Olive oil protects rat liver microsomes against benzo(a)pyrene-induced oxidative damages: an in vitro study.

Benzo(a)pyrene (B(a)P), a member of the polycyclic aromatic hydrocarbon family is present ubiquitously in the environment. One of its toxic effects is induction of oxidative stress (mediated by the enzyme B(a)P hydroxylase) which leads to various diseases like cancer. Olive oil (OO) that consists of many antioxidant compounds is reported to have many beneficial properties including protection against cancer. The objective of the present study is to evaluate the effect of OO on B(a)P hydroxylase enzyme and further elucidate the antioxidant capacity of OO against B(a)P-induced toxicity. Rat liver microsomes were divided into three groups: vehicle control, B(a)P treated group, and OO + B(a)P co-incubated group. Antioxidant enzymes which were decreased and protein carbonyl content and lipid peroxidation products which were increased on exposure to B(a)P was attenuated to near normal on OO exposure. B(a)P hydroxylase enzyme was very low in OO incubated group which may be due to inhibition of the enzyme by OO or high utilization for the metabolism of B(a)P. Further, no B(a)P metabolites (3-OH B(a)P and B(a)P 7,8-dihydrodiol) were identified in HPLC during B(a)P + OO exposure. The results prove the protective role of OO against B(a)P-induced oxidative damage.

[Effect of soybean isoflavones on antioxidant status in rats fed diet with oxidized linseed oil]. / Vliianie izoflavonov soi na antioksidantnyi status krys, poluchavshikh ratsion s okislennym l'nianym maslom.

The maintenance of male Wistar rats on semi-synthetic diets containing oxidized flaxseed oil for 4 weeks caused decrease in serum total antioxidant activity, significant suppression of microsomal UDP-glucuronosyl transferase activity and benzpyrene hydroxylase activity, increase in reduced glutathione level and catalase activity of liver. Supplementation of diets with 0.05% of soy isoflavones has resulted in normalization of investigated characteristics.

Species differences in hepatic pulmonary and upper gastrointestinal tract biotransformation enzymes on long-term feeding of masheri--a pyrolyzed tobacco product.

The activities of several activating enzymes and that of glutathione S-transferase as well as levels of glutathione were measured in the upper alimentary tract, lung, and liver of Swiss mice, Sprague-Dawley rats, and Syrian golden hamsters treated with 10% masheri (pyrolyzed tobacco) in diet for 20 months. Significant increase in activities of phase I activating enzymes and a remarkable decrease in the phase II detoxification system in most extrahepatic tissues of the treated animals of all three species was observed. These observations suggest that the prolonged exposure to environmental xenobiotics/carcinogens affects the drug-metabolizing enzymes of the gastrointestinal tract, which may be an important factor in determining the susceptibility of different organs to carcinogen exposure.

Carcinogenicity studies of masheri: pyrolysed tobacco product, in vitamin-A-deficient Sprague Dawley rats.

The carcinogenicity of long-term feeding of masheri extract to animals in a vitamin-A-sufficient (SLO+) and deficient (SLO-) state was studied in Sprague Dawley rats by feeding daily dose of 3 mg extract over a period of 21 months. The phase I activating enzymes, the glutathione (GSH)/glutathione S-transferase (GST) detoxification system, and the hepatic and circulating levels of vitamins A and C were also monitored at 12 and 21 months. It was observed that the phase I enzyme activities were significantly higher in SLO+ than in SLO- rats at both 12 months and 21 months. Moreover, the SLO- masheri-treated animals also showed a decreased in the GSH/GST detoxification system while the reverse was observed in SLO+ group. Masheri extract treatment significantly lowered the hepatic and circulating levels of vitamin A while a concurrent increase was observed in the vitamin C level. The extract was found to be tumorigenic in both the SLO+ and SLO- groups. Benign tumours were observed in the SLO+ group while a high incidence of malignant tumours of the lung were observed in the SLO- group upon treatment with masheri extract.

[Evaluation of toxic effects on yusho causal substances by chick embryo hepatic microsomal enzymes activities].

PCBs, non-ortho chlorine substituted PCBs (Co-PCBs), PCQs and (PCDFs + PCDDs), all of which contained similar compositions of those corresponding in yusho oil, were prepared from a PCB preparation used as a heat exchanger fluid. After dissolved in 1, 4-dioxane, they were applied into the air sac of white leghorn eggs incubated for 16.5 days at 37.5 degrees C. Forty eight hours after injection, the hepatic benzo(a)pyrene hydroxylase (AHH) and 7-ethoxyresorufin deethylase (EROD) activities were assayed. The average relative potencies of induction for the two microsomal drug metabolizing enzymes by (PCDFs + PCDDs), Co-PCBs, PCBs and PCQs were 100, 13.4, 0.0006 and 0.0004, respectively. The toxic effects for yusho disease by these substances were calculated from the relative enzyme induction potencies and the average concentrations in yusho oils with the production dates of February 9 and 10, 1968. Consequently, the relative toxicities of (PCDFs+PCDDs), Co-PCBs, PCBs and PCQs were 100, 13.2, 0.06 and 0.12, respectively. This result, as well as our previous investigations using rats and monkeys, insists that (PCDFs+PCDDs) are the primary causal agents for yusho disease. However, the Co-PCBs, which were recently detected in the yusho oils by us, were revealed to be fairly effective in yusho manifestation. In addition, it was cleared that the hepatic enzyme induction by the Co-PCBs fraction, which contained other several PCB isomers, was almost completely contributed by only Co-PCBs such as 3,4,3',4'-tetra- 3,4,5,3',4'-penta- and 3,4,5,3',4',5'-hexachlorinated biphenyls present in the fraction. A chemical uptake rate from the air sac by the chick embryo decreased significantly in the cases of extremely high doses of PCBs (10,000 micrograms/egg) and PCQs (3,333 and 10,000 micrograms/egg), and result the elevations of hepatic enzymes activities were depressed, indicating that the suitable chemical dose amount to be less than about 1,000 micrograms/egg.

The role of heme metabolism during the induction of hepatic and renal cytochrome P-450 levels and drug-metabolizing enzymes in rats by a Prudhoe Bay crude oil.

Administration of Prudhoe Bay crude oil (PBCO) to rats resulted in a dose-related increase in liver weight; rapid and marked increase in the activity of hepatic delta-aminolevulinate synthetase, the initial and rate-limiting enzyme in the heme biosynthetic pathway; rapid decline in the activity of hepatic heme oxygenase, the rate-limiting enzyme of heme catabolism; and more gradual increase in the levels of hepatic cytochrome P-450 and some mixed-function oxidase activities such as benzo[a]pyrene hydroxylase and 7-ethoxyresorufin-O-deethylase. PBCO treatment also increased renal cytochrome P-450 levels and mixed-function oxidase activities; however, delta-aminolevulinate synthetase and heme oxygenase activities were unchanged. This suggests that different regulatory mechanism(s) may be involved in renal heme metabolism and induction of monoxygenase system.

Alterations in platelet aggregation and microsomal benzo-alpha-pyrene hydroxylase activities after exposure of rats to a Prudhoe Bay crude oil.

Administration of a Prudhoe Bay crude oil (PBCO) to rats has been shown to (a) inhibit platelet aggregation induced by adenosine diphosphate (ADP), arachidonic acid, or epinephrine and (b) induce benzo-alpha-pyrene hydroxylase (BPH) in the liver and small intestine. Maximum inhibition of aggregation (90%) was seen 12 to 16 hours subsequent to dosing. However, substantial inhibition was observed as early as four hours and as late as 48 hours after dosing. Of particular interest was the sensitivity of the platelet response compared with the putatively sensitive response of monooxygenase induction in liver. As little as 0.1 ml of PBCO per kilogram body weight (bw) caused an inhibition of aggregation with all three agonists. A similar inhibition of the release of ADP from platelets in the presence of arachidonic acid or epinephrine was also observed. In contrast, hepatic BPH activity showed only a modest increase (67%) over the control value even after administration of 2 ml of PBCO per kilogram body weight. Small intestine BPH activity was more sensitive, showing a gradual increase of up to 19-fold 24 hours after dosing with 2 ml of PBCO per kilogram body weight. The sensitivity of the platelet response is of general environmental interest and evaluating platelet aggregation in humans may be important as a noninvasive assay for exposure to either accidental or "acceptable" levels of petroleum hydrocarbons in the occupational environment.

Biotransformation enzymes in fish as tools for biomonitoring aquatic environment.

Biotransformation in fish--as in mammals--is catalyzed by several enzymes. These convert liposoluble endogenous and exogenous substrates to more water-soluble compounds prior to excretion. The biotransformation enzymes are induced by environmental pollutants. The induction can be expected to precede the onset of more serious changes at higher organization levels. We have studied the effect of petroleum from a ship spill and bleached kraft mill effluent on hepatic biotransformation enzyme activities of local fish species perch (Perca fluviatilis) and vedace (Coregonus albula) in Finland. Four months after the petroleum spill an elevated level of monoxygenase as well as glutathione S-transferase enzyme activities was seen in perch. Afterwards the difference between the control perch and the exposed ones disappeared. Bleached kraft mill effluent had effect on hepatic biotransformation in vendace. Increasing exposure time and effluent concentration elevated the activities.

[Comparative study of the behavior of particulate emissions from diesel and gasoline engines in animal lungs: elimination rate and induction of benzo(a)pyrene hydroxylase and ethoxycoumarin de-ethylase]. / Vergleichende Untersuchung zum Verhalten von partikulären Diesel- und Ottomotoremissionen in Tierlungen: Eliminationsrate und Induktion der Benzo(a)pyren Hydroxylase und Ethoxycumarin Deethylase.

The emitted particulates of five diesel-engined and two gasoline-engined passenger cars were investigated for the elimination rate from hamster lungs after intratracheal instillation. In addition extracts of these particulates were studied for their influence on the mixed function oxidase activity (MFO; Benzo(a)pyrene Hydroxylase, Ethoxycoumarine Deethylase). Differences in the elimination rates of diesel soot and particulates from gasoline engines were not found. Compared with the blanc the extracts of diesel soot from two vehicles proved to give a moderate increase of the MFO activity, but a significant difference to the blanc was observed with the extracts of the gasoline engines. It should be mentioned that the effects were studied without taking into account the quantitative relations of the emissions in the ambient air. However, the amounts of particulates were extremely high in relation to the natural conditions. In the limits of our test model there is no indication of a higher toxicity of diesel-emissions.

Low-dose chronic inhalation of diesel exhaust and/or coal dust by rats: effect of age and exposure on lung and liver cytochrome P-450.

Rats were exposed by inhalation to low levels of diesel exhaust and/or coal dust, seven hours/day, five days/week for 24 months. Cytochrome P-450-associated benzo[a]pyrene hydroxylase and 7-ethoxycoumarin deethylase activities were assayed in lung and liver microsomes after 3, 6, and 24 months. When data were analysed across all time intervals and adjusted for age, lower benzo[a]pyrene hydroxylase activity was observed in lung microsomes from rats exposed to diesel exhaust plus coal dust than in those exposed to coal dust alone. Data are discussed in terms of interaction between diesel exhaust and coal dust and effect of infectious agents.

The effects of dietary corn oil on the metabolism and activation of benzo[a]pyrene by the benzo[a]pyrene metabolizing enzymes of the mouse.

Male ICR Swiss mice, weighing 16-20 g, were fed ad libitum either a fat-free diet or a diet containing 10% corn oil. After three weeks on these diets, the rates of benzo[a]pyrene (B[a]P) metabolite formation and metabolism to products which covalently bind with macromolecules were compared using hepatic nuclei and microsomal preparations. The maximum activity of B[a]P hydroxylase in microsomes from untreated animals was increased 50% by feeding the corn oil diet, however, B[a]P hydroxylase in microsomes from 3-methylcholanthrene (3-MC)-treated mice was unaffected by diet. In animals treated with phenobarbital, B[a]P metabolism and B[a]P-DNA adduct formation were greater in microsomes from corn oil fed mice compared to those fed the fat-free diet. At a B[a]P concentration of 96 microM, microsomes from corn oil fed untreated mice produced 26% more extractable metabolites and covalent binding to exogenous DNA was increased 46%. At lower substrate concentrations (0.94-15.0 microM B[a]P), B[a]P-DNA and B[a]P-protein binding were 300-400% greater when incubated with microsomes from corn oil fed mice than when incubated with microsomes from mice fed fat-free diet. The apparent Vmax's determined for the formation of each extractable metabolite were increased 1.5-3.0 times by the corn oil diet. Hepatic nuclear B[a]P hydroxylase and nuclear activation of B[a]P to products which covalently bind to DNA in both non-induced and 3-MC-pretreated animals fed the corn oil diet were greater than that observed in animals fed the fat-free diet. B[a]P hydroxylase activities in the lungs of these animals were unaltered by diet.

Induction of mixed function oxidases by petroleum in the American eel, Anguilla rostrata.

American eels, Anguilla rostrata, were exposed to crude oil by ingestion of a 10, 100, or 500 microliters/kg fish dose per day for five days. Depuration was followed for an additional twelve days. All oil doses caused an induction of hepatic microsomal enzymes, maximally by three days of exposure. Benzo(a)pyrene hydroxylase (BaPH) showed a dose related response, with greater induction at 100 microliters/kg than at the other doses. The highest dose was hepatotoxic. Cytochrome P-450 induction was dose independent, and remained induced maximally for the entire experimental period, in contrast to BaPH which declined in activity. Reaction optimum for BaPH was at pH 7.5 and 27 degrees C. A study of tissue distribution showed the liver to account for nearly all BaPH activity. A significant increase in the protein content of the hepatic postmitochondrial fraction of oil-exposed fish was also observed.

Enhanced pulmonary and intestinal activation of procarcinogens and mutagens after chronic ethanol consumption in the rat.

Recent epidemiological surveys have indicated that alcoholics exhibit increased incidences of a variety of cancers. We have investigated, as a possible contributing factor to carcinogenesis in this population, the effect of chronic ethanol consumption on metabolic activation of procarcinogens by microsomes isolated from lungs and small intestine. These tissues are major sites through which procarcinogens enter the body and are also potential sites of procarcinogen metabolism. Rat litter-mates were pair-fed nutritionally adequate liquid diets which contained either ethanol as 36% of total energy or an equivalent energy content of carbohydrates in place of ethanol. Chronic ethanol consumption produced significant increases in pulmonary microsomal cytochrome P-450 and microsomal ethanol oxidation. The ethanol diet also enhanced the capacity of pulmonary microsomes to activate compounds present in tobacco pyrolyzates to mutagens detectable in the Ames Salmonella auxotroph reversion assay. The ethanol diet did not alter the capacity of pulmonary microsomes to hydroxylate benzo(a)pyrene (BaP) or to activate BaP to a mutagen. In contrast, microsomes from the upper small intestine of ethanol-fed rats did exhibit both higher levels of BaP hydroxylase activity and enhanced activation of BaP to a mutagen. The ethanol feeding also enhanced the capacity of the intestinal microsomes to activate to mutagens both tryptophan pyrolyzate and 2-aminofluorene but did not influence the metabolic activation of these promutagens by pulmonary microsomes. Chronic ethanol consumption thus influences carcinogen metabolism in the intestine and lung in a manner which varies with respect to both carcinogen and tissue.

Effects of dietary polychlorinated biphenyls on vitamin E and selenium nutrition in the chick.

Experiments were conducted to determine the effects of polychlorinated biphenyls (PCBs) on vitamin E-selenium nutrition in the chick. Results showed that 10 p.p.m. Aroclor¿ 1254 in the diets of breeding S.C.W.L. hens increased the susceptibility of progency to vitamin E-selenium deficiency when those chicks were reared on a diet deficient in vitamin E and supplemented with a marginal level of selenium. Susceptibility to this deficiency, as measured by the incidence of exudative diathesis, was also increased when PCBs were added to chick diets. Dietary PCBs were shown to induce hepatic microsomal benzopyrene hydroxylase and induction of this activity was associated with decreased biological utilization of dietary selenium. PCBs were shown to increase the apparent requirements of the chick for vitamin E and selenium for prevention of exudative diathesis. However, discrimination between effects on vitamin E function and effects on selenium function was not possible in these experiments.