Hepatic Microsomal Cytochrome P-450 Concentrations Research Articles

Effect of beta-naphthoflavone on o-tolyl saligenin phosphate-induced delayed neuropathy in two lines of chickens.

The effect of the microsomal enzyme inducer beta-naphthoflavone (beta NF) on the development of organophosphorus-induced delayed neuropathy (OPIDN) was examined in two laboratories (VPI and MSU), utilizing two strains of White Leghorn hens. A single intraperitoneal injection of beta NF at 80 mg/kg body weight 48 h prior to administration of o-tolyl saligenin phosphate (TSP), the neuroactive metabolite of tri-o-tolyl phosphate (TOTP), caused a significant increase in hepatic microsomal cytochrome P-450 concentrations and aniline hydroxylase activities after 72 h in both strains. Hepatic carboxylesterase and cholinesterase activities were not affected by beta NF treatment in either strain. Administration of TSP in single subcutaneous doses of 20 and 25 mg/kg body weight (VPI) or 30 and 60 mg/kg body weight (MSU) caused significant inhibition of whole-brain neuropathy target esterase (NTE) activity 24 h postdosing, and hens subsequently developed clinical signs characteristics of OPIDN. beta NF had no significant effect on NTE inhibition or on initiation or severity of OPIDN clinical signs. However, OPIDN clinical signs were less severe in the strain of bird (MSU) with the higher intrinsic hepatic carboxylesterase activity and the higher beta NF-induced cytochrome P-450 concentration. The study indicates that microsomal enzyme induction, which has been shown to alleviate TOTP-induced delayed neuropathy, could not alleviate OPIDN resulting from exposure to TSP. This study also suggests that strain may affect susceptibility to TSP-induced delayed neuropathy.

The influence of recombinant DNA-derived human and murine gamma interferons on mouse hepatic drug metabolism

Human gamma interferon given for up to 5 days by subcutaneous infusion or intraperitoneal injection did not significantly alter mouse hepatic microsomal oxidative drugmetabolizing enzyme activities. In contrast, murine gamma interferon and human alpha interferon given for 5 days at the same dose (10 7 units/kg) caused 25 and 50% decreases, respectively, in hepatic microsomal cytochrome P-450 concentrations. The human alpha interferon-induced decline in cytochrome P-450 was accompanied by a significant drop in p-nitroanisole demethylase activity and significant elevations in serum alanine aminotransferase and cytosolic glutathione S-transferase activities. An elevation in glutathione- S-transferase was the only significant change found following human gamma interferon administration. Microsomal UDP-glucuronosyltransferase activity was unaffected by any interferon.

Effects of pregnant mare serum gonadotrophin and oestradiol benzoate on enzymes associated with xenobiotic metabolism in immature and hypophysectomized female rats.

Treatment of immature (22-day-old) female Sprague-Dawley rats with pregnant mare serum gonadotrophin (PMSG) resulted in a significant increase in hepatic microsomal cytochrome P-450 concentrations when compared with animals treated with 0.9% (w/v) NaCl solution. Cytochrome P-450 concentrations in microsomes from control and PMSG-treated animals were 0.49 +/- 0.07 and 0.78 +/- 0.18 nmol/mg microsomal protein respectively (mean +/- S.D.). When hypophysectomized rats were treated with PMSG no significant difference was observed between saline-treated and PMSG-treated animals (0.84 +/- 0.06 vs 0.88 +/- 0.08 nmol/mg microsomal protein). Cytochrome b5 concentration and NADPH-cytochrome c reductase and ferricyanide reductase activities were not significantly affected by PMSG treatment. Similar results were obtained when immature and hypophysectomized rats were treated with oestradiol benzoate. Hepatic glutathione (GSH) S-transferase activity was significantly stimulated over control values when hypophysectomized or immature animals were treated with PMSG, oestradiol benzoate or 3-methylcholanthrene. Treatment of animals simultaneously with PMSG or oestradiol benzoate and 3-methylcholanthrene resulted in higher transferase activity than did either treatment alone. Oestradiol benzoate or PMSG treatment of immature female rats resulted in an approximate twofold increase in hepatic aryl hydrocarbon hydroxylase (AHH) activity. However, AHH activity in hypophysectomized animals was not significantly increased by PMSG or oestradiol benzoate treatment. It is concluded that PMSG or oestradiol benzoate treatment of immature female rats resulted in increased total cytochrome P-450 concentration and AHH activity in hepatic microsomes. Glutathione S-transferase activity was also stimulated. Neither PMSG nor oestradiol benzoate stimulated cytochrome P-450 levels or AHH activity in hypophysectomized rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Toxicological manifestations of 2,4,5,2',4',5'-, 2,3,6,2',3',6'-, and 3,4,5,3',4',5'-hexachlorobiphenyl and Aroclor 1254 in mink.

Adult female mink were fed diets that contained 2.5 ppm Aroclor 1254, 0.1 or 0.5 ppm 3,4,5,3',4',5'-hexachlorobiphenyl (345 HCB), 2.5 or 5.0 ppm 2,4,5,2',4',5'-hexachlorobiphenyl (245 HCB) or 2,3,6,2',3',6'-hexachlorobiphenyl (236 HCB), or a control diet from 1 mo prior to breeding through parturition. All mink fed 0.5 ppm 345 HCB died within 60 d, while those fed 0.1 ppm showed 50% mortality after 3 mo exposure. Only one stillborn kit was whelped in the Aroclor 1254 group. No adverse reproductive effects were observed in the animals fed 236 HCB or 245 HCB. Plasma progesterone concentrations were significantly depressed by Aroclor 1254 and significantly elevated by 0.1 ppm 345 HCB. 17 beta-Estradiol concentrations were not significantly altered by any of the dietary treatments. Hepatic microsomal cytochrome P-450 concentrations were significantly elevated by all treatments except 236 HCB, with the largest increases occurring in mink exposed to Aroclor 1254 and 345 HCB. Aminopyrine N-demethylase activity was elevated by 5.0 ppm 245 HCB. Aroclor 1254 caused significant elevations in both benzo[a]pyrene hydroxylase and ethoxyresorufin O-deethylase activities. Benzo[a]pyrene hydroxylase activities were also significantly elevated in mink fed 245 HCB and 345 HCB. Aroclor 1254 caused a significant elevation in cerebellar and hypothalamic norepinephrine concentrations and a significant elevation in hypothalamic dopamine concentrations. Cerebral dopamine was elevated by 0.1 ppm 345 HCB, while midbrain dopamine levels were depressed. Norepinephrine concentrations were significantly elevated by 5.0 ppm 236 HCB in the midbrain and by 5.0 ppm 245 HCB in the medulla.

Effect of Cholestasis Produced by Bile Duct Ligation on Hepatic Heme and Hemoprotein Metabolism in Rats

Cholestasis produced by bile duct ligation has been associated with decreased concentrations of hepatic microsomal cytochrome P450 and decreased hepatic microsomal oxidative drug metabolism. Bile duct ligation producing cholestasis results in a marked increase in hepatic microsomal heme oxygenase activity, with corresponding decreases in hepatic microsomal cytochrome P450 concentration, reduced form of nicotinamide adenine dinucleotide phosphate-cytochrome c reductase activity, and hepatic delta-aminolevulinic acid synthetase activity. As sham-operated rats also demonstrate a less prolonged decrease in cytochrome P450 concentration and reduced form of nicotinamide adenine dinucleotide phosphate-cytochrome c reductase activity, the metabolic effects of surgery and anesthesia must also be involved in these alterations in microsomal oxidative drug metabolism. The relative rate of hepatic cytochrome P450 synthesis and of degradation are both decreased after bile duct ligation. These data suggest that decreased hepatic microsomal cytochrome P450 concentrations in cholestasis are partly the result of decreased cytochrome P450 synthesis. Increased levels of heme oxygenase activity are not related to increased cytochrome P450 turnover, but may instead reflect enlargement and increased catabolism of a free heme pool resulting from decreased hemoprotein (cytochrome P450) synthesis.

Effect of hypoxia on carbon tetrachloride hepatotoxicity

The effect of hypoxia on carbon tetrachloride-induced hepatotoxicity was studied. Male rats were exposed to carbon tetrachloride for 2 hr in the presence of differing oxygen concentrations. Serum glutamate-pyruvate transaminase (SGPT) activities were measured 24 hr after the end of the exposure. Exposure of rats to 5000 ppm carbon tetrachloride in the presence of 100, 21,12, or 6% oxygen resulted in SGPT activities of 489, 420, 3768, and 1788 I.U./l respectively. Exposure of rats to air and 0, 1250, 2500, 5000, or 7500 ppm carbon tetrachloride gave SGPT activities of 35, 32, 69, 420, and 2188 I.U./l respectively; when 12% oxygen was used, the corresponding SGPT activities were 32, 665, 691, 3768, and 4200 I.U./l respectively. Exposure of rats to hypoxia produced histopathologically detectable condensation of hepatic cytoplasmic material, and exposure to 5000 ppm carbon tetrachloride in the presence of air produced mild centrilobular necrosis, which was much more severe when rats were exposed to 5000 pm carbon tetrachloride in the presence of 12% oxygen. Hepatic microsomal conjugated diene concentrations were increased by hypoxia and by exposure to carbon tetrachloride, but no synergistic interaction was observed. Hepatic microsomal cytochrome P-450 concentrations were decreased after exposure to carbon tetrachloride, but were the same after exposure to carbon tetrachloride and 12 or 21% oxygen. Hepatic carbon tetrachloride concentrations were the same in rats exposed to carbon tetrachloride in the presence of 12 or 21% oxygen; hepatic chloroform concentrations were higher in rats exposed to carbon tetrachloride in the presence of air than in the presence of 12% oxygen. The covalent binding of [ 14C]carbon tetrachloride metabolites to hepatic microsomal lipids and proteins was increased markedly by hypoxia as compared with normoxia. The covalent binding of metabolites of carbon tetrachloride to cellular macromolecules may play a role in the potentiation of carbon tetrachloride toxicity by hypoxia.

Carbon tetrachloride-induced changes in adrenal microsomal mixed-function oxidases and lipid peroxidation

Studies were carried out to compare the effects of carbon tetrachloride (CCl 4) in vivo and in vitro on adrenal and hepatic microsomal metabolism in guinea pigs. CCl 4 administration in vivo decreased adrenal and hepatic microsomal cytochrome P-450 concentrations and lowered benzphetamine (BZ) demethylase and benzo[ a]pyrene (BP) hydroxylase activities in both tissues. NADPH-cytochrome c reductase activity was decreased in hepatic but not in adrenal microsomes. Addition of CCl 4 to adrenal or hepatic microsomes in vitro produced a type I difference spectrum suggesting binding of CCl 4 to cytochrome(s) P-450; its magnitude was far greater in adrenal than in liver. Incubation of adrenal or hepatic microsomes in vitro with CCl 4 alone had little or no effect on mixed-function oxidase activity or on lipid peroxidation. However, when microsomes were incubated with CCl 4 + NADPH, the rates of BZ and BP metabolism were decreased, cytochrome P-450 concentrations were decreased, and lipid peroxidation was increased. The effects of CCl 4 + NADPH on enzyme activities were greater in adrenal than in hepatic microsomes. Addition of 1.0 m M EDTA or 0.1 m M MnCl 2 to the incubation medium blocked the effects of CCl 4 + NADPH on lipid peroxidation in adrenal and liver but had no effect on the decreases in mixed-function oxidase activities. The results indicate the following: (1) the adrenal cortex in the guinea pig is an active site of CCl 4 metabolism; (2) CCl 4 metabolism results in a loss of microsomal enzyme activities in the adrenal as well as liver; and (3) lipid peroxidation is not obligatory for the CCl 4-mediated destruction of microsomal enzymes.

Accelerated hepatic haem catabolism in the selenium-deficient rat.

1. Hepatic microsomal cytochrome P-450 concentrations are lower in selenium-deficient rats treated with phenobarbital for 4 days than in similarly treated control rats. 2. No defect in haem synthesis was found on the basis of measurements of delta-aminolaevulinate synthase (EC 2.3.1.37), delta-aminolaevulinate dehydratase (EC 4.2.1.24) and ferrochelatase (EC 4.99.1.1) activities, and urinary excretion of delta-aminolaevulinate, porphobilinogen, uroporphyrin and coproporphyrin. 3. No defect in apo-(cytochrome P-450) separated by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. 4. An increase in haem catabolism was found. An 8-fold increase in hepatic microsomal haem oxygenase (EC 1.14.99.3) activity occurred in selenium-deficient rats after phenobarbital treatment, compared with a less than 2-fold increase in control rats. Also excretion of 14CO in the breath after administration of delta-amino[5-14C]laevulinate was greater by phenobarbital-treated selenium-deficient rats than by similarly treated controls. 5. These studies demonstrate that the defective induction of cytochrome P-450 by phenobarbital in selenium-deficient rats is accompanied by increased haem catabolism. This could be due to increased breakdown of cytochrome P-450 or to catabolism of haem before it attaches to the apo-cytochrome. The role of selenium in stabilizing cytochrome P-450 and/or in protecting haem from breakdown remains to be determined.