Induction of hepatic microsomal cytochrome P-450 by mirex and kepone.

The chlorinated insecticides, mirex and Kepone, pose a threat to human health as a consequence of their pollution of the environment. We investigated their potential to affect synergistically the toxicity of other xenobiotics and the pharmacological function of drugs by induction of hepatic microsomal enzymes. Male rats were induced by ip injection of mirex (50 or 5 mg/kg/day for 5 days) or Kepone (10 or 1 mg/kg/day for 5 days). Metabolic activity was tested with warfarin and biphenyl using high-performance liquid chromatographic assays. The high doses of both compounds induced cytochrome P-450 with absorbance bands (reduced, CO complex) at 449 nm. Cytochrome concentrations were enhanced twofold relative to controls. Mirex resembled 3-methylcholanthrene and benzo[a]pyrene by inducing formation of 6-hydroxywarfarin but differed in not inducing 8-hydroxywarfarin. Kepone resembled phenobarbital in inducing 7-hydroxywarfarin but differed in its effects on the other metabolites. The low dose of mirex induced higher amounts of 4′-hydroxywarfarin than did the high dose. The metabolite profiles with high and low doses of Kepone also showed marked variations from one another. Mirex and Kepone are carcinogenic in rats and mice but, in contrast to the polycyclic aromatic carcinogens, do not markedly enhance the activity of microsomal biphenyl 2-hydroxylase relative to biphenyl 4-hydroxylase. We conclude that mirex and Kepone induce hepatic mixed-function oxidase profiles which differ from one another and from the classical inducers, phenobarbital and 3-methylcholanthrene. Mirex apparently only induces one of the enzymes induced by 3-methylcholanthrene. The enzyme profiles arising from the insecticides are dose dependent and will thus potentiate qualitatively differing effects depending on the level of ingestion.     

In vitro inhibition of mouse hepatic mixed-function oxidase enzymes by helenalin and alantolactone

The sesquiterpene lactones (STL) helenalin and alantolactone were effective in vitro inhibitors of the mouse hepatic microsomal mixed-function oxidase (MFO) enzymes, aminopyrine demethylase (APD), aniline hydroxylase (ANH) and 7-ethoxyresorufin deethylase (ERD). Helenalin and alantolactone concentrations of 0.5 mM produced a 50-60% inhibition of APD and ERD, and a 20-30% inhibition of ANH. An increase in substrate (aminopyrine) concentration from 0.5 to 25 mM decreased STL inhibition of APD by 12-32%. APD was also inhibited at low aminopyrine concentrations (0.5 mM) by the helenalin derivative 2,3,11,13-tetrahydrohelenalin (tetrahydrohelenalin). The STL produced type I binding spectra with oxidized microsomes; Ks values for helenalin and alantolactone were 161 and 9 microM respectively. These results suggest that STL inhibition of the MFO system results, in part, from STL binding to the substrate-binding site of cytochrome P-450. It has been reported that the irreversible alkylation of protein cysteinyl residues is responsible for STL inhibition of several different enzymes, and second-order rate constants for the reaction of helenalin and alantolactone with glutathione were 25.1 and 1.80 mM-1.hr-1 respectively. Tetrahydrohelenalin did not react with glutathione. However, the subsequent addition of 3.0 mM thiols, i.e. L-cysteine, N-acetylcysteine or glutathione, to STL-treated (0.5 mM) microsomes reversed helenalin and alantolactone inhibition of APD and ERD by 50-80%. The ability of thiols to reverse STL inhibition of APD was decreased 20-43% by the coincubation of STL and microsomes with an NADPH-generating system. In addition, established effects of sulfhydryl-reactive compounds on the MFO system, i.e. inhibition of NADPH-cytochrome c reductase and conversion of cytochrome P-450 to cytochrome P-420, were not observed after addition of helenalin (1.0 mM) or alantolactone (0.5 mM) to mouse hepatic microsomes. These results suggest that STL inhibition of MFO enzymes may not be dependent upon the reactivity of the STL towards sulfhydryl groups. Instead, we suggest that STL binding to the substrate-binding site of cytochrome P-450 and subsequent metabolism of the STL may contribute to inhibition of the MFO system.     

Species differences in stimulation of intestinal and hepatic microsomal mixed-function oxidase enzymes

The effect of intraperitoneal (i.p.) administration of phenobarbital (PB) or 3-methylchol-anthrene (3-MC) on some mixed-function oxidase (MFO) enzymes was studied in small intestine and liver of male rats, mice, guinea pigs and rabbits. PB treatment enhanced intestinal and 7-ethoxycoumarin deethylase activities in the mouse and rat, whereas benzo[a]pyrene hydroxylase (AHH) activity was increased only in the mouse. Ethylmorphine demethylase and aniline hydroxylase activities in small intestine were not stimulated by PB in any species. Administration of 3-MC increased the activity of intestinal AHH in rat, mouse and guinea pig, but intestinal 7-ethoxycoumarin deethylase activity was elevated only in the rat. The guinea pig and mouse intestinal ethoxycoumarin deethylase activity was inhibited by 3-MC treatment. None of the enzymes tested in rabbit intestine was induced by PB or 3-MC. The hepatic activities of ethylmorphine demethylase, aniline hydroxylase, 7-ethoxycoumarin deethylase and AHH, and the cytochrome P-450 content were increased by PB in all species. In contrast, 3-MC enhanced hepatic aniline hydroxylase and AHH activities in rats, mice and guinea pigs, and hepatic 7-ethoxycoumarin deethylase activity in mice and rats. In rabbits, these hepatic enzymes were inhibited by 3-MC pretreatment. The hepatic cytochrome P-450 absorption spectra was shifted to 448 nm in all species. These results suggest that there are differences in induction of intestinal and hepatic MFO enzymes which are influenced by the type of inducing agent, substrate and animal species used.     

Glutathione and hepatic mixed-function oxidase activity

It is apparent that hepatic GSH may function in drug metabolism not only as a substrate for conjugation but also in regulation of cytochrome P-450 activity. The remarkable aspect of the latter activity is its specificity. Loss of hepatic GSH depresses N-demethylation of DAB while ring hydroxylation is unaffected. On the other hand, the effect is to some degree nonspecific in that control as well as PB- or MC-induced N-demethylation is inhibited. Thus the response may not simply be specific to one isozyme of cytochrome P-450 but may be associated with one aspect of the enzymic activity of several cytochrome P-450 isozymes (i.e., N-demethylation). We have postulated that sensitivity of this activity to lipid peroxidation underlies the relationship to GSH since the tripeptide serves as a major protection against hepatic lipid peroxidation and its consequences. It is as yet not clear as to how or why this particular aspect of P-450 activity is more sensitive to lipid peroxidation than are other activities such as ring hydroxylation. Ongoing investigations include attempts to identify the cytochrome P-450 isozyme(s) which inhibit this response to GSH depletion. GSH-lipid peroxidation relationships have already been reported with isolated hepatocytes, and there may be a possible connection between this and the relative instability of cytochrome P-450 in cultured hepatocytes. Another factor which may be involved is heme oxygenase activity, which is markedly induced in the liver after GSH depletion, after cobalt administration (which also depresses cytochrome P-450 activity), and during incubation of isolated hepatocytes. This enzyme catalyzes the rate-limiting step in heme breakdown and may contribute to the loss of cytochrome P-450 activity associated with GSH depletion.     

Effect of preexposure to kepone on hepatic mixed-function oxidases in the female rat.

The effects of preexposure of female rats to Kepone on hepatic drugmetabolizing enzymes and several other parameters of the multifunction oxidase system were investigated. Animals were exposed to 0, 50, 100, and 150 ppm Kepone in the daily ration for 16 days. There was a dose-related decline in body weight gained to 72, 55, and 22% of the controls for 50, 100, and 150 ppm Kepone, respectively. Liver weight was unaltered at all three levels of dietary Kepone. While hydroxylation of aniline, pentobarbital, and hexobarbital were all enhanced, the increase in metabolism of the latter two substrates was consistently higher than that of controls. Enhanced metabolism of pentobarbital was also evident from the dose-related decrease in the pentobarbital-induced sleeping time. Aminopyrine and ethylmorphine demethylase activities were increased three- to fourfold over the controls. Cytochrome P-450, NADPH-cytochrome c reductase, and aniline binding were all increased, suggesting that there were general increases in the intermediate steps in the electron-transfer system within the multifunction oxidase complex. Cytochrome b₅ was unaltered by Kepone and only a 100-ppm dose elicited a discernible increase in NADPH dehydrogenase activity. These effects in the female rats, along with previously published effects in the male rat, are suggestive of the potential of Kepone to modify hepatic drug-metabolizing function in animals of both sexes.     

Effect of ampicillin on hepatic microsomal mixed-function oxidase system in male mice

The effect of ampicillin [(D)-alpha-aminobenzyl penicillin] administration on the hepatic mixed-function oxidase (MFO) system was studied in male mice. Ampicillin (100 mg/kg, i.p., 3 days) decreased the levels of cytochrome P-450, aminopyrine N-demethylase, acetanilide hydroxylase and cytochrome c-reductase activity significantly. In carbon tetrachloride (CCl4)-pretreated mice, ampicillin increased acetanilide hydroxylation compared with CCl4 treatment alone; however, all other parameters of the MFO system remained unchanged. Ampicillin exhibited type II binding with microsomes (trough at 388 nm, peak at 430 nm). Thus ampicillin acts as an inhibitor of the MFO system.     

Effect of thioacetamide-induced hepatic necrosis on the regioselective metabolism of S-warfarin by rat liver mixed-function oxidase enzymes

The biotransformation of S-warfarin was examined using liver microsomes prepared from rats 6-96 hr after treatment with a necrotizing dose (5.6 mmoles/kg) of thioacetamide. Four catalytically distinct classes of enzyme activity were observed which declined in activity with different half-lives after thioacetamide intoxication. S-Warfarin 7-hydroxylase activity was destroyed with a half-life of 16.6 +/- 3.1 hr. 6-Hydroxylase activity was destroyed with a half-life of 25.3 +/- 3.0 hr. 4'-Hydroxylase activity was destroyed with a half-life of 34.6 +/- 4.8 hr, which paralleled the loss of total hepatic cytochrome P-450 with a half-life of 33.4 +/- 3.6 hr. Production of an unidentified metabolite was not affected by thioacetamide intoxication during the first 48 hr. The ratio of rates of product formation were used as an alternative method to test the homogeneity of distinct enzyme catalytic activities. The ratio of measured responses (e.g. chromatographic peak heights) was used directly to determine the product ratios, provided that the rate of formation of each product was directly proportional to the experimentally measured response for each product. The use of product response ratios to discriminate between catalytic activities was inherently more precise because calibration errors were eliminated. Differences in the rates of destruction of warfarin hydroxylases provided further evidence of the multiplicity of hepatic mixed-function oxidases and suggested topographical differences in their location within the liver lobule.     

Effects of arsenite on hepatic mixed-function oxidase activity in rats.

1. Injection of arsenite (As3+) to control rats results in losses of total hepatic cytochrome P-450 and significant decreases of ethoxycoumarin O-deethylase (ECOD) and ethoxyresorufin O-deethylase (EROD) activities. However, As3+ appears to decrease the activity of these enzymes differentially, with EROD showing greater sensitivity than ECOD. 2. Injection of As3+ to rats treated with phenobarbital and isosafrole significantly decreases the total content of hepatic cytochrome P-450 and various mixed function oxidase (MFO) activities, with the exception of ECOD which appears to be insensitive to As3+. 3. 3-Methylcholanthrene administration apparently protects against the effects of As3+ on the cytochrome P-450 system, since total content of the cytochrome P-450 and various MFO activities were all insensitive to this treatment.     

Effects of the mycotoxins citrinin and ochratoxin a on hepatic mixed-function oxidase and adenosinetriphosphatase in neonatal rats

The effects of citrinin, ochratoxin A, or a combination of the two mycotoxins on the hepatic monooxygenase system and on hepatic and renal adenosinetriphosphatase (ATPase) activities were examined in neonatal rats exposed to a single treatment of one or both toxins. Animals received (po) 25 mg/kg citrinin, 1 mg/kg ochratoxin A, or 25 mg/kg citrinin plus 1 mg/kg ochratoxin A within 24 h of birth. Pups were killed 12 d later. Citrinin or ochratoxin A alone did not affect hepatic ATPase. Renal oligomycin-sensitive Mg2+-ATPase was inhibited to the same degree by ochratoxin A and the combination treatment. A synergistic effect of the two mycotoxins was observed on renal Na+-K+-ATPase. Significant effects, due to the mycotoxin interaction, were also observed on cytochrome P-450 content, NADPH-dependent dehydrogenase, and NADPH-cytochrome c reductase.     

Effects of vitamin A deficiency on hepatic and extrahepatic mixed-function oxidase and epoxide-metabolizing enzymes in guinea pig and rabbit.

Male guinea pigs and male rabbits were fed a vitamin A deficient diet for 9 weeks and for 12 weeks respectively. Hepatic levels of vitamin A were significantly reduced in the vitamin A deficient animals. The activities of some xenobiotic-metabolizing enzymes were measured in the liver, lung and small intestine. Aryl hydrocarbon hydroxylase, aniline hydroxylase, and 7-ethoxycoumarin deethylase activities were decreased in the vitamin A deficient guinea pig liver. However, in the guinea pig small intestine, aniline hydroxylase, 7-ethoxycoumarin deethylase, aminopyrine demethylase, and aryl hydrocarbon hydroxylase specific activities were increased. In rabbits, vitamin A deficiency decreased hepatic aniline hydroxylase and 7-ethoxycoumarin deethylase activities but increased intestinal aminopyrine demethylase activity. Enzyme activities in lung were not altered by vitamin A deficiency in guinea pig or rabbit. Microsomal epoxide hydrase and microsomal supernatant glutathione S-transferase activities in the three tissues of both species were not altered by vitamin A deficiency.     

Effect of sodium fluoride on hepatic microsomal mixed-function oxidase system in newborn and adult male rats

Administration i.p. of sodium fluoride at 2.5 or 5.0 mg/kg induced the mixed-function oxidase system in both newborn and adult rats; at 10.0 mg/kg there was induction in newborn rats and inhibition in adult rats, and at 20.0 mg/kg, there was inhibition in both newborn and adults.     

Induction of the rat hepatic microsomal mixed-function oxidases by cimetidine

The ability of cimetidine to induce the hepatic microsomal mixed-function oxidases was investigated in rats treated orally with the drug at 3 dose levels: 10, 100 and 500 mg/kg. At the highest dose only, cimetidine stimulated the dealkylations of ethoxyresorufin, ethoxycoumarin and pentoxyresorufin but inhibited that of erythromycin and had no effect on the demethylation of dimethylnitrosamine. At the highest dose cimetidine had a small effect on the activation of Glu-P-1 to mutagens in the Ames test but induced proteins recognised in Western blots by antibodies to P450 I A1 and P450 II B1. It is concluded that cimetidine is a weak selective inducer of cytochrome P-450 forms, but at therapeutic doses its inductive effect is most unlikely to be of any clinical or toxicological consequence.     

Inhibition of hepatic mixed-function oxidase activity in vitro and in vivo by various thiono-sulfur-containing compounds.

Ten thiono-sulfur-containing compounds of varying structure were administered by intraperitoneal injection to untreated, phenobarbital-pretreated and 3-methylcholanthrene-pretreated adult male rats. Six hr later, the concentration of hepatic cytochrome P-450 and the ability of the hepatic microsomes to metabolize benzphetamine were examined. In the untreated, phenobarbital-pretreated and 3-methylcholanthrene-pretreated groups, two, four and four compounds, respectively, significantly decreased the concentration of cytochrome P-450 in the hepatic microsomes. A similar effect on benzphetamine metabolism was also seen. When examined 48 hr after the administration of the ten thiono-sulfurcontaining compounds, four, five and seven of the compounds decreased both the levels of hepatic cytochrome P-450 and the rate of benzphetamine metabolism in the untreated, phenobarbital-pretreated and 3-methylcholanthrene-pretreated animals respectively. Eight of the thiono-sulfur-containing compounds were incubated in the presence of NADPH with hepatic microsomes isolated from untreated, phenobarbital-pretreated or 3-methylcholanthrene-pretreated animals. All of the compounds examined significantly decreased the concentration of cytochrome P-450 in the microsomes from each treatment group. Similar reductions in benzphetamine metabolism were also seen. When these same compounds were incubated with microsomes in the absence of NADPH, no significant reduction of cytochrome P-450 or benzphetamine metabolism was seen. When the oxygen analogs of six of the thiono-sulfur compounds were administered in vivo or incubated with hepatic microsomes either in the presence or absence of NADPH, no significant reduction of cytochrome P-450 or benzphetamine metabolism was seen.     

Impairment of hepatic mixed-function oxidase activity by alpha- and beta-naphthylisothiocyanate: relationship to hepatotoxicity.

Since administration of α-naphthylisothiocyanate (ANIT) to rats inhibits hepatic mixed-function oxidase (MFO) activity in vivo and in vitro, the nature of this inhibition was investigated; its role in the pathogenesis of ANIT-cholestasis was also examined. Male rats were treated (150 mg/kg, ip) with ANIT or β-naphthylisothiocyanate (BNIT), a noncholestatic isomer. ANIT was also given to hamsters (a resistant species) and to rabbits (a nonsusceptible species). The animals were killed at different intervals (2–72 hr); MFO activity was compared to plasma biochemical alterations (glutamic-pyruvic transaminase, bilirubin, 5′-nucleotidase) indicative of liver injury. In the rat, reductions in cytochrome P-450, aniline oxidase, and aminopyrine demethylase were evident before the development of hyperbilirubinemia. After 24 hr, cytochrome b₅, total heme, and NADPH-cytochrome c reductase were also reduced, but glucuronyl transferase activity was increased. BNIT caused no elevation in plasma bilirubin or 5′-nucleotidase activity but decreased MFO activity in rats. Reductions in cytochrome P-450 and microsomal metabolism were also observed in hamsters and rabbits treated with ANIT, although only a moderate hyperbilirubinemia was observed in hamsters and none in rabbits. Both ANIT and BNIT caused reductions in cytochrome P-450 when incubated in vitro with microsomes. Loss of cytochrome P-450 due to increased degradation is suggested. We conclude that the metabolic lesion induced by ANIT is primary and does not result from the accumulation of bile acids within the hepatocyte. It appears, however, that this lesion has no causal relationship to the subsequent development of ANIT-induced cholestasis.     

Altered activity of hepatic mixed-function mono-oxygenase enzymes in streptozotocin-induced diabetic rats.

1. In streptozotocin-induced diabetic male rats, hepatic microsomal aminopyrine N-demethylase activity was depressed, whereas aniline hydroxylase activity and cytochrome P-450 content were increased over control values. 2. In diabetic female rats, hepatic microsomal aminopyrine N-demethylase activity, aniline hydroxylase activity, biphenyl 4-hydroxylase activity, and cytochrome P-450 content were increased over control values. 3. Insulin treatment of diabetic male and female rats antagonized all physical and biochemical abnormalities of the diabetic state; 4. Methyl analogues of streptozotocin did not produce a diabetic state when injected into female rats, and resulted in no changes in aminopyrine N-demethylase activity, aniline hydroxylase activity, or cytochrome P-450 content. 5. Insulin treatment of non-diabetic female rats resulted in slight decreases in aminopyrine N-demethylase and aniline hydroxylase activities, but no changes in cytochrome P-450 content. These observations suggest that insulin primarily influences drug metabolism of diabetic animals through correction of the insulin-deficient diabetic state.     

The effects of mirex on the neonatal rat lens in vitro, with a comparison to kepone

Lenses were removed from 12-day-old rats and incubated in medium containing 20 microM mirex. Lenses were removed from the medium at 2, 4, 6, and 8 h and assayed for K+ and Na+ concentrations and hydration. Mirex had no in vitro effect on these lens parameters compared to controls, and produced no changes in lens clarity. In contrast, incubation for 8 h in medium containing 20 microM kepone resulted in elevated lens Na+ and lower lens K+. Thus, cataracts induced by mirex in vivo are probably not due to a direct toxic effect on the lens.     

Induction of mixed-function oxidase activity in mouse lymphoid tissues by polycyclic aromatic hydrocarbons

Polycyclic aromatic hydrocarbon (PAH) exposure can cause mixed-function oxidase (MFO) enzyme induction in certain tissues of various organisms. Measurement of such induction might serve as a useful bioindicator of human exposure to PAHs, provided readily obtainable human tissues can be utilized for such measurements. We have investigated the MFO activity in various lymphoid tissues of the C3H mouse as a model system and have studied the effect of systemic PAH treatment on such enzyme activity. An MFO enzyme assay was used to measure the activity of 7-ethoxyresorufin deethylase, an enzyme activity that may be specific for the cytochrome P-448 subset of MFO enzymes (those enzymes that are induced in cells or tissues following PAH administration). Intraperitoneal injection of mice with 180 mg/kg (4.6 mg) benzo[a]pyrene (BaP) or 160 mg/kg (4.0 mg) 3-methylcholanthrene (MC) produced a significant induction in MFO activity in mouse spleen S9 fractions 48 h after the injection. Induction ratios (induced activity/control activity) between 4 and 5 were seen with BaP; MC produced induction ratios of 2.5-3.0. Enzyme activity was not induced in the spleen within 16 h following BaP or MC administration. Other experiments indicated that MFO activity could be induced in thymus cells 48 h after either BaP or MC treatment. Treatment with BaP or MC did produce significant enzyme induction in the liver and lung tissues from the animals both 16 and 48 h after chemical treatment.     

Effects of arsenite on hepatic mixed-function oxidase activity in rats

1. Injection of arsenite (As³⁺) to control rats results in losses of total hepatic cytochrome P-450 and significant decreases of ethoxycoumarin O-deethylase (ECOD) and ethoxyresorufin O-deethylase (EROD) activities. However, As³⁺ appears to decrease the activity of these enzymes differentially, with EROD showing greater sensitivity than ECOD.

2. Injection of As³⁺ to rats treated with phenobarbital and isosafrole significantly decreases the total content of hepatic cytochrome P-450 and various mixed function oxidase (MFO) activities, with the exception of ECOD which appears to be insensitive to As³⁺.

3. 3-Methylcholanthrene administration apparently protects against the effects of As³⁺ on the cytochrome P-450 system, since total content of the cytochrome P-450 and various MFO activities were all insensitive to this treatment.     

Induction and inhibition of hepatic drug metabolizing enzymes by rifampin.

The effects of the antibiotic rifampin on microsomal drug metabolizing enzymes were investigated. On acute administration, rifampin (100 mg/kg i.p.) doubled hexobarbital sleeping time and zoxazolamine paralysis time in mice. The in vitro metabolism of zoxazolamine (0.25 mM) and 17β-estradiol (10 μM) were inhibited 50% by rifampin in concentrations of 0.4 and 0.3 mM, respectively. The inhibition of ethylmorphine N-demethylase was competitive with an apparent K_i, of 52 μM. Repeated administration of rifampin to mice (50 mg/kg i.p. daily for 6 days) increased liver weight by 20%a cytochrome P-450 (50%), NADPH cytochrome c reductase (43%,). ethylmorphine N-demethylase (85%), zoxazolamine hydroxylase (77%), benzpyrene hydroxylase (174%). and 17β-estradiol metabolism (89%). Microsomal protein (mg/g). aniline hydroxylase and p-nitrophenol glucuronyl transferase activities were unaffected.Rat microsomal drug metabolizing enzyme activity was also inhibited after acute administration of rifampin as exemplified by an increase in hexobarbital sleeping time of 44% and a competitive inhibition of ethylmorphine N-demethylase by rifampin. The K_i, (33 μM) was close to that obtained with the mouse enzyme. The similarity in the in vivo and in vitro inhibition suggests that rifampin binds to microsomes in a similar manner in both species. Chronic administration of rifampin to rats (50 mg/kg i.p.) twice daily for six days did not lead to induction, indicating a species difference with respect to rifampin's inducing ability. Rifampin did not modify microsomal induction in rats by phenobarbital when both drugs were administered concomitantly. The mechanism responsible for the species difference and the clinical relevance of these results are discussed.     

Effects of acetone and methyl n-butyl ketone on hepatic mixed-function oxidase

The administration of ketones potentiates CCl4 hepatotoxicity; however, the potencies of the ketones differ. The aim of the present study was to assess potential differences between acetone and methyl n-butyl ketone (MnBK) on cytochrome P-450. The effects of single and repetitive doses of acetone and MnBK were determined in male rats by estimating the rate of metabolite formation of three substrates and the hepatic content of cytochrome P-450. A single treatment with acetone (13.5 mmol/kg or greater) enhanced the oxidation of aniline and 7-ethoxycoumarin, whereas repetitive treatments also increased aminopyrine demethylation and cytochrome P-450 content. Single and repetitive treatments of MnBK (15 mmol/kg) augmented the oxidation of all three substrates and increased cytochrome P-450 content. The effects of the ketones on cytochrome P-450 isozymes were characterized using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Acetone and MnBK increased the 52.1 and 54.1 kD forms and, in addition, MnBK tended to increase the 50.6 kD species. The data indicate that ketones differ in the type of isozymes induced and in the degree of induction. The higher potency of MnBK, compared to acetone, is probably associated with the fact that MnBK affects a greater number of isozymes than acetone.