such as coenzyme Q10, α-tocopherol, ascorbic acid and superoxide dismutase, have been shown to prevent acetaminophen-induced liver injury in vivo Amimoto et al., 1995, Mitra et al., 1991, Nakae et al., 1990. Finally, enhancing the capacity for conjugation with glucuronic acid may protect against acetaminophen-induced liver injury, as shown in an experiment that used butylated hydroxyanisole (Hazelton et al., 1986b).
In a previous study, we demonstrated that S-allylmercaptocysteine pretreatment before acetaminophen administration protected mice against acetaminophen-induced liver injury Nakagawa et al., 1989, Sumioka et al., 1998. Considering the clinical possibility for the use of S-allylmercaptocysteine for acetaminophen overdose, it is very important to examine whether S-allylmercaptocysteine treatment after acetaminophen overdose can relieve hepatotoxicity. In this study, we, therefore, examined the therapeutic effect of S-allylmercaptocysteine against acetaminophen-induced hepatotoxicity. Since the mechanisms responsible for this hepatoprotection may be inhibition of CYP2E1 activity (Sumioka et al., 1998), we assumed that formation of the CYP-mediated reactive metabolite, NAPQI and subsequent acetaminophen arylation of protein would be decreased by S-allylmercaptocysteine treatment. However, we had no data to support our assumption. To clarify this, we examined whether S-allylmercaptocysteine treatment affected acetaminophen-induced hepatic-inducible 70-kDa heat shock protein (HSP70i) induction. This induction is strongly associated with the hepatic intralobular sites of acetaminophen arylation of protein (Salminen et al., 1997a).
Section snippets
Chemicals
S-allylmercaptocysteine was synthesized as described previously (Hikino et al., 1986). Acetaminophen, 2-vinylpyridine and phenylmethylsulfonyl fluoride were purchased from Wako (Osaka, Japan). Glutathione reductase, isocitric dehydrogenase and N-acetylcysteine were purchased from Sigma (St. Louis, MO, USA). NADPH and NADP were purchased from Boehringer Mannheim Biochemicals (Mannheim, Germany). N-nitrosodimethylamine was purchased from Tokyo Kasei Industry (Tokyo, Japan).
Effect of S-allylmercaptocysteine on acetaminophen-induced hepatotoxicity
The effect of S-allylmercaptocysteine on acetaminophen-induced mortality is shown in Table 1. Treatment with only the vehicle 0.5 h after acetaminophen administration gave a mortality of 43% by 48 h after acetaminophen administration. S-allylmercaptocysteine treatment 0.5 h after acetaminophen administration completely blocked acetaminophen-induced mortality. However, S-allylmercaptocysteine treatment 1 h after acetaminophen administration did not improve acetaminophen-induced mortality.
Discussion
In a previous study, we demonstrated that S-allylmercaptocysteine treatment (once a day for 2 days) before acetaminophen administration dramatically protects mice against acetaminophen-induced liver injury Nakagawa et al., 1989, Sumioka et al., 1998. In the current study, we examined the therapeutic effect of S-allylmercaptocysteine given after acetaminophen and its potential application to clinical therapy.
A single dose of S-allylmercaptocysteine 0.5 h after acetaminophen administration
Acknowledgements
We are grateful to Professor K. Sato, Department of Molecular Biology, School of Life Science, Tottori University, for his helpful discussion.
References (51)
- et al.
Paracetamol-stimulated lipid peroxidation in isolated rat and mouse hepatocytes
Chem.-Biol. Interact.
(1983) - et al.
Acetaminophen-induced hepatic injury in mice: the role of lipid peroxidation and effects of pretreatment with coenzyme Q10 and α-tocopherol
Free Radical Biol. Med.
(1995) - et al.
Platelet aggregation inhibitor in garlic
Lancet
(1981) Acetaminophen hepatotoxicity
Gastroenterology
(1980)Determination of glutathione and glutathione disulfide using glutathione reductase and 2-vinylpyridine
Anal. Biochem.
(1980)- et al.
Effects of butylated hydroxyanisole on acetaminophen hepatotoxicity and glucuronidation in vivo
Toxicol. Appl. Pharmacol.
(1986) - et al.
Inhibition of cytochrome P450 2E1 expression by 2-(allylthio)pyrazine, a potential chemoprotective agent: hepatoprotective effects
Biochem. Pharmacol.
(1997) - et al.
Effect of an odor-modified garlic preparation on blood lipids
Nutr. Res.
(1987) - et al.
Role of CYP2E1 in the hepatotoxicity of acetaminophen
J. Biol. Chem.
(1996) - et al.
Effects of phenethyl isothiocyanate on acetaminophen metabolism and hepatotoxicity in mice
Toxicol. Appl. Pharmacol.
(1997)
Protein measurement with the folin phenol reagent
J. Biol. Chem.
Mechanism of action of paracetamol protective agents in mice in vivo
Biochem. Pharmacol.
Comparison of six rabbit liver cytochrome P-450 isozymes in formation of a reactive metabolite of acetaminophen
Biochem. Biophys. Res. Commun.
Sulphation and glucuronidation of paracetamol in human liver: assay conditions
Biochem. Pharmacol.
Acetaminophen activation by human liver cytochromes P450 IIE1 and P450 IA2
Arch. Biochem. Biophys.
A double-blind crossover study in moderately hypercholesterolemic men that compared the effect of aged garlic extract and placebo administration on blood lipids
Am. J. Clin. Nutr.
Protective effects of garlic and related organosulfur compounds on acetaminophen-induced hepatotoxicity in mice
Toxicol. Appl. Pharmacol.
Acute paracetamol intoxication of starved mice leads to lipid peroxidation in vivo
Biochem. Pharmacol.
Protection against acetaminophen toxicity in CYP1A2 and CYP2E1 double-null mice
Toxicol. Appl. Pharmacol.
Allicin, the antibacterial principle of Allium sativum. I. Isolation, physical properties and antibacterial action
J. Am. Chem. Soc.
Intraocular pressure lowering by S-allylmercaptocysteine in rabbits
J. Ocul. Pharmacol. Ther.
Role of glutathione in prevention of acetaminophen-induced hepatotoxicity by N-acetyl-l-cysteine in vivo: studies with N-acetyl-d-cysteine in mice
J. Pharmacol. Exp. Ther.
N-acetyl-p-benzoquinone imine: a cytochrome P-450-mediated oxidation product of acetaminophen
Proc. Natl. Acad. Sci. U. S. A.
Cytochrome P-450 isozyme selectivity in the oxidation of acetaminophen
Chem. Res. Toxicol.
Effects of cysteine pro-drugs on acetaminophen-induced hepatotoxicity
J. Pharmacol. Exp. Ther.
Cited by (45)
Phytochemical composition, isolation and hepatoprotective activity of active fraction from Veronica ciliata against acetaminophen-induced acute liver injury via p62-Keap1-Nrf2 signaling pathway
2019, Journal of EthnopharmacologyCitation Excerpt :Some studies began to found new agents from other sources, such as the traditional medicinal plants with some active properties, which could relieve the liver injury induced by APAP and possess huge promise for being developed into drugs for treatment of the liver diseases caused by APAP. ( Luan et al., 2016; Shao et al., 2017; Sumioka et al., 2001). Veronica ciliata Fisch.
Effect of chrysin on the formation of N-acetyl-p-benzoquinoneimine, a toxic metabolite of paracetamol in rats and isolated rat hepatocytes
2019, Chemico-Biological InteractionsCitation Excerpt :Number of in vitro studies reported that chrysin has a potent inhibitory effect on CYP2E1, CYP3A4 and CYP1A2 [53–59]. Previous studies revealed that aminotriazole [60], ozagrel hydrochloride [61], ornithogalum saundersiae [62], dioscin [63], lupeol [64], phyllanthus urinaria extract [65], α- and β-amyrin [66], lipopolysaccharide-binding protein [67], moutan cortex extract [68], baicalin [69], S-allylmercaptocysteine [70], berberine [71], propylene glycol [72], ebselen [73], allylsulfide, allylmercaptan, allylmethylsuhide [74], diallyl sulfone [75], fucoidan [76] and genistein [77] exhibited hepatoprotective activity against paracetamol-induced hepatotoxicity by reducing the formation of NAPQI via CYP2E1 inhibition. In the present study, chrysin increased the Cmax of paracetamol by 1.74-folds.
Anti-cancer activities of S-allylmercaptocysteine from aged garlic
2019, Chinese Journal of Natural MedicinesPotential protective role of nitric oxide and Hsp70 linked to functional foods in the atherosclerosis
2017, Clinica e Investigacion en ArteriosclerosisAged garlic extract enhances heme oxygenase-and glutamate-cysteine ligase modifier subunit expression via the nuclear factor erythroid 2-related factor 2-antioxidant response element signaling pathway in human endothelial cells
2016, Nutrition ResearchCitation Excerpt :AGE is also less irritating and has no toxic side effects [7,8]. AGE and its components have been reported to possess beneficial pharmacological activities, such as anti-stress [9], anti-fatigue [10], immunomodulation [11], and cardiovasculo- [12–14] and hepato-protective [15] effects. Cardiovascular disease is characterized by various events, including an increase in oxidative stress [16].
- 1
Tel.: +81-826-45-2331; fax: +81-826-45-4351.