Chlorpropham induces mitochondrial dysfunction in rat hepatocytes
Introduction
Chlorpropham (isopropyl N-(3-chlorophenyl)carbamate; CIPC) is used to control weeds in various food and non-food crops, and as a sprouting suppressant and/or for a post-harvest treatment of potatoes during storage and/or transport. Even if consumption in the diet is low, the potential toxicity of CIPC has been studied in vivo and in vitro to assess its various toxicologic properties. Although CIPC was not potently toxic in genotoxic, mutagenic, and immunotoxic studies and in some short- and/or long-term dietary studies (Van Esch and Kroes, 1972, Larson et al., 1960, Boyd and Carsky, 1969, Van Engelen, 2001), the liver, kidney, spleen and erythrocytes were identified as targets of CIPC-induced toxicity in rats and mice. Oral administration of CIPC to rats or mice caused dose-related increases in liver and spleen weight, accompanied by the acceleration of hematopoiesis and/or hemosiderosis in both organs. The serum concentrations of bilirubin and cholesterol, as parameters of hepatic damage, in rats were significantly increased at a high dose (2200 mg/kg per day) in a 28-day study (Van Engelen, 2001). The hepatic damage in mice was accompanied by focal necrosis (Fujitani et al., 2000). In the kidney, intracytoplasmic hyaline resportion bodies were found in the tubules, with the effect being more pronounced in males than in females (Fujitani et al., 1997, Fujitani et al., 2000, Van Engelen, 2001). In addition, it has been reported that CIPC is well absorbed from the gastrointestinal tract when administered via the oral route in rats and mice, undergoes extensive metabolism, and is excreted in the urine and that the metabolism of CIPC proceeds either via aromatic 4-hydroxylation followed by conjugation of the intermediate with glucuronic acid and sulfate as a major pathway, or via oxidation of the isopropyl side-chain and carbamate hydrolysis, followed by conjugation of these intermediates as minor pathways (Fang et al., 1974, Carrera et al., 1998).
The metabolic pathway and toxic effects of CIPC have been studied in vivo and/or in vitro (Fang et al., 1974, Carrera et al., 1998), whereas the relationship between metabolism and toxicity has not been clarified. The freshly isolated rat hepatocyte system which retains intact membranes and has high levels of various drug-metabolizing enzymes and their cofactors associated with Phase I and Phase II reactions is useful for the study of uptake, intracellular target sites, and temporal sequences leading to cell injury by chemicals and/or their metabolites via biotransformation (Moldéus et al., 1978). In the present study, we investigated the metabolism and cytotoxic effects of CIPC on isolated rat hepatocytes.
Section snippets
Materials
The chemical compounds used were obtained from the following companies: CIPC (purity of >99.7%) from Hodogaya Chemical Industry (Tokyo, Japan); 3-chloroaniline (3CA) and 3-chloro-4-hydroxyaniline from Tokyo Kasei Kogyo Co., Ltd. (Tokyo); β-glucuronidase, sulfatase (type VI, β-glucuronidase-free Aerobacter aerogenes), N-(2-hydroxyethyl)-piperazine-N-(2-ethanesulfonic acid) (HEPES), 2-diethylaminoethyl-2,2-diphenylvalerate hydrochloride (SKF-525A) and bovine serum albumin from Sigma Chemical Co.
Toxic effects of CIPC on rat hepatocytes
The exposure of rat hepatocytes to CIPC caused a concentration (0.25–1.0 mM) and time-dependent (0–3 h) cell death accompanied by a depletion of intracellular ATP and total adenine nucleotides (Fig. 2). CIPC at a concentration of 1.0 mM elicited a rapid depletion of total nucleotide pools with abrupt lethality, whereas the loss of ATP was reflected by concomitant increases in the levels of ADP and AMP (data not shown). The concentrations of malondialdehyde, an index of lipid peroxidation, in
Discussion
The results obtained in the present study show that in isolated rat hepatocytes, CIPC induced a concentration- and time-dependent loss of cell viability, followed by decreases in intracellular levels of ATP and total adenine nucleotide pools. The cytotoxicity of CIPC may depend on the initial concentration rather than on that of its major intermediate 4OH-CIPC, because (a) the pretreatment of hepatocytes with the inhibitor of microsomal monooxygenase enhanced CIPC-induced cytotoxicity
References (29)
- et al.
Metabolic studies of -labelled propham and chlorpropham in the female rats
Pest. Biochem. Physiol.
(1974) - et al.
Hemotoxicity of chlorpropham (CIPC) in F344 rats
Toxicology
(1997) - et al.
Subchronic toxicity of chlorpropham (CIPC) in ICR mice
Food Chem. Toxicol.
(2000) - et al.
Interaction of butylated hydroxyanisole with mitochondrial oxidative phosphorylation
Biochem. Pharmacol.
(1992) Determination of pyridine dinucleotides in cell extracts by high-performance liquid chromatography
J. Chromatog.
(1981)- et al.
Chronic toxicologic studies on isopropyl-N-(3-chlorophenyl)carbamate (CIPC)
Toxicol. Appl. Pharmacol.
(1960) - et al.
Protein measurement with the Folin phenol reagent
J. Biol. Chem.
(1951) - et al.
Isolation and use of liver cells
Methods Enzymol.
(1978) - et al.
Mechansim of p-hydroxybenzoate ester-induced mitochondrial dysfunction and cytotoxicity in rat hepatocytes
Biochem. Pharamcol.
(1998) - et al.
J. Biol. Chem.
(1990)
Uncoupling properties of a chlorophenol series on Acer cell suspensions: a QSAR study
Ecotoxicol. Environ. Saf.
Role of redox cycling and lipid peroxidation in bipyridyl herbicide cytotoxicity. Studies with a compromised isolated hepatocyte model system
Biochem. Pharmacol.
Potentiation of oxidative cell injury in hepatocytes which have accumulated Ca2+
J. Biol. Chem.
Long-term toxicity studies of chlorpropham and propham in mice and hamsters
Food Cosmet. Toxicol.
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