Eicosanoid synthesis in 7,12-dimethylbenz(a)anthracene-induced mammary carcinomas in Sprague-Dawley rats fed primrose oil, menhaden oil or corn oil diet.

The comparative effects of high-fat diets (20%, w/w) on eicosanoid synthesis during mammary tumor promotion in 7,12-dimethylbenz(a)anthracene (DMBA)-induced rats were studied using diets containing 20% primrose oil (PO), 20% menhaden oil (MO) or 20% corn oil (CO). Sprague-Dawley rats fed the PO or MO diet had 21% of 24% fewer adenocarcinomas, respectively, than rats fed the CO diet. Histologically (i.e., mitotic figures, inflammatory cell infiltration and necrosis), the CO-fed rats exhibited the highest frequency of changes within tumors. Plasma fatty acid composition was significantly altered by diet, reflecting the composition of the oils which were being fed. Only the plasma of PO-fed rats contained detectable levels of gamma-linolenic acid (GLA). Arachidonic acid (AA) levels were significantly higher (p less than 0.05) in PO-fed than in CO- or MO-fed rats. MO-fed rats had significantly higher levels of plasma palmitic acid, while palmitoleic, eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids were detected only in MO-fed rats. As expected, linoleic acid (LA) and AA levels were lower (p less than 0.05) in the MO-fed rats than in PO- or CO-fed groups. The plasma of the CO-fed rats contained significantly higher levels of oleic acid. Eicosanoid synthesis in mammary carcinomas of rats fed the 20%-fat diets was 2-10 times higher than in mammary fat pads of control rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of 7,12-dimethylbenz(a)anthracene administration on hepatic drug metabolizing enzymes in female rats fed corn, primrose, and menhaden oils.

Female rats receiving a diet containing 20% menhaden oil beginning at 10 weeks of age and continuing for 13 weeks had hepatic benzo(a)pyrene [B(a)P] hydroxylase activity significantly higher than similar rats fed diets containing 20% corn oil or 20% oil of evening primrose. Compared to microsomes recovered from rats fed the corn oil diet, a significant increase in microsomal cytochrome P-450 content along with an increase in the activity of cytochrome P-450 mediated ethoxycoumarin O-dealkylase was evident in rats fed menhaden oil. Glutathione S-transferase activity of the cytosol of hepatocytes was increased twofold by the feeding of 20% menhaden oil, compared with the feeding of corn or primrose oil. Administration of 7,12-dimethylbenz(a)anthracene (DMBA) 21 days before instituting the diets enhanced B(a)P hydroxylase in all animals, with the activity greatest in those fed the menhaden oil. DMBA also caused a significant increase in ethoxycoumarin O-dealkylase in rats fed menhaden oil.

Phenobarbital depression of hepatic microsomal benzo[a]pyrene hydroxylation in rats starved and refed a diet containing menhaden fish oil: substrate and fat level dependency.

The influence of phenobarbital on the activity of hepatic mixed function oxidases responsible for benzo[a]pyrene hydroxylation was studied in rats fed diets containing menhaden fish oil (rich in n-3 fatty acids). Male rats were starved for 2 days and refed diet devoid of fat or containing 0.5, 10, or 20% menhaden oil for 4 days. Phenobarbital increased the apparent Km value as well as Vmax for benzo[a]pyrene hydroxylase in microsomes from rats fed the 20% menhaden oil diet. The increased Km was due to a progressive decrease in benzo[a]pyrene metabolism at the lower substrate concentrations, even in the presence of increased cytochrome P-450 content. The phenobarbital-induced increase in Km and the decreases in benzo[a]pyrene hydroxylation were not observed in rats fed 0.5% menhaden oil or a diet devoid of fat.

BP metabolism and the in vivo binding to hepatic DNA of rats fed diets containing menhaden fish oil.

It is well established that dietary polyunsaturated omega-6 fatty acids promote tumorigenesis and support the activity of mixed-function oxidases (MFO), which are responsible for carcinogen activation. This study was undertaken to determine if increased levels of dietary menhaden fish oil (rich in omega-3 fatty acids) would affect in vivo binding of [3H]benzo[a]pyrene (BP) to rat liver DNA. The effects of dietary menhaden oil on the activities of phase I and phase II drug-metabolizing enzymes were also studied to determine the possible relationships between the activity of these enzymes and the binding of [3H]BP metabolites to liver DNA. Following a single intraperitoneal injection of [3H]BP, more BP was bound to liver DNA recovered from rats fed diet containing 20% menhaden oil for 11 days at all time intervals tested (16, 24, 48, and 192 hours) than was bound from rats fed 0.5% menhaden oil. The increased binding of [3H]BP to liver DNA of rats fed the high level of menhaden oil may be due, in part, to increases in the MFO responsible for BP activation (as suggested by increased cytochrome P-450 level and total BP hydroxylase activity). The higher concentrations of radioactivity extracted from blood of rats fed 20% menhaden oil diet at the initial time periods (16 and 24 hours) may be a reflection of the greater capacity of the liver to metabolize BP to more water-soluble metabolites. The maximum velocity (Vmax) for ethoxycoumarin O-dealkylase, expressed as nanomoles per milligram protein or per gram liver, was increased three- to fourfold by feeding the high level of menhaden oil. The differences in these enzymatic responses suggest that certain form(s) of cytochrome P-450 are preferentially increased by feeding the omega-3 fatty acids of menhaden oil.

Influence of dietary menhaden oil on the enzymes metabolizing drugs and carcinogens.

Dietary polyunsaturated fatty acids enhance activation of constitutive and induced forms of enzymes of endoplasmic reticulum responsible for drug and carcinogen metabolism. The current report demonstrates that diets containing 10% or 20% refined menhaden fish oil that contains high concentrations of omega-3 fatty acids also supports these enzymes in a manner similar to that of oils that contain high concentrations of the omega-6 fatty acid linoleate. Cytosolic glutathione S-transferase was unaffected by dietary menhaden oil. However, ingestion of increasing concentrations of menhaden oil increased hepatic microsomal cytochrome P-450 content and the apparent Vmax for ethylmorphine N-demethylase, N-nitrosodimethylamine (DMN) N-demethylase, and benzo(a)pyrene [B(a)P] hydroxylase. Feeding menhaden oil increased the Km for ethylmorphine N-demethylase, and decreased Km's for DMN N-demethylase and B(a)P hydroxylase. Phenobarbital induced glutathione S-transferase activity only in rats fed 10% or 20% menhaden oil. Ethylmorphine N-demethylase was induced by only 25% by phenobarbital in rats refed the fat-free diet compared to 128% in rats refed the 20% menhaden oil. In contrast, DMN N-demethylase was induced only in rats fed the fat-free diet. B(a)P hydroxylase was induced in all rats regardless of the level of dietary fat. The specific activity of cytochrome P-450 for the metabolism of DMN and B(a)P, however, was significantly reduced in menhaden oil-fed animals by phenobarbital. This coupled with the increased Km for these reactions may have significant effects on the in vivo activation of these carcinogens in animals fed menhaden oil and subjected to dietary inducers of the mixed function oxidases.

In vitro activation and resultant binding of benzo[a]pyrene to DNA by microsomes from rats fed corn and menhaden oils.

Dietary unsaturated fat is required for maximum induction of the hepatic mixed function oxidases (MFO) responsible for activating carcinogens to forms that may bind covalently to DNA. The aim of this study was to assess the influence of dietary fat type and content on the activities of some enzymes involved in activation and detoxification of the carcinogen benzo[a]pyrene (B[a]P). Modification of these changes by pretreatment with phenobarbital (PB) was also evaluated. Male rats were fed diet devoid of fat or containing 20% corn oil (CO) or 20% menhaden fish oil (MO) for 4 days. PB induced soluble glutathione S-transferase, a detoxifying enzyme, only in rats fed dietary fats. Microsomes from rats fed both types of dietary fat had increased levels of cytochrome P-450 (P-450) and PB induced P-450 only in rats fed these fats. Although ethoxycoumarin O-dealkylase was significantly elevated in the MO group, the induction by PB was not dependent on dietary fat type or level. Dietary fat increased microsome-catalyzed in vitro binding of [3H]-B[a]P to calf thymus DNA, especially in response to PB. Menhaden oil depressed B[a]P hydroxylation and PB treatment depressed this activity to the greatest extent in rats fed this diet. When calculated as B[a]P metabolized per unit of P-450, PB seems to induce a P-450 in fat fed animals having lower affinity and capacity for B[a]P hydroxylation and activation than in rats fed the fat-free diet.