Hepatic Cytochrome P-450 Content Research Articles

The Effect of Fever on Quinine and Quinidine Disposition in the Rat

The pharmacokinetics of quinine and its diastereoisomer quinidine has been investigated in normal and febrile rats. Endotoxin-induced fever in rats resulted in an increased quinine clearance (CL) (4.49 +/- 1.45 vs 1.38 +/- 0.65 L h-1 kg-1, P less than 0.001) and volume of distribution (Vd) (42.6 +/- 8.8 vs 28.9 +/- 10.3 L kg-1, P less than 0.05) with a concomitant shortening of the elimination half-life (t1/2) (7.1 +/- 2.5 vs 15.9 +/- 5.9 h, P less than 0.01). With quinidine, however, fever resulted in an increased CL (3.95 +/- 1.05 vs 1.89 +/- 0.60 L h-1 kg-1, P less than 0.002) with no change in Vd and a significant decrease in t1/2 (5.1 +/- 0.7 vs 10.1 +/- 2.8 h, P less than 0.001). In both studies there was no significant difference in hepatic microsomal protein or cytochrome P450 content. Neither drug accumulated in the liver but low concentrations of quinidine were present in the heart 24 h after administration. In-vitro studies suggest that temperature does not alter the binding of either drug. These data suggest that fever enhances the clearance of quinine and quinidine. These findings may offer some additional explanation of the lack of serious quinine and quinidine toxicity during the treatment of malaria infection, even after large dosages of the drug administered during the initial period of treatment when fever is most intense.

Sex-related differences in rat liver microsomal enzymes and their induction by doxapram.

The effects of doxapram on the hepatic microsomal mono-oxygenase system of male and female rats were investigated. Male and female rats were administered doxapram (10-120 mg kg-1 day-1, i.p.) for 4 days. In female rats, administration of doxapram (20, 40, 60, 80, 100 and 120 mg kg-1) elevated the parameters in a dose-dependent manner while doxapram (100 and 120 mg kg-1) elevated the levels of cytochrome P450 and hexobarbitone hydroxylase in male rats. Doxapram (40 mg kg-1) caused induction of hepatic drug metabolism typified by an increase of hepatic microsomal cytochrome P450 content and activities of hexobarbitone hydroxylase, benzphetamine N-demethylase and ethylmorphine N-demethylase in female rats, but no change in male rats. These findings were supported by the results of SDS/polyacrylamide-gel electrophoresis. However, 7-ethoxycoumarin O-de-ethylase and arylhydrocarbon hydroxylase activities were significantly increased in male rats. NADPH-cytochrome c reductase and NADH-cytochrome c reductase activities, and cytochrome b5 content were unaffected in rats of both sexes. The sex-dependent cytochrome P450 species may be selectively sensitive to the action of doxapram.

Impairment of drug metabolism by the liver in experimental fascioliasis in the rat.

Fascioliasis has been produced in the rat by an oral administration of 20 metacercariae of Fasciola hepatica. Hepatic microsomal cytochrome P450 and b5 contents and both aminopyrine demethylase and aniline hydroxylase activities have been measured during the course of the experimental distomiasis. The cytochrome P450 concentration and microsomal drug metabolizing enzymes generally fell by weeks 3 to 8 post-infestation and recovered to normal values thereafter. For the same period, both the histoenzymatically assayed liver cytochrome oxidase and arylsulphatase activities were reduced whereas there were correlated increases in glutamic pyruvic and glutamic oxaloacetic plasma transaminases. Tissue inflammation and destruction provoked by young histophagous migrating flukes could be responsible for these changes that have already been observed in several hepatic diseases. The possible influence of naturally-induced fasciolasis on liver drug metabolism is discussed.

Tissue Accumulation of Lanthanum as Compared to Aluminum in Rats with Chronic Renal Failure – Possible Harmful Effects after Long-Term Exposure

Background: Lanthanum (La) carbonate is a new treatment for hyperphosphatemia. We tested the effects of oral La carbonate and aluminum hydroxide, respectively, on tissue accumulation and liver function in rats with chronic renal failure (CRF). Methods: Adult male non-CRF and CRF rats were randomly assigned to 3 groups receiving either standard diet (St.D), or the same diet supplemented with 3% La carbonate (non-CRF La vs. CRF La) or 3% aluminum hydroxide (non-CRF Al vs. CRF Al). Results: After 12 weeks, serum phosphorus was decreased in both CRF La and Al groups. Urinary La and Al excretion was increased in these two groups, and so was liver and bone La content, and liver Al content. Both total body and liver weight were decreased in CRF La and CRF Al rats. Liver cell proliferation was decreased in these groups, while plasma total alkaline phosphatases and alanine aminotransferase were increased. Hepatic total cytochrome p450 content was reduced in CRF La, but not in CRF Al rats. Conclusion: Long-term oral La overload in rats with CRF was associated with a decrease in liver (and total body) weight and mild alterations of liver function, as was Al overload, possibly as a consequence of trace element accumulation.

Changes in the Gene Expression and Enzyme Activity of Hepatic Cytochrome P450 in Juvenile Sprague-Dawley Rats

The developmental changes in the hepatic cytochrome P450 (CYP) content, mRNA expression of 12 hepatic CYP subtypes, and the enzyme activities of 5 hepatic CYP subfamilies in rats were investigated using non-treated male and female Sprague-Dawley rats of ages postnatal day (PD) 4, 16, 30 and 8 and 12 weeks. The hepatic proliferation kinetics was also determined by using the phospho-histone H3 (p-histon)-labeled hepatocyte index. The developmental changes in the enzyme activities of hepatic expression of CYP1A and CYP3A in rats were similar to those in humans, although there is no fetal-neonatal dominant CYP3A subtype in rat livers unlike human CYP3A7. On the other hand, the developmental pattern of expression of the CYP2C subfamily differed between humans and rats. Enzyme activity and mRNA expression of each hepatic CYP subtype in rats on PD 30 was similar to that after 8 weeks of age, except in the case of sex-dependent CYP subtypes. The p-histon-labeled hepatocyte index was approximately 10-fold higher in PD 30 rats than in 8-week-old rats. Therefore, the livers of juvenile rats, which have high hepatocellular proliferation activity and a sufficient amount of metabolic enzymes such as CYP, may be more sensitive to the cytotoxic and carcinogenic effects of chemicals than the livers of adult rats. Thus, our results on developmental difference of hepatic CYPs in juvenile rats are useful to identify underlying age-dependent susceptibility of chemical-induced toxicity, and to understand developmental change of chemical disposition.

Tertiary butyl alcohol in drinking water induces phase I and II liver enzymes with consequent effects on thyroid hormone homeostasis in the B6C3F1 female mouse

Tertiary butyl alcohol (TBA) was administered to groups of 15 female B6C3F1 mice in drinking water at concentrations of 0, 2.0 or 20 mg TBA ml(-1), for 14 days, for assessment of gross and histological changes in the liver and thyroid, thyroid hormones (T3, T4, and TSH), total hepatic cytochrome P450 (Cyp) content, specific Cyp activities and quantitative PCR analysis of specific Cyp enzymes (Cyp1a1, Cyp2b9, Cyp2b10, Cyp3a11), sulfuryltransferases (ST1a1, ST2a2, and STn) and glucuronyltransferases (UGT1a1, UGT2b1, and UGT2b5). Phenobarbital (PB) was administered to a positive control group by oral gavage at a daily dose of 80 mg kg(-1). TBA caused, on day 14, a reduction in circulating T3 (12-15% decrease) and a dose-dependent reduction in T4 (13-22% decrease), with no evidence of thyroid pathology. Two of five livers examined in the 20 mg TBA ml(-1) dose group showed mild, diffuse centrilobular hypertrophy. On day 14, Cyp 7-benzoxyresorufin-O-debenzylase activity was significantly induced 12-fold by TBA at 20 mg ml(-1), and 1.8-fold at the 2.0 mg TBA ml(-1) concentration. Cyp 7-pentoxyresorufin-O-dealkylase activity was slightly induced (2.1-fold) by 20 mg TBA ml(-1) on day 14. Quantitative PCR analysis of gene transcripts showed a significant induction of Cyp2b10 and ST1a1 with both TBA concentrations, and a slight induction of Cyp2b9 at 20 mg TBA ml(-1) only. PB induced all phase I and phase II gene transcripts except for Cyp1a1 and Cyp2b9. These findings suggest that TBA, at and below doses used in chronic studies, is an inducer of phase I and phase II liver enzymes, with resulting decreases in circulating thyroid hormones in B6C3F1 mice.

The Effect of Infliximab on Hepatic Cytochrome P450 and Pharmacokinetics of Verapamil in Rats with Pre-Adjuvant Arthritis: A Drug-Disease and Drug-Drug Interaction

Inflammatory conditions result in increased concentration but reduced potency of some cardiovascular drugs. This is associated with increased levels of pro-inflammatory mediators. Infliximab reduces pro-inflammatory mediators and reverses the diminishing effect of inflammation on response in the rat. We suggested that infliximab treatment would also reverse the effects of inflammation on drug metabolism and clearance. We examined hepatic cytochrome P450 content and the pharmacokinetics of verapamil in pre-adjuvant arthritic rats treated with infliximab. Pre-adjuvant arthritis was induced in male Sprague-Dawley rats with a tail base injection of Mycobacterium butyricum. Animals were monitored for symptoms of arthritis, serum nitrite and C-reactive protein. On day 6, rats were administered with single s.c. doses of infliximab (10 mg/kg). On day 14, a single i.v. dose of racemic verapamil (2 mg/kg) was administered, and S- and R-verapamil concentrations were determined by high performance liquid chromatography. Hepatic cytochrome P450 content and verapamil protein binding were also measured. Serum nitrite levels were significantly elevated in pre-adjuvant arthritis. Infliximab did not affect mean nitrite concentrations but there was a significant correlation between nitrite and S-verapamil concentrations as well as cytochrome P450, CYP3A, and CYP1A contents. Infliximab increased cytochrome P450 enzymes content that had been diminished by pre-adjuvant arthritis but had no significant effect on verapamil protein binding. Infliximab partially restores hepatic cytochrome P450 enzyme contents. The effect of infliximab on the mean verapamil clearance was not significantly affected due, likely, to the lack of effect on plasma protein binding.

Mode of Action Analysis for the Synthetic Pyrethroid Metofluthrin-Induced Rat Liver Tumors: Evidence for Hepatic CYP2B Induction and Hepatocyte Proliferation

Two-year treatment with high doses of Metofluthrin produced hepatocellular tumors in both sexes of Wistar rats. To understand the mode of action (MOA) by which the tumors are produced, a series of studies examined the effects of Metofluthrin on hepatic microsomal cytochrome P450 (CYP) content, hepatocellular proliferation, hepatic gap junctional intercellular communication (GJIC), oxidative stress and apoptosis was conducted after one or two weeks of treatment. The global gene expression profile indicated that most genes with upregulated expression with Metofluthrin were metabolic enzymes that were also upregulated with phenobarbital. Metofluthrin induced CYP2B and increased liver weights associated with centrilobular hepatocyte hypertrophy (increased smooth endoplasmic reticulum [SER]), and induction of increased hepatocellular DNA replication. CYP2B1 mRNA induction by Metofluthrin was not observed in CAR knockdown rat hepatocytes using the RNA interference technique, demonstrating that Metofluthrin induces CYP2B1 through CAR activation. Metofluthrin also suppressed hepatic GJIC and induced oxidative stress and increased antioxidant enzymes, but showed no alteration in apoptosis. The above parameters related to the key events in Metofluthrin-induced liver tumors were observed at or below tumorigenic dose levels. All of these effects were reversible upon cessation of treatment. Metofluthrin did not cause cytotoxicity or peroxisome proliferation. Thus, it is highly likely that the MOA for Metofluthrin-induced liver tumors in rats is through CYP induction and increased hepatocyte proliferation, similar to that seen for phenobarbital. Based on analysis with the International Life Sciences Institute/Risk Science Institute MOA framework, it is reasonable to conclude that Metofluthrin will not have any hepatocarcinogenic activity in humans, at least at expected levels of exposure.

Phenobarbital (PB)-induced changes in blood coagulationrelated parameters in pregnant rats, lactating rats and pups

Effects of repeated administration of phenobarbital (PB) on blood coagulation-related parameters were examined in non-pregnant, pregnant and lactating rats, and also in pups born to PB-treated lactating dams. PB was orally administered at a dose level of 80 mg/kg/day to pregnant (from gestation day (GD) 13), postpartum (from postpartum day (PPD) 7) and non-pregnant rats (from 13 weeks of age) for 7 days. Blood was collected on GD20 or PPD14 to perform blood coagulation examination. Concurrently, the blood coagulation parameters were examined in the pups. Increases in liver weight and/or hepatic cytochrome P450 content were observed in the PB-treated non-pregnant, pregnant and lactating rats. Activated partial thromboplastin time (APTT) was prolonged and anti-thrombin III (ATIII) concentration was increased in the lactating rats, while there were no changes in prothrombin time (PT) or APTT in the non-pregnant and pregnant rats. Moreover, prolongation of PT and APTT and decreases in factors VII and IX activities were observed in their pups. Thus, prolongation of blood coagulation time was confirmed in both dams and their pups following PB-administration to lactating dams. Effects of vitamin K(2) (VK(2)) on PB-induced changes in blood coagulation-related parameters of both dams and their pups were examined by co-administration with PB and VK(2) to lactating dams. PT and APTT were comparable to the control and PB-induced prolongation of blood coagulation time was improved in the pups while APTT was prolonged in dams, suggesting that VK(2) was beneficial to pups but not to dams.

Changes of midazolam pharmacokinetics in Wistar rats treated with lipopolysaccharide: relationship between total CYP and CYP3A2

It has been reported that infection interferes with drug metabolism, resulting in changes in pharmacokinetics. In this study, we investigated the effects of lipopolysaccharide (LPS) on hepatic total cytochrome P450 (CYP), CYP3A2, and CYP2C11 contents in a transient, LPS-induced, endotoxemia model of rats. In addition, to assess the effects on CYP3A2 activities, the pharmacokinetics of midazolam (CYP3A2 substrate) and 1-OH-midazolam (metabolite of midazolam) were investigated. Hepatic total CYP contents were significantly low until day 3 (P < 0.05) but returned to the control level on day 5. Hepatic CYP3A2 contents were significantly decreased on day 1 until day 5 (P < 0.05) but returned to the control level on day 7. Hepatic CYP2C11 contents were continuously low until day 7, and lowest on day 3. The AUC of 1-OH-midazolam was significantly decreased on day 1 after LPS administration (P < 0.01). In conclusion, LPS (5 mg/kg) challenge decreased hepatic total CYP, CYP3A2, and CYP2C11 contents and also decreased the activities of hepatic CYP3A2. It took at least 7 days for hepatic total CYP and CYP3A2 to recover to control levels, and it was suggested that the changes of hepatic total CYP contents might correlate with those of hepatic CYP3A2 contents and activities. Additionally, it is shown that their changes might reflect the recovery process from inflammation.

CYP2B6: New Insights into a Historically Overlooked Cytochrome P450 Isozyme

Human CYP2B6 has been thought to account for a minor portion (<1%) of total hepatic cytochrome P450 (CYP) content and to have a minor function in human drug metabolism. Recent studies, however, indicate that the average relative contribution of CYP2B6 to total hepatic CYP content ranges from 2% to 10%. An increased interest in CYP2B6 research has been stimulated by the identification of an ever-increasing substrate list for this enzyme, polymorphic and ethnic variations in expression levels, and evidence for cross-regulation with CYP3A4, UGT1A1 and several hepatic drug transporters by the nuclear receptors pregnane X receptor and constitutive androstane receptor. Moreover, 20- to 250-fold interindividual variation in CYP2B6 expression has been demonstrated, presumably due to transcriptional regulation and polymorphisms. These individual differences may result in variable systemic exposure to drugs metabolized by CYP2B6, including the antineoplastics cyclophosphamide and ifosfamide, the antiretrovirals nevirapine and efavirenz, the anesthetics propofol and ketamine, the synthetic opioid methadone, and the anti-Parkinsonian selegiline. The potential clinical significance of CYP2B6 further enforces the need for a comprehensive review of this xenobiotic metabolizing enzyme. This communication summarizes recent advances in our understanding of this traditionally neglected enzyme and provides an overall picture of CYP2B6 with respect to expression, localization, substrate-specificity, inhibition, regulation, polymorphisms and clinical significance. Emphasis is given to nuclear receptor mediated transcriptional regulation, genetic polymorphisms, and their clinical significance.

Antihyperalgesic effect of a Cannabis sativa extract in a rat model of neuropathic pain: mechanisms involved

This study aimed to give a rationale for the employment of phytocannabinoid formulations to treat neuropathic pain. It was found that a controlled cannabis extract, containing multiple cannabinoids, in a defined ratio, and other non-cannabinoid fractions (terpenes and flavonoids) provided better antinociceptive efficacy than the single cannabinoid given alone, when tested in a rat model of neuropathic pain. The results also demonstrated that such an antihyperalgesic effect did not involve the cannabinoid CB1 and CB2 receptors, whereas it was mediated by vanilloid receptors TRPV1. The non-psychoactive compound, cannabidiol, is the only component present at a high level in the extract able to bind to this receptor: thus cannabidiol was the drug responsible for the antinociceptive behaviour observed. In addition, the results showed that after chronic oral treatment with cannabis extract the hepatic total content of cytochrome P450 was strongly inhibited as well as the intestinal P-glycoprotein activity. It is suggested that the inhibition of hepatic metabolism determined an increased bioavailability of cannabidiol resulting in a greater effect. However, in the light of the well known antioxidant and antiinflammatory properties of terpenes and flavonoids which could significantly contribute to the therapeutic effects, it cannot be excluded that the synergism observed might be achieved also in the absence of the cytochrome P450 inhibition.

Blood coagulation-related parameter changes in Sprague-Dawley (SD) rats treated with phenobarbital (PB) and PB plus vitamin K

Effects of dose and duration of phenobarbital (PB) administration and those of co-administration of PB and vitamin K on blood coagulation-related parameters were examined in specific pathogen-free (SPF) rats of Sprague-Dawley strain kept on an ordinary diet. In Experiment 1, oral administration of PB (0, 25, 50, 100 or 150 mg/kg/day) for 2 weeks induced increases in hepatic cytochrome P450 content and CYP2B expression, prolongation of coagulation time (activated partial thromboplastin time (APTT) and Thrombotest (TBT)) and an increase in anti-thrombin III (AT III) concentration in a dose-dependent manner. In Experiment 2, PB administration (100 mg/kg/day) for up to 14 days produced time-dependent increases in hepatic cytochrome P450 content and CYP2B (CYP2B1 and CYP2B2) expression. APTT was prolonged from day 1 and AT III concentration was increased from day 2, whereas the coagulation time (TBT) was prolonged from day 7. In Experiment 3, APTT prolonged by PB (100 mg/kg/day) was shortened after vitamin K(2) (30 mg/kg/day) co-administration, although AT III concentration was still increased. This suggests that not AT III but PB-induced vitamin K deficiency may play an important role in PB-induced prolongation of coagulation time in SPF rats kept on an ordinary diet.

Pomegranate Juice Effects on Cytochrome P450s Expression:In VivoStudies

Beneficial health effects have recently been claimed for pomegranate juice. In vitro and in vivo studies have demonstrated its anti-atherosclerotic capacity, chemoprevention and chemotherapy of prostate cancer, and antiproliferative, apoptotic, and antioxidant activity, among others. On the other hand, there is a complex interplay between tumor initiation, promotion, and progression and xenobiotic biotranformation. This led us to investigate the effect of pomegranate juice consumption on cytochrome P450 (CYP) activity and expression. For this purpose, male mice consumed this fruit juice for 4 weeks, and pentobarbital-induced sleeping time and total hepatic CYP content, activity, and expression were evaluated. Moreover, the activity of CYP isoform 2E1 and expression of the main CYP isoforms, namely, CYP1A1/2, CYP2E1, and CYP3A, were also assessed. It was found that pomegranate juice consumption decreased total hepatic CYP content as well as the expression of CYP1A2 and CYP3A. Prevention of procarcinogen activation through CYP activity/expression inhibition may be involved in pomegranate juice's effect on tumor initiation, promotion, and progression.

N-nitrosodimethylamine induced changes in the activities of carcinogen-metabolizing enzymes in the liver of male mice: role of glutathione and gossypol as antioxidants

The importance of nutrition in protecting living organisms from the toxic effects of environmental carcinogens has attracted the attention of many researchers. In order to exert their toxic and/or carcinogenic effects, most carcinogens need to be activated primarily by phase I drug-metabolizing enzymes including; cytochrome P450, cytochrome b5, arylhydrocarbon hydroxylase (AHH), N-nitroso-dimethylamine N-demethylase I (NDMA-N-dI), NADPH-cytochrome c reductase and the expression of cytochrome P450 2E1. Changes in the activity of these enzymes were determined in livers of male mice pretreated with either glutathione (GSH) or gossypol as antioxidants, for seven consecutive days before administration of NDMNA as single dose for two hours before decapitation of mice. The total hepatic content of cytochrome P450 and AHH activity were induced, whereas the activity of NDMA-N-dI and expression of cytochrome P450 2E1 were reduced after treatment of mice with NDMA as a single dose. On the other hand, preteatments of mice with either gossypol or GSH for seven consecutive days decreased the total cytochrome P450 content as well as AHH activity whereas the activity of NDMA-N-dI and the expression of cytochrome P450 2E1 increased. Treatment of mice with NDMA only induced the total hepatic content of cytochrome P450 and AHH activity. Interestingly pretreatment of mice with GSH for seven consecutive days before injection of NDMA was found to restore the induced cytochrome P450 and AHH activity caused by NDMA to their normal levels. However, pretreatment of mice with gossypol prior to administration of NDMA did not restore such activities to their normal levels. Treatment of mice with either GSH or gossypol induced the expression of cytochrome P450 2E1. Interestingly, gossypol/NDMA treated mice restored the induced cytochrome P450 2E1 expression, caused by gossypol, to its normal level. However, GSH/NDMA-treated mice did not restore the induced level of cytochrome P450 2E1 caused by GSH, to its normal level. It is concluded that, pretreatment of mice with GSH and gossypol prior to administration of NDMA induced the expression of cytochrome P450 2E1. Therefore, GSH and gossypol, could induce the toxicity and/or carcinogenicity of N-nitrosamines whereas they might protect the liver and probably other organs from the toxic and carcinogenic effects of other carcinogens such as polycyclic aromatic hydrocarbons, e.g. benzo[a]pyrene, through inhibition of total cytochrome P450 content and AHH activity. Key words: Aryl hydrocarbon hydroxylase, Cytochrome P450 2E1, N-nitrosodimethylamine N- demethylase, glutathione, antioxidants.

A mode of action for induction of liver tumors by Pyrethrins in the rat

High doses of Pyrethrins produce liver tumors in female rats. To elucidate the mode of action for tumor formation, the hepatic effects of Pyrethrins have been investigated. Male Sprague-Dawley CD rats were fed diets containing 0 (control) and 8000 ppm Pyrethrins and female rats' diets containing 0, 100, 3000 and 8000 ppm Pyrethrins for periods of 7, 14 and 42 days and 42 days followed by 42 days of reversal. As a positive control, rats were also fed diets containing 1200-1558 ppm sodium Phenobarbital (NaPB) for 7 and 14 days. The treatment of male rats with 8000 ppm Pyrethrins, female rats with 3000 and 8000 ppm Pyrethrins and both sexes with NaPB resulted in increased liver weights, which were associated with hepatocyte hypertrophy. Hepatocyte replicative DNA synthesis was also increased by treatment with Pyrethrins and NaPB. The treatment of male and female rats with Pyrethrins and NaPB produced significant increases in hepatic microsomal cytochrome P450 (CYP) content and a marked induction of CYP2B-dependent 7-pentoxyresorufin O-depentylase and testosterone 16beta-hydroxylase activities. Significant increases were also observed in CYP3A-dependent testosterone 6beta-hydroxylase activity. The hepatic effects of Pyrethrins were dose-dependent in female rats with 100 ppm being a no effect level and on cessation of treatment were reversible in both sexes. This study demonstrates that Pyrethrins are mitogenic CYP2B form inducers in rat liver. The mode of action for Pyrethrins-induced rat liver tumor formation appears to be similar to that of NaPB and some other non-genotoxic CYP2B inducers of hepatic xenobiotic metabolism.

Effects of exposure to the water‐soluble fraction of crude oil on the swimming performance and the metabolic and ionic recovery postexercise in Pacific herring (Clupea pallasi)

The swimming performance and recovery from exercise were determined in juvenile Pacific herring (Clupea pallasi) following exposure to the water-soluble fraction (WSF) of North Slope crude oil for more than eight weeks. Total polycyclic aromatic hydrocarbon concentrations (mean +/- standard error) at the beginning of exposures were as follows: control, 0.2 +/- 0.1 microg/L; low, 9.6 +/- 2.5 microg/L; medium, 40.7 +/- 6.9 microg/L; and high, 120.2 +/- 11.4 microg/L. Biological availability of hydrocarbons was confirmed by a significant induction of hepatic cytochrome P450 content and ethoxyresorufin-O-deethylase activity. Critical swimming speed (Ucrit) was significantly reduced in fish exposed to the highest concentration of WSF for 96 h (11% +/- 3.7% reduction) and at the two highest concentrations at four weeks (16% +/- 3.6% and 29% +/- 5.4% reductions) and eight weeks (11% +/- 3.8% and 40% +/- 5.7% reductions). Mortality occurred in all groups 24 h following Ucrit swim trials, with significantly higher mortalities observed in fish exposed to WSF in a concentration- and time-dependent manner (maximum mortality of 72.2% +/- 5.5% in the eight-week, high-exposure group). Burst swimming alone resulted in increased plasma cortisol, lactate, Na+, and Cl- concentrations and decreased muscle glycogen levels that returned to baseline values by 24 h. An interpretation of the effect of WSF exposure on postexercise metabolic recovery was complicated by pre-exercise alterations in several parameters. The time courses and magnitudes of several key postexercise parameters, including plasma cortisol, lactate, and muscle glycogen, were significantly altered by exposure to WSE The present study clearly shows that hydrocarbon exposure can reduce the swimming ability of fish and their ability to recover from exhaustive exercise.

Modulation of antipyrine clearance by polysaccharide peptide (PSP) isolated from Coriolus versicolor in the rat

Polysaccharide peptide (PSP), isolated from Coriolus versicolor COV-1, has been previously shown to have immuno-stimulatory, anti-tumour and analgesic effects in animal models. When used as an adjunct in cancer chemotherapy in clinical trials carried out in China, PSP improved the quality of life in the patients by improving appetite and alleviating symptoms associated with cancer chemotherapy. In this study, the effects of non-toxic doses of PSP on phase I metabolism was investigated in the rat, using the conventional probe antipyrine. Acute PSP (3–5 μmol/kg, i.p.) treatment did not produce significant changes in antipyrine clearance. Sub-chronic treatment with PSP (1–3 μmol/kg/day, i.p., 3 days) decreased the antipyrine clearance (30–35%), with an increase in the plasma half-life ( T 1/2) by 55% and an increase in the area under concentration–time curve (AUC) by 61%. Total hepatic cytochrome P450 (P450) was dose-dependently decreased (32–54%) after sub-chronic, but not the acute treatment of PSP. Given that PSP can affect phase I metabolism and hepatic cytochrome P450 content, the concomitant use of PSP with other therapeutic agents that undergo phase I metabolism should be carefully monitored.

Glutathione Alleviates the Influence of N-Nitrosamines on the Activity of Carcinogen-Metabolizing Enzymes in the Liver of Male Mice

Most xenobiotics and carcinogens are metabolized primarily by the hepatic drug-metabolizing enzymes including cytochrome P450, cytochrome b5, arylhydrocarbon hydroxylase (AHH), and dimethylnitrosamine N-demethylase I (DMN-dI) as phase I drug-metabolizing enzymes. Therefore, changes in the activities of the above mentioned enzymes as well as free radical levels measured as thiobarbituric acid reactive substances [TBARS] were determined in the livers of sixty mice pretreated with glutathione [100 mg/kg], as antioxidant, before intraperitoneal administration of dimethylnitrosamine [DMN] [10 mg/kg], diethylnitosamine [DEN] [10 mg/kg], or methylpropylnitrosamine [MPN] [10 mg/kg] for seven consecutive days. Ten control animals received distilled water as vehicle for comparison with treated animals. The hepatic content of cytochrome P450 induced after treatment of mice with DMN, DEN and MPN while DMN, DEN, or MPN increased AHH activity. Also, cytochrome b5 content and DMN-N-dI induced after treatment with DMN or DEN. Interestingly, glutathione pretreatment restored the activities of the previously mentioned enzymes caused by DMN or DEN. On the other hand, DMN, DEN, and MPN depleted the levels of glutathione, and increased the levels of free radicals [TBARS]. DMN and DEN increased the activity of GST. Interestingly, pretreatment of mice with glutathione before administration of DMN, or DEN was found to restore the levels of TBARS and GSH to their normal levels. In addition, GSH pretreatments induced the activity of GST above the normal levels after administration of DMN or MPN except DEN did not change such activity. It was concluded that N-nitrosamines enhanced the activity of phase I & II carcinogen-metabolizing enzymes as well as increased the production of free radicals. The mechanism by which GSH restores the activity of carcinogen-metabolizing enzymes or free radical production to their normal levels is possibly due to induction of GST activity and/or a direct binding to the xenobiotic molecules.

Selective protection of curcumin against carbon tetrachloride-induced inactivation of hepatic cytochrome P450 isozymes in rats

We investigated the effects of curcumin, a major antioxidant constituent of turmeric, on hepatic cytochrome P450 (CYP) activity in rats. Wistar rats received curcumin-containing diets (0.05, 0.5 and 5 g/kg diet) with or without injection of carbon tetrachloride (CCl 4). The hepatic CYP content and activities of six CYP isozymes remained unchanged by curcumin treatment, except for the group treated with the extremely high dose (5 g/kg). This suggested that daily dose of curcumin does not cause CYP-mediated interaction with co-administered drugs. Chronic CCl 4 injection drastically decreased CYP activity, especially CYP2E1 activity, which is involved in the bioactivation of CCl 4, thereby producing reactive free radicals. Treatment with curcumin at 0.5 g/kg alleviated the CCl 4-induced inactivation of CYPs 1A, 2B, 2C and 3A isozymes, except for CYP2E1. The lack of effect of curcumin on CYP2E1 damage might be related to suicidal radical production by CYP2E1 on the same enzyme. It is speculated that curcumin inhibited CCl 4-induced secondary hepatic CYPs damage through its antioxidant properties. Our results demonstrated that CYP isozyme inactivation in rat liver caused by CCl 4 was inhibited by curcumin. Dietary intake of curcumin may protect against CCl 4-induced hepatic CYP inactivation via its antioxidant properties, without inducing hepatic CYPs.