Increments of dietary linoleate raise liver arachidonate,but markedly reduce heart n−6 and n−3 fatty acids in the rat

Four diets containing 20% of energy (en%) as fat and with linoleic acid contents of 1.9, 3.1, 7.7 and 10.1 en%, respectively, were fed to one-month-old male rats for three months. The fatty acid profiles and the levels of the major n−6 and n−3 fatty acids in the lipids of plasma, liver, heart and kidney were measured. We found that with increasing concentrations of 18∶2n−6 in the diet, linoleic acid rose in plasma and in all organs, but long-chain n−6 and n−3 fatty acids responded differently. In liver, arachidonic acid increased and n−3 fatty acids were not significantly affected; in heart, both arachidonic and docosahexaenoic acids were progressively reduced; and in kidney, there was no change of n−6 and n−3. The results indicate that incremental changes in dietary, linoleate affect the levels of polyunsaturated fatty acids in liver and extrahepatic organs differently.     

Low-fat, monounsaturate-rich diets reduce susceptibility of low density lipoproteins to peroxidation ex vivo

Oxidative modification of low density lipoprotein (LDL) plays an important role in the process of atherosclerosis. The susceptibility of LDL to oxidation and the amount of peroxidation products formed are influenced by the lipoprotein content of 18∶1 n−9, 18∶2n−6, and the 18∶2n−6/18∶1n−9 ratio, which is dependent in part on dietary fatty acids. The purpose of this study was to determine if changing from a typical American diet to a low-fat, monousaturate-rich diet (LFMR) would result in favorable alterations in the fatty acid composition and oxidative profile of LDL in hypercholesterolemic individuals. Free-living postmennopausal hypercholesterolemic women who routinely consumed a diet moderately high in total fat and total saturates (34 and 11%, respectively) followed an LFMR diet (26% fat, 6% saturated fat, and 14% monounsaturated fat) for 6 mon. Sixteen postmenopausal hypercholesterolemic women already following standard low-fat (LF) diets acted as a control for seasonal variations in serum lipids. LDL from randomly selected subjects (LF n=6, LFMR n=5) was evaluated. LFMR diets resulted in LDL with increased concentrations and percentages of 18∶1n−9, reduced 18∶2n−6/18∶1n−9 ratio, and lower percentages of 18∶2n−6. No significant changes in LDL fatty acids occurred in the LF group. Conjugated diene lag time increased in both groups during copper-induced in vitro oxidation. Only the LFMR group experienced an increase in lipid peroxide lag time and a decrease in lipid peroxide formation. The LFMR diet was well tolerated and may be of therapeutic value in the treatment of hypercholesterolemia. A portion of this material was presented earlier at the annual meeting of the American Oil Chemists' Society and in abstract form [O'Byrne, D.J., Shireman, R.B., Knauft, D. (1993) The effects of a low-fat/high oleic acid diet on lipoproteins in postmenopausal hypercholesterolemic women, INFORM 4:553,#SS7].     

The effect of short-term diets rich in fish, red meat, or white meat on thromboxane and prostacyclin synthesis in humans

Foods which increase tissue arachidonic acid levels have been proposed to increase thrombosis tendency, presumably through increased platelet aggregation. This study examined the effect of doubling the dietary arachidonic acid (20∶4n−6) using meat- or fish-based diets on the systemic production of prostacyclin (PGl₂) and thromboxane (TXA₂) in 29 healthy, nonsmoking adults. There were three, 3-wk low-fat dietary periods (<15% energy as fat) in which subjects consumed a vegetarian diet for 1 wk followed by 2 wk on diets containing meat or fish as sources of 20∶4n−6. Between each diet period, there was a 3-wk washout period, during which subjects returned to their normal diets. The level of 20∶4n−6 consumed during the last 2 wk of each study was approximately double the usual intake (mean 140 mg/d), while the mean eicosapentaenoic acid (20∶5n−3) content of the diets varied from 1 mg/d on the white meat diet to 70 mg/d on the red meat diet and to 847 mg/d on the fish diet. The serum phospholipid (PL) 20∶4n−6/20∶5n−3 ratios were 11∶1 on the vegetarian diet, 15∶1 on the white meat diet, 8∶1 on the red meat diet, and 2∶1 on the fish diet (P<0.001). Neither white nor red meat diets affected platelet 20∶4n−6 levels, platelet aggregation, ex vivo platelet TXB₂ production, or the systemic PGl₂ or TXA₂ production as measured by gas chromatography-mass spectrometry analysis of the excretion levels of the principal urinary metabolites 2,3-dinor-6-keto-PGF_1α (PGl₂-M) and 11-dehydro-TXB₂ (TXA₂-M), respectively. The fish diet decreased the 20∶4n−6/20∶5n−3 ratio in platelet PL from the baseline level of 45∶1 to 13∶1 (P<0.001), had no effects on platelet aggregation, but significantly decreased platelet TXB₂, production (collagen-stimulated) and TXA₂-M production, while PGl₂-M levels were unaltered. These results indicate that short-term diets which double the usual 20∶4n−6 intake using white meat (175–330 g/d) or red meat (275–530 g/d) are not associated with an increased TXA₂ production, but this does not rule out the adverse effects of 20∶4n−6 at higher levels in the diet, or for more prolonged periods. Short-term diets containing fish (100–200 g/d with 90–210 mg/d 20∶4n−6 and approximately 650–1000 mg/d 20∶5n−3) led to significant increases in platelet 20∶5n−3 levels and a decrease in the ex vivo and systemic TXA₂ production.     

Comparison of the effects of dietary fish oils with different n−3 polyunsaturated fatty acid compositions on plasma and liver lipids in rats

The effects of dietary fish oils with different n−3 polyunsaturated fatty acid compositions on plasma lipid profiles in rats have been studied. Forty-eight male rats, previously maintained on a cholesterol-free diet for 15 days, were fed for 60 days with diets supplemented with 10% fat of either marine hilsa fish (Hilsa ilisa, family clupeidae) or fresh-water chital fish (Notopterus chitala, family notopteridae). The diets had similar levels of total saturated (35–41%), monounsaturated (43–47%) and n−3 polyunsaturated (9–10%) fatty acids. Cholesterol contents of the diets were adjusted to 0.85%; γ-linolenic acid (3.3%) in chital oil and eicosapentaenoic acid (4.9%) in hilsa oil diets were the major n−3 contributors. The percentage of eicosapentaenoic acid in the chital oil diet was 0.57 times that of the hilsa oil diet, but the eicosapentaenoic (EPA) to arachidonic acid (AA) ratio in the latter (4.08) was 3.2 times that of the former (1.27). Sixty days of hilsa oil diet feeding decreased the levels of cholesterol (53.3±2.9 to 50.0±1.1 mg/dL), triacylglycerol (75.7±3.8 to 64.3±2.6 mg/dL) and phospholipid (55.8±1.5 to 51.7±3.1 mg/dL) in rat plasma. Similar treatment with chital oil diet elevated the plasma cholesterol level (53.3±2.9 to 62.3±7.6 mg/dL) while triacylglycerol and phospholipid contents remained unaltered. Both the dietary treatments decreased the levels of linoleic and arachidonic acids in liver but only under the hilsa oil diet did the eicosapentaenoic acid percentage increase markedly (0.8±0.06% to 5.5±0.06%) at the expense of arachidonic acid. This study strongly suggests that the hypolipidemic effect depends on the composition of the n−3 polyunsaturated fatty acids rather than on the total n−3 polyunsaturated fatty acid content of the dietary fish oil.     

Incorporation of n−3 fatty acids into plasma and liver lipids of rats: Importance of background dietary fat

The health benefits of long-chain n−3 PUFA (20∶5n−3 and 22∶6n−3) depend on the extent of incorporation of these FA into plasma and tissue lipids. This study aimed to investigate the effect of the background dietary fat (saturated, monounsaturated, or n−6 polyunsaturated) on the quantitative incorporation of dietary 18∶3n−3 and its elongated and desaturated products into the plasma and the liver lipids of rats. Female weanling Wistar rats (n=54) were randomly assigned to six diet groups (n=9). The fat added to the semipurified diets was tallow (SFA), tallow plus linseed oil (SFA-LNA), sunola oil (MUFA), sunola oil plus linseed oil (MUFA-LNA), sunflower oil (PUFA), or sunflower oil plus linseed oil (PUFA-LNA). At the completion of the 4-wk feeding period, quantitative FA analysis of the liver and plasma was undertaken by GC. The inclusion of linseed oil in the rat diets increased the level of 18∶3n−3, 20∶5n−3, and, to a smaller degree, 22∶6n−3 in plasma and liver lipids regardless of the background dietary fat. The extent of incorporation of 18∶3n−3, 20∶5n−3, and 22∶5n−3 followed the order SFA-LNA>MUFA-LNA>PUFA-LNA. Levels of 22∶6n−3 were increased to a similar extent regardless of the type of major fat in the rat diets. This indicates that the background diet affects the incorporation in liver and plasma FA pools of the n−3 PUFA with the exception of 22∶6n−3 and therefore the background diet has the potential to influence the already established health benefits of long-chain n−3 fatty acids.     

Dietary fish oil dose- and time-response effects on cardiac phospholipid fatty acid composition

Fish consumption is associated with reduced cardiovascular mortality, and elevated myocardial long-chain n−3 polyunsaturated FA (PUFA) content is implicated in this cardioprotection. This study examined the dose and time responses for incorporation of n−3 PUFA into cellular membranes in rats fed fish oil (FO)-containing diets. For the time course study, rats were fed a 10% FO diet for periods ranging from 0 to 42 d, after which myocardial and erythrocyte membrane fatty acid composition was determined. For the dose response study, rats (n=3) were fed 0, 1.25, 2.5, 5, or 10% FO for 4 wk, with myocardial, erythrocyte, and skeletal muscle membrane FA determined. Myocardial DHA (22∶6n−3) levels doubled in 2 d, stabilizing at levels ≈200% higher than control after 28 d feeding with 10% FO. By comparison, DHA levels doubled after 4 wk of 1.25% FO feeding. In myocardium and skeletal muscle, EPA (20∶5n−3) levels remained low, but in erythrocytes EPA levels reached 50% of DHA levels. The n−3 PUFA were incorporated at the expense of n−6 PUFA in myocardium and skeletal muscle, whereas erythrocytes maintained arachidonic acid levels, and total n−3 PUFA incorporation was lower. This study shows that low doses of FO produce marked changes in myocardial DHA levels; maximal incorporation takes up to 28 d to occur; and while erythrocytes are a good indicator of tissue n−3 incorporation in stable diets, they vary greatly in their time course and pattern of incorporation.     

The effect of a salmon diet on blood clotting,platelet aggregation and fatty acids in normal adult men

This study was designed to measure the effect of dietary n−3 fatty acids (FA) on platelets and blood lipids. Healthy men (n=9), ages 31 to 65, were fed diets in which salmon was the source of n−3 fatty acids. They were confined in a nutrition suite at this Center for 100 days. Food intake and exercise levels were rigidly controlled. Initially they were placed on a stabilization diet for 20 days, then six men were fed the salmon diet for 40 days. The others remained on the stabilization diet. The two groups switched diets for the last 40 days of the study. Both diets were isocaloric [16% protein, 54% carbohydrate, and 30% fat by energy-% (En%)]. The salmon diet contained 7.5% of calories from n−6 FA and 2% from n−3 FA, primarily eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in a 40∶60 ratio, while the stabilization diet contained 7.5% of calories from n−6 FA and less than 0.3% n−3 FA, mainly 18∶3n−3. The bleeding time was unaffected by the diets in this study. The prothrombin time was shortened (11.6 sec.vs. 12.6 sec., p<0.01) for the subjects consuming the salmon diet as compared to that measured after 20 days of the stabilization diet. Mean platelet volume increased significantly during the period in which the volunteers consumed the salmon diet compared to the baseline diet (p<0.01), while the mean platelet levels decreased. Platelet aggregation (PA) was measured in platelet rich plasma before, during, and after the salmon diet using collagen, ADP, arachidonic acid (AA), and thrombin agonists. The PA threshold for ADP was significantly increased for the subjects on the salmon diet (p<0.05). No change in the PA threshold was detected for collagen or thrombin. The PA threshold for AA was unchanged also, but the platelets in subjects consuming the salmon diet had a prolonged time to maximum aggregation (p<0.01) with this reagent compared to platelets from men on the stabilization diet. Plasma, red cell, and platelet total FA composition was determined by capillary GLC. While the men consumed the salmon diets, there were marked increases (3 to 10-fold) in the EPA and DHA levels in all blood components with concomitant decreases in linoleic acid and AA levels. Thus, a salmon diet, high in n−3 FA, did not influence the bleeding times, but it decreased the sensitivity of platelets to ADP and AA, increased the mean platelet size, decreased the platelet count, and changed the FA composition of the plasma, RBC and platelet membrane lipids.     

Fatty acid carbon isotope ratios in humans on controlled diets

Carbon stable isotope ratios for six serum fatty acids (FA) are reported for human subjects on controlled fat diets to determine the range of natural isotope abundance and to demonstrate the leveling effect of a well-controlled diet. Twenty-nine subjects were randomly assigned to one of three controlled diets containing high, medium, or low fat. Diets were consumed for 8 wk. Serum samples were collected at baseline (0), 5, 6, 7, and 8 wk. FA were extracted and methylated. Isotope ratios were analyzed by high-precision gas chromatography combustion-isotope ratio mass spectrometry. At baseline, mean δ¹³C for 16:0^b, 16:1^a, 18:0^a, 18:1^c, 18:2n−6^d, and 20:4n−6^bc were −24.1, −21.7, −21.6, −25.6, −29.6, and −25.0%., respectively, with an average standard deviation of 1.9%.. Most δ¹³C decreased during the diet period and appeared to have stabilized by week 5 at −25.3, −21.9, −22.3, −26.5, −30.1, and −24.5%., respectively. Between-subject variability decreased from 1.74 to 1.20%. on the controlled diets. Measurement variability was 0.53%.. The within-subject variability during weeks 5–8 was 0.57%. (range of 0.32–0.84%.), showing a minimum biological fluctuation on controlled diets. There was no diet group effect on δ¹³C of serum FA. Except for 18:2, the δ¹³C of experimental diets was lower than that of serum FA, consistent with observations in animals. These data show that carbon isotope ratios stabilize in response to controlled diets within 5 wk, reflecting the isotope ratio of their dietary source, and establish isotope ratio fluctuations for endogenous compounds for future studies.     

Variations of <Emphasis Type="Italic">trans</Emphasis> octadecenoic acid in milk fat induced by feeding different starch-based diets to cows

The impact of starch sources differing in their velocities of ruminal degradation on the milk fat of dairy cows was studied. The animals received diets containing a slowly degradable (potatoes) or rapidly degradable (wheat) starch concentrate (40% of the dry matter) in a total mixed diet. Milk fat was the only animal performance factor affected: Cows produced significantly less milk fat when fed the wheat diet than the potato diet (−3.3 g/kg, −122 g/d; P<0.05). With the wheat diet, milk fat was poorer in short-chain FA and richer in unsaturated longchain FA, especially in trans octadecenoic acid (4.4 vs. 2.7% of the total FA, P<0.05). A very large increase in the isomer trans-10 18∶1 (+1.46% of the total FA) was observed. Because no difference in volatile FA concentrations in the rumen was revealed, the increase in trans octadecenoic acids, and particularly the isomer trans-10 18∶1, was associated with the larger postprandial drop in ruminal pH with wheat. Similar concentrate levels and FA profiles in both diets indicated that the decrease in milk fat was due to changes in the ruminal environment. Quicker degradation of wheat starch, and hence a greater drop in pH with this diet associated with the absence of any effect on volatile FA, strengthen the hypothesis developed in the literature of enzyme inhibition via increased levels of trans octadecenoic acids, especially the trans-10 isomer. Hence, milk fat can be decreased with rapidly degradable starch sources and not only with high levels of concentrates in the diet or added fat. More detailed work is necessary to elucidate the microorganisms involved and to determine whether metabolic pathways similar to those reported for high-concentrate diets are involved.     

Fatty acid composition of habitual omnivore and vegetarian diets

High-fat diets are implicated in the onset of cardiovascular disease (CVD), cancer, and obesity. Large intakes of saturated and trans FA, together with low levels of PUFA, particularly long-chain (LC) omega-3 (n−3) PUFA, appear to have the greatest impact on the development of CVD. A high n−6∶n−3 PUFA ratio is also considered a marker of elevated risk of CVD, though little accurate data on dietary intake is available. A new Australian food composition database that reports FA in foods to two decimal places was used to assess intakes of FA in four habitual dietary groups. Analysis using the database found correlations between the dietary intakes of LC n−3 PUFA and the plasma phospholipid LC n−3 PUFA concentrations of omnivore and vegetarian subjects. High meat-eaters (HME), who consumed large amounts of food generally, had significantly higher LC n−3 PUFA intakes (0.29 g/d) than moderate meat-eaters (MME) (0.14 g/d), whose intakes in turn were significantly higher than those of ovolacto-vegetarians or vegans (both 0.01 g/d). The saturated FA intake of MME subjects (typical of adult male Australians) was not different from ovolacto-vegetarian intakes, whereas n−6∶n−3 intake ratios in vegetarians were significantly higher than in omnivores. Thus, accurate dietary and plasma FA analyses suggest that regular moderate consumption of meat and fish maintains a plasma FA profile possibly more conducive to good health.     

Dietary n−3 FA modulate long and very long chain FA content, rhodopsin content, and rhodopsin phosphorylation in rat rod outer segment after light exposure

A previous study has shown that the long and very long chain FA (VLCFA) content of the rat retina responds to changes in dietary n−6/n−3 ratio of the fat fed (1). The present study tested whether similar changes in these FA are associated with alterations in rhodopsin content and rhodopsin phosphorylation after light treatment. Weanling rats were fed diets containing 20% (w/w, 40% energy) fat with either high (4.8%, w/w) or low (1.2%, w/w) n−3 FA. After 6 wk of feeding, half of the animals in each group were exposed to light for 48 h at 350 lx or were kept in complete darkness. In the rod outer segment, the high n−3 diet treatment increased the level of 20∶5n−3 and 22∶6n−3 and reduced the levels of 20∶4n−6 and 24∶4n−6 in PC, PE, and PS. After the feeding of a high n−3 FA diet, total n−3 pentaenoic VLCFA from C₂₄ to C₃₄ increased in PC, whereas the n−6 tetra- and pentaenoic VLCFA decreased. No changes occurred in n−3 hexaenoic VLCFA regardless of the level of 22∶6n−3 in the diet. After light exposure, animals fed a high n−3 FA diet showed reduction in 22∶6n−3 as well as in n−6 and n−3 VLCFA in PC. FFA and TG fractions contained increased levels of both 20∶4n−6 and 22∶6n−3 after light exposure. Dark-adapted rhodopsin content and rhodopsin phosphorylation in the rod outer segment of rats fed the low n−3 FA diet were higher than in animals fed a high n−3 FA diet. After light exposure, animals fed the low n−3 FA diet lost more rhodopsin compared to animals fed the high n−3 FA diet, resulting in less phosphorylation of rhodopsin. Results indicate that the FA composition, rhodopsin content, and phosphorylation in visual cells is influenced by the dietary n−3 FA fed as well as by light exposure. The results also imply that 22∶6n−3 may not be the precursor for synthesis of hexaenoic VLCFA.     

Effect of fish oil on the fatty acid composition of human milk and maternal and infant erythrocytes

To examine the effect of fish oil supplementation on the fatty acid (FA) composition of human milk and maternal and infant erythrocytes, five lactating women were supplemented with 6 g of fish oil daily for 21d. Usual maternal diets contained 1,147 mg of total n−3 FA, with 120 mg from very long-chain (>C₁₈) n−3 FA. Supplementation increased dietary levels to 3,092 mg of total n−3 FA and 2,006 mg of very long-chain n−3 FA. Milk samples were collected daily, prior to fish oil ingestion, and at 4-h intervals on days 1, 7, 14 and 21. Milk n−3 FA content increased within 8 h and reached steady state levels within one week. The n−6 fatty acid content decreased. Erythrocyte eicosapentaenoic acid content increased from 0.24% to 1.4% (P<0.01) in mothers and from 0.11% to 0.70% (P<0.05) in infants. Docosapentaenoic acid increased from 1.4% to 2.2% (P<0.05) in mothers and from 0.30% to 0.78% (P<0.01) in infants. There was no significant change in docosahexaenoic acid or n−6 fatty acid content. Maternal platelet aggregation responses were variable. No differences in milk or plasma tocopherol levels were noted. Based on a paper presented at the Symposium on Milk Lipids held at the AOCS Annual Meeting, Baltimore, MD, April 1990.     

Modulation of renal injury in pcy mice by dietary fat containing n−3 fatty acids depends on the level and type of fat

Low-fat diets and diets containing n−3 fatty acids (FA) slow the progression of renal injury in the male Han:Sprague-Dawley (SPRD)-cy rat model of polycystic kidney disease. To determine whether these dietary fat effects are similar in females and in another model of renal cystic disease, in this study we used both male and female pcy mice to examine the effects of fat level and type on disease progression. Adult pcy mice were fed 4, 10, or 20 g soybean oil/100 g diet for 130 d in study 1. In study 2, weanling pcy mice were fed high or low levels of fat rich in 18∶2n−6 (corn oil, CO) 18∶3n−3 (flaxseed oil/CO 4∶1 g/g, FO), or 22∶6n−3 (algal oil/CO 4∶1 g/g, DO) for 8 wk. In adult pcy mice, low-compared with high-fat diets lowered kidney weights (2.4±0.2 vs. 3.1±0.2 g/100 g body weight, P=0.006) and serum urea nitrogen (SUN) (9.6±0.6 vs. 11.9±0.6 mmol/L, P=0.009), whereas in young pcy mice it reduced renal fibrosis volumes (0.44±0.04 vs. 0.62±0.04 mL/kg body weight, P<0.0001). FO feeding in young pcy mice mitigated the detrimental effects of high fat on fibrosis while not altering kidney size, function, and oxidative damage when compared with the CO-fed mice. In contrast, DO-compared with CO-fed mice had higher kidney weights (2.64±0.07 vs. 2.24±0.08 g/100 g body weight, P=0.005), SUN (9.4±0.57 vs. 7.0±0.62 nmol/L, P<0.0001), and cyst volumes (7.9±0.28 vs. 6.2±0.30 mL/kg body weight, P<0.0001) and similar levels of oxidative damage and fibrosis. The FA compositions of the diets were reflected in the kidneys: 18∶2n−6, 18∶3n−3, and 22∶6n−3 were the highest in the CO, FO, and DO diets, respectively. Dietary effects on kidney disease progression were similar in males and females. A low-fat diet slows progression of renal injury in male and female pcy mice, consistent with findings in the male Han:SPRD-cy rat. Dietary fat type also influenced renal injury, with flaxseed oil diets rich in 18∶3n−3 slowing early fibrosis progression compared with diets rich in 18∶2n−6 or in 22∶6n−3.     

FA composition of plasma,red blood cell,and liver phospholipids of newborn piglets fed trans isomers of alpha-linolenic acid

The effects of dietary cis and trans α-linolenic acid (18∶3n−3) on the FA composition of plasma, red blood cell, and liver phospholipids were studied in newborn piglets. Animals were fed for 14 d with one of three diets: a control diet (group A) containing cis 18∶3n−3 at a level of 2.0% of total FA, a diet (group B) in which a part of the 18∶3n−3 acid was isomerized (1.3% of cis 18∶3n−3 and 0.7% of trans 18∶3n−3), or a diet (group C) with 2.0% cis 18∶3n−3 and 0.7% trans 18∶3n−3. Feeding animals with diets containing trans 18∶3n−3 resulted in the presence of trans isomers of 18∶3n−3, trans isomers of EPA, and trans isomers of DHA in phospholipids; however, the level of total trans n−3 PUFA in tissues was less than 0.3% of total tissue FA. In group B, the reduction of dietary amounts of cis 18∶3n−3 was associated with a decrease in individual and total cis n−3 PUFA. In contrast, in group C there was no decrease in tissue n−3 PUFA despite the increased dietary level of trans 18∶3n−3. These results suggest that the isomerization of a part of dietary n−3 PUFA, leading to the reduction of their levels in the diet, could induce a decrease in n−3 PUFA in phospholipids. The physiological effects of trans PUFA are not known and should be considered in future studies.     

Rat vitamin E status and heart lipid peroxidation: Effect of dietary α-Linolenic acid and marine n−3 fatty acids

Three groups of sixteen male rats each were fed semipurified diets containing 15% by weight of lipid for a period of 4 wk. The diets contained the same amount of polyunsaturated fatty acids (PUFA) (20% of total fatty acids) and saturated fatty acids (19% of total fatty acids). Dietary PUFA were represented exclusively by linoleic acid (18∶2 diet), or 10% linoleic acid and 10% linolenic acid (18∶3 diet), or 10% linoleic acid and 10% long-chain n−3 fatty acids (LCn−3 diet). The overall amount of vitamin E was similar in the three diets,i.e, 140, 133 and 129 mg/kg diet, respectively. Following appropriate extraction, tocopherol levels in heart, liver, brain, adipose tissue (AT) and plasma were measured by high-performance liquid chromatography. The level of vitamin E in the heart decreased with n−3 PUFA diets, most markedly with LCn−3 PUFA. Liver and AT vitamin E contents also decreased with n−3 PUFA diets when expressed as μg/mg total lipids and μg/mg phospholipids, respectively. Total plasma vitamin E was lower in rats fed the LCn−3 diet, but there was no significant difference when expressed as μg/mg total lipids. Brain vitamin E was not affected by the various diets.In vitro cardiac lipid peroxidation was quantified by the thiobarbituric acid reactive substances (TBARS) test. Heart homogenates were incubated at 37°C for 15 and 30 min in both the absence (uninduced) or presence (induced) of a free radical generating system (1 mM xanthine, 0.1 IU per mL xanthine oxidase, 0.2 mM/0.4 mM Fe/ethylenediaminetetraacetic acid). TBARS release was time-independent but significantly higher when LCn−3 fatty acids were fed to rats in either the uninduced or induced system. The study demonstrated that n−3 PUFA diets can influence vitamin E status of rats even in short-term experiments and can change the susceptibility of the heart toin vitro lipid peroxidation.     

Effects of a (n−3) polyunsaturated fatty acid-deficient diet on profiles of serum vitellogenin and lipoprotein in vitellogenic trout (Salmo gairdneri)

During the 6 months of vitellogenesis, 3-year-old female trout (Salmo gairdneri) were fed either an enriched (E) or an (n−3) polyunsaturated fatty acid (PUFA)-deficient (D) diet; serum vitellogenin (VG) and lipoproteins (d<1.21 g/ml) were analyzed at the third month of vitellogenesis (September) and at ovulation (December). The serum content of high density lipoproteins (HDL), the major protein class, maintained a mean value of 1500 mg/dl at both stages and with both diets. On the contrary, very low density lipoproteins (VLDL) were 90% higher during vitellogenesis than at spawning time, whereas excess vitellogenin circulated at this period (6580 mg/dl serum with diet E). The diet deficient in (n−3) lowered serum vitellogenin content by 16% in September and by 26% in December. The degree of (n−3) PUFA incorporation moderately decreased in low density lipoproteins (LDL) and in HDL with the (n−3)-deficient diet. The effect was more pronounced for 20∶5. On the other hand, essential 22∶6 was incorporated into vitellogenin at the same rate in September as in December with diet E (23% and 25%, respectively), whereas after a 3-month deficiency, the percentage fell to 12%; this percentage rose again to 19% at spawning time. These findings show that, although stored (n−3) PUFA were not exhausted after a 6-month dietary deficiency, the incorporation of essential fatty acids (EFA) into vitellogenin during the early stages of oogenesis was low, suggesting changes in egg composition that may influence hatching.     

Dietary modulation of fatty acid profiles and oxidative status of rat hepatocyte nodules: Effect of different n−6/n−3 fatty acid ratios

Male Fischer rats were fed the AIN76A diet containing varying n−6/n−3 FA ratios using sunflower oil (SFO), soybean oil (SOY), and SFO supplemented with EPA-50 and GLA-80 (GLA) as fat sources. Hepatocyte nodules, induced using diethylnitrosamine followed by 2-acetylaminofluorene/partial hepatoctomy promotion, were harvested, with surrounding and respective dietary control tissues, 3 mon after partial hepatectomy. The altered growth pattern of hepatocyte nodules in rats fed SFO is associated with a distinct lipid pattern entailing an increased concentration of PE, resulting in increased levels of 20∶4n−6. In addition, there is an accumulation of 18∶1n−9 and 18∶2n−6 and a decrease in the end products of the n−3 metabolic pathway in PC, suggesting a dysfunctional Δ-6-desaturase enzyme. The hepatocyte nodules of the SFO-fed rats exhibited a significantly reduced lipid peroxidation level that was associated with an increaser in the glutathione (GSH) concentration. The low n−6/n−3 FA ratio diets significantly decreased 20∶4n−6 in PC and PE phospholipid fractions with a concomitant increase in 20∶5n−3, 22∶5n−3, and 22∶6n−3. The resultant changes in the 20∶4/20∶5 FA ratio and the 20∶3n−6 FA level in the case of the GLA diet suggest a reduction of prostaglandin synthesis of the 2-series. The GLA diet also counteracted the increased level of 20∶4n−6 in PE by equalizing the nodule/surrounding ratio. The low n−6/n−3 ratio diets significantly increased lipid peroxidation levels in hepatocyte nodules, mimicking the level in the surrounding and control tissue while GSH was decreased. An increase in n−3 FA levels and oxidative status resulted in a reduction in the number of glutathione-S-transferase positive foci in the liver of the GLA-fed rats. Modulation of cancer development with low n−6/n−3 ratio diets containing specific dietary FA could be a promising tool in cancer intervention in the liver.     

The effect of dietary n−6 and n−3 polyunsaturated fatty acids on blood pressure and tissue fatty acid composition in spontaneously hypertensive rats

Effects of dietary n−6 and n−3 fatty acids (FAs) on blood pressure (BP) and tissue phospholipid (PL) FA composition in spontaneously hypertensive (SHR) and normotensive Wistar-Kyoto (WKY) rats were compared. Male weanling SHR and WKY were fed a fat-free semisynthetic diet supplemented with 10% (w/w) fats containing (a) 78% 18∶2n−6 (LA-rich), (b) 20% LA and 55% 18∶3n−3 (LN-rich), or (c) 11% LA and 3% LN (CON) for seven weeks. Dietary fats did not affect the BP elevation, but significantly altered the FA composition of brain, adrenal gland, renal medulla and cortex PL in SHR. The LA-rich diet increased n−6 FA while it reduced n−3 FA levels. The levels of 20∶4n−6 were not significantly different between animals fed the LA-rich and the CON diets. LN-rich diet increased the levels of n−3 FAs, while it reduced those of n−6 FAs. However, the extent of change was significantly less in SHR than in WKY. In all dietary groups, SHR, as compared to WKY, had a relatively higher level of the 2 series prostaglandin (PG) precursor, 20∶4n−6, and a relatively lower level of the 1 and 3 series PG precursors, 20∶3n−6 and 20∶5n−3. The possibility that the unbalanced eicosanoid FA precursor levels might contribute to the development of hypertension in this animal model is discussed.     

The Effects of dietary n−3/n−6 ratio on brain development in the mouse: a dose response study with long-chain n−3 fatty acids

This study examines the effects of the ratio of n−3/n−6 fatty acids (FA) on brain development in mice when longchain n−3 FA are supplied in the diet. From conception until 12 days after birth, B6D2F₁ mice were fed liquid diets, each providing 10% of energy from olive oil, and a further 10% from different combinations of free FA concentrates derived from safflower oil (18∶2n−6), and fish oil (20∶5n−3 and 22∶6n−3). The range of dietary n−3/n−6 ratios was 0,025, 0.5, 1.0, 2.0, and 4.0, with an n−6 content of greater than 1.5% of energy in all diets, and similar levels of total polyunsaturated fatty acids (PUFA). In an additional group of ratio 0.5, 18∶2n−6 was partially replaced by its δ6 desaturation product, 18∶3n−6. Biochemical analyses were conducted on 12-day-old pup brains, as well as on samples of maternal milk. No obvious effects on overall pup growth and development were observed, apart from a smaller litter size at ratio 1. Co-variance analysis indicated that increasing the n−3/n−6 ratio was associated with slightly smaller brains, relative to body weight. We found that 18∶2n−6 and 20∶5n−3 were the predominant n−6 and n−3 FA in the milk; in the brain these were 20∶4n−6 and 22∶6n−3, respectively. Increasing dietary n−3/n−6 ratios generally resulted in an increase in n−3 FA, with a corresponding decrease in n−6 FA. The n−3/n−6 ratio of the milk lipids showed a strong linear relationship with the diet, but in the brain the rate of increase tended to decrease beyond 0.5 (phosphatidylcholine, PC) and 0.25 (phosphatidylethanolamine, PE), such that there was a significant quadratic contribution to the relationship. The partial replacement of dietary 18∶2n−6 with 18∶3n−6 raised levels of 20∶4n−6 in milk, brain PC, and brain PE. These results indicate that the n−3/n−6 ratio of the phospholipids in the developing mouse brain responds maximally to maternal dietary long-chain n−3/n−6 ratios of between 0.25 and 0.5.     

Comparison of body weight and adipose tissue in male C57BI/6J mice fed diets with and withouttrans fatty acids

The effect of a diet containingtrans-fatty acids (tFA) on the fatty acid composition and fat accumulation in adipose tissue was investigated in mice. Male C57BI/6J mice were fed Control or Trans Diets that were similar, except that 50% of the 18∶1, which was allcis in the Control Diet, was replaced bytFA in the Trans Diet. At selected ages, body weight, epididymal fat pad weight, perirenal fat yield, adipose tissue cellularity and fatty acid composition were examined. Over the time period studied (2–24 mon), the proportion of 18∶0 and 16∶0 tended to decrease whilecis-18∶1 levels increased. Compared to the Control Diet, the Trans Diet resulted in adipose tissue lipids with higher percentages of 14∶0 and 18∶2n−6 and lower percentages ofcis-18∶1 and 20∶4n−6. In polar lipids,tFA replaced saturated fatty acids, whereastFA replacedcis-18∶1 in the nonpolar lipids. Body weights at 16 and 24 mon of age and epididymal fat pad weights at 8–24 mon of age were lower in mice fed the Trans Diet as compared to those fed the Control Diet. At the ages studied, the Trans Diet also resulted in lower values for perirenal fat weights, triacylglycerol to polar lipid ratios, and adipose cell size. The data suggest that chronic consumption oftFA affects lipid metabolism and results in decreased fat accumulation in murine adipose tissue.