Incorporation of n-3 fatty acids into plasma lipid fractions, and erythrocyte membranes and platelets during dietary supplementation with fish, fish oil, and docosahexaenoic acid-rich oil among healthy young men

The effects of n-3 fatty acid supplementation in the form of fresh fish, fish oil, and docosahexaenoic acid (DHA) oil on the fatty acid composition of plasma lipid fractions, and platelets and erythrocyte membranes of young healthy male students were examined. Altogether 59 subjects (aged 19–32 yr, body mass index 16.8–31.3 kg/m²) were randomized into the following diet groups: (i) control group; (ii) fish diet group eating fish meals five times per week [0.38±0.04 g eicosapentaenoic acid (EPA) and 0.67±0.09 g DHA per day]; (iii) DHA oil group taking algae-derived DHA oil capsules (1.68 g/d DHA oil group taking algae-derived DHA oil capsules (1.68 g/d DHA in triglyceride form); and (iv) fish oil group (1.33 g EPA and 0.95 g DHA/d as free fatty acids) for 14 wk. The fatty acid composition of plasma lipids, platelets, and erythrocyte membranes was analyzed by gas chromatography. The subjects kept 4-d food records four times during the study to estimate the intake of nutrients. In the fish diet, in DHA oil, and in fish oil groups, the amounts of n-3 fatty acids increased and those of n-6 fatty acids decreased significantly in plasma lipid fractions and in platelets and erythrocyte membranes. A positive relationship was shown between the total n-3 polyunsaturated fatty acids (PUFA) and EPA and DHA intake and the increase in total n-3 PUFA and EPA and DHA in all lipid fractions analyzed. DHA was preferentially incorporated into phospholipid (PL) and triglyceride (TG) and there was very little uptake in cholesterol ester (CE), while EPA was preferentially incorporated into PL and CE. The proportion of EPA in plasma lipids and platelets and erythrocyte membranes increased also by DHA supplementation, and the proportion of linoleic acid increased in platelets and erythrocyte membranes in the DHA oil group as well. These results suggest retroconversion of DHA to EPA and that DHA also interferes with linoleic acid metabolism.     

Cell Proliferation and Long Chain Polyunsaturated Fatty Acid Metabolism in a Cell Line From Southern Bluefin Tuna (Thunnus maccoyii)

Southern bluefin tuna (SBT, Thunnus maccoyii) aquaculture is a highly valuable industry, but research on these fish is hampered by strict catch quotas and the limited success of captive breeding. To address these limitations, we have developed a SBT cell line (SBT-E1) and here we report on fatty acid metabolism in this cell line. The SBT-E1 cells proliferated well in standard Leibovitz’s L-15 cell culture medium containing fetal bovine serum (FBS) as the source of fatty acids. Decreasing the FBS concentration decreased the cell proliferation. Addition of the C₁₈ polyunsaturated fatty acids (PUFA) α-linolenic acid (ALA, 18:3n-3) or linoleic acid (LNA, 18:2n-6) to the cell culture medium had little effect on the proliferation of the cells, whereas addition of the long-chain PUFA (LC-PUFA) arachidonic acid (ARA, 20:4n-6), eicosapentaenoic acid (EPA, 20:5n-3) or docosahexaenoic acid (DHA, 22:6n-3) significantly reduced the proliferation of the cells, especially at higher concentrations and especially for DHA. Addition of vitamin E to the culture medium overcame this effect, suggesting that it was due to oxidative stress. The fatty acid profiles of the total lipid from the cells reflected those of the respective culture media with little evidence for desaturation or elongation of any of the fatty acids. The only exceptions were EPA and ARA, which showed substantial elongation to 22:5n-3 and 22:4n-6, respectively, and DHA, which was significantly enriched in the cells compared with the culture medium. The results are discussed in light of the dietary PUFA requirements of SBT in the wild and in aquaculture.     

Comparative study between two different sources of n-3 poliunsaturated fatty acids and it effect on thymus and lipid profile in rats

In the present paper we analyzed the effect caused by different recovery diets enriched with n-3 polyunsaturated fatty acids (PUFA n-3) on thymus and serum lipid pattern. Severe depleted weanling Wistar rats (D) were divided in three groups that received during 10 days a 20% casein diet supplemented with EPA+DHA (group Cas), a 20% protein milk diet prepared using a commercial reduced-fat product enriched with linolenic and linoleic acids (group L) and a 20% casein diet as control group C. Cas and L gave each other 24 mg/day of PUFA n-3 being the ratio n-6/n-3 8.1/1 and 7.6/1, respectively. Thymus was removed and weighted and cell number were determined; blood was recollected and Total cholesterol, triacylglycerol, HDL and LDL-cholesterol fractions and myristic, palmitic, stearic, oleic, linoleic, linolenic, araquidonic, EPA and DHA fatty acid concentrations were measured in serum. Statistical analysis was performed using Anova test. Cell number were higher (p<0.01) in Cas (44.48+/-8.20) and in L (56.45+/-14.72) when compared to group D (1.80+/-0.70) and group C (23.70+/-4.04). L presented lower values of cholesterol, HDL and LDL-cholesterol (p<0.01) and higher values of triacylglycerol (p<0.05) when compared to Cas, being EPA (p<0.05) and DHA (p<0.01) higher in Cas. Being PUFA n-3 contribution the same in Cas and L, both diets were able to reverse the thymic athropy presenting a different hipolipemic behavior due to the different sources of PUFA n-3 used in the diets.     

Lipid Content and Fatty Acid Composition of 15 Marine Fish Species from the Southeast Coast of Brazil

Lipid content and fatty acid composition were determined in edible meat of fifteen marine fish species caught on the Southeast Brazilian coast and two from East Antarctic. Most of the fish had lipid amounts lower than 10% of their total weight. Palmitic acid (C16:0) predominated, accounting for 54–63% of the total amount of saturated fatty acids. Oleic acid (C18:1n-9) was the most abundant (49–69%) monounsaturated fatty acid, and docosahexaenoic acid (DHA) was the predominant polyunsaturated fatty acid (PUFA), accounting for 31–84% of n-3 PUFA. n-3 PUFA level were highest in Antarctic fish meat, comprising 45% of the total fatty acid content, which consisted of mainly EPA (16.1 ± 1.5 g/100 g lipids) and DHA (24.8 ± 2.4 g/100 g lipids). The amounts of EPA + DHA in g/100 g of lipids on the Southeast Brazilian coast and Antarctic fish species investigated were found to be similar: 42.0 ± 1.7 for Bonito cachorro, 41.0 ± 2.3 for Atum, and 39.4 ± 1.8 for peixe porco, respectively. All the studied species exhibited an n-3/n-6 ratio higher than 3, which confirms the great importance of Southeast Brazilian coast fish as a significant dietary source of n-3 PUFA.     

Differences in Lipid Characteristics Among Populations: Low-Temperature Adaptability of Ayu, Plecoglossus altivelis

The lipid and fatty acid compositions of the total lipids of three cultured populations (migratory between fresh and salt water, Lake Biwa landlocked, and Setogawa River forms) of ayu, Plecoglossus altivelis, were investigated to clarify the difference in lipid characteristics and temperature adaptability among the three groups. Triacylglycerols were the dominant depot lipids of the three populations, while phospholipids, such as phosphatidylcholine and phosphatidylethanolamine, were found to be the major components of the polar lipids, and their lipid classes are similar to each other. The major fatty acids in the triacylglycerols of all specimens were 16:0, 18:0, 16:1n-7, 18:1n-7, 18:1n-9, 18:2n-6 (linoleic acid), 20:5n-3 (EPA, icosapentaenoic acid), and 22:6n-3 (DHA, docosahexaenoic acid), similar to the tissue phospholipids of the three populations, 16:0, 18:0, 16:1n-7, 18:1n-7, 18:1n-9, 18:2n-6, 20:4n-6, EPA, and DHA. All classes had high levels of 18:2n-6, which originates from their dietary lipids. Compared with the lower DHA levels of the triacylglycerols, the higher levels in the phospholipids suggest their selective accumulation or a biosynthetic pathway to DHA as in freshwater fish. Two populations (the migratory and Setogawa River forms) adapted to lower temperatures with comparatively high levels of polyunsaturated fatty acids (PUFA) for their membrane fluidities. With significantly higher levels of n-3 PUFA and total PUFA, the mean DHA content in the lipids of the Setogawa River form (the population that adapted to lower temperatures) was significantly higher than that of the migratory form. From these results, we concluded that the Setogawa River population actively concentrates long-chain PUFA in its polar lipids and has high adaptability to low temperature.     

Interlaboratory Assessment of Dried Blood Spot Fatty Acid Compositions

Dried blood spots for fatty acid profiling are increasing in popularity; however, variability in results between laboratories has not been characterized. Whole blood from two subjects (low and high n-3 polyunsaturated fatty acid [PUFA] status) was collected, 25 μL applied to butylated hydroxytoluene (BHT)-treated chromatography strips, dried in air, and shipped to five laboratories. Results were reported as “routine” (typical fatty acids for each laboratory) or “standardized” (a set of 19 fatty acids), and outliers and variability (%CV) were determined. Five and eight outliers of a possible 91 measures each were identified by routine and standardized reporting, respectively, including eicosapentaenoic acid (EPA, 20:5n-3) in the low n-3 PUFA sample and arachidonic acid in the high n-3 PUFA sample. By standardized reporting, no outliers were identified for EPA or docosahexaenoic acid (DHA, 22:6n-3), and %CV decreased from 8.6% to 6.0% and 9.1% to 6.6% for EPA and 10.5% to 7.2% and 10.5% to 6.6% for DHA in the low and high n-3 PUFA sample, respectively. In conclusion, fatty acid profiles yielded few outliers, and standardization of reporting reduced the variability between laboratories.     

Oxidative stability of polyunsaturated fatty acids and soybean oil in an aqueous solution with emulsifiers

The oxidative stability of polyunsaturated fatty acids (PUFA) and soybean oil homogenized with emulsifiers was investigated. Model emulsions were prepared from PUFA, including linoleic acid (LA), arachidonic acid (AA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA), and from soybean oil emulsified with different emulsifiers: three Tween emulsifiers (Tween 20, Tween 60, Tween 80) and two sucrose esters (S-1170 and S-1570) were used. The results showed that the emulsions prepared from LA and the various emulsifiers, oxidized at 40°C, were unstable. However, the corresponding AA, EPA, and DHA emulsions were stable, indicating that PUFA with a higher degree of unsaturation were more stable with emulsifiers than without the emulsifiers. In the soybean oil-in-water model system, the oxidation of soybean oil with various emulsifiers was less than without the emulsifiers.     

Low-dose eicosapentaenoic or docosahexaenoic acid administration modifies fatty acid composition and does not affect susceptibility to oxidative stress in rat erythrocytes and tissues

In view of the promising future for use of n-3 polyunsaturated fatty acids (PUFA) in the prevention of cancer and cardiovascular diseases, it is necessary to ensure that their consumption does not result in detrimental oxidative effects. The aim of the present work was to test a hypothesis that low doses of eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA) do not induce harmful modifications of oxidative cell metabolism, as modifications of membrane fatty acid composition occur. Wistar rats received by gavage oleic acid, EPA, or DHA (360 mg/kg body weight/day) for a period of 1 or 4 wk. Fatty acid composition and α-tocopherol content were determined for plasma, red blood cell (RBC) membranes, and liver, kidney, lung, and heart microsomal membranes. Susceptibility to oxidative stress induced by tert-butylhydroperoxide was measured in RBC. EPA treatment increased EPA and docosapentaenoic acid (DPA) content in plasma and in all the membranes studied. DHA treatment mainly increased DHA content. Both treatments decreased arachidonic acid content and n-6/n-3 PUFA ratio in the membranes, without modifying the Unsaturation Index. No changes in tissue α-tocopherol content and in RBC susceptibility to oxidative stress were induced by either EPA or DHA treatment. The data suggest that EPA and DHA treatments can substantially modify membrane fatty acids, with-out increasing susceptibility to oxidative stress, when administered at low doses. This opens the possibility for use of low doses of n-3 PUFA for chemoprevention without risk of detrimental secondary effects.     

Concentration and purification of stearidonic,eicosapentaenoic, and docosahexaenoic acids from cod liver oil and the marine microalgaIsochrysis galbana

n-3 Polyunsaturated fatty acids (n-3 PUFA) from the marine microalgaIsochrysis galbana were concentrated and purified by a two-step process—formation of urea inclusion compounds followed by preparative high-performance liquid chromatography. These methods had been developed previously with fatty acids from cod liver oil. By the urea inclusion compounds method, a mixture that contained 94% (w/w) stearidonic (SA), eicosapentaenoic (EPA), plus docosahexaenoic (DHA) acids (4:1 urea/fatty acid ratio and 4°C crystallization final temperature) was obtained from cod liver oil fatty acids. Further purification of SA, EPA, and DHA was achieved with reverse-phase C₁₈ columns. These isolations were scaled up to a semi-preparative column. A PUFA concentrate was isolated fromI. galbana with methanol/water (80:20, w/w) or ethanol/water (70:30, w/w). With methanol/water, a 96% EPA fraction with 100% yield was obtained, as well as a 94% pure DHA fraction with a 94% yield. With ethanol/water as the mobile phase, EPA and DHA fractions obtained were 92% pure with yields of 84 and 88%, respectively.     

Headspace gas chromatography of volatile lipid peroxidation products from human red blood cell membranes

An improved headspace capillary gas chromatographic (GC) method was developed to measure the oxidative susceptibility of human red blood cell (RBC) membranes. This method analyzed volatile peroxidation products of both n−6 (hexanal and pentane) and n−3 (propanal) polyunsaturated fatty acids. Oxidative susceptibility tests were standardized by incubating in a sealed 10-mL headspace bottle 0.25 or 1 mL of human RBC membrane in 40 mM phosphate buffer for 1 hr at 37°C with a mixture of Fe⁺⁺, ascorbic acid and H₂O₂. Sodium dodecyl sulfate increased significantly the amount of hexanal measured by headspace GC. By this standard headspace method, in one series of red blood cell membranes (RBCM) samples a four-fold variation in oxidative susceptibility was observed in RBCM from blood freshly drawn from six healthy subjects. In another series of RBCM samples a sixteen-fold variation in oxidative susceptibility was noted in frozen RBCM from blood freshly drawn from five healthy subjects. Correlation between hexanal formation and polyunsaturated fatty acids (PUFA) depletion provided good evidence that under these standard conditions hexanal is exclusively derived from the oxidation of arachidonic acid. Hydroperoxides of arachidonic acid are more readily formed and decomposed than those of linoleic acid in the presence of Fe⁺⁺, ascorbic acid and H₂O₂ to produce hexanal as the main product that can be readily analyzed by headspace GC. This method may provide a useful tool to study susceptibility toward lipid peroxidative damage in human RBC membranes.     

Cholesterol,Lipid Content,and Fatty Acid Composition of Different Tissues of Farmed Cobia (<Emphasis Type="Italic">Rachycentron canadum</Emphasis>) from China

Marine fishes are rich in n-3 polyunsaturated fatty acids (PUFA), especially eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), which are extremely important for human health. The objective of our work was to determine the content and composition of lipids and fatty acids in the different tissues of cobia from China and to evaluate their nutritional value. The results showed that cobia from China was rich in lipids; the neutral lipid content was above 82%; the content of cholesterol and phospholipid was low. Eighteen fatty acids were identified. Myristic (C14:0), palmitic (C16:0), and stearic acids (C18:0) were the main saturated acids; palmitoleic (C16:1n-7) and oleic acid (C18:1n-9) were the main monounsaturated fatty acids. EPA and DHA were the main PUFA; n-3 and n-6 PUFA were present as 12–18% and 2.6–3.2% of the total fatty acids, respectively. The n-6/n-3 ratio was in the range from 0.18 to 0.22, which was far lower than that (5:1) recommended by WHO/FAO. Therefore, cobia lipids from China have a high nutritional value.     

N-3 Polyunsaturated Fatty Acids Modulate In-Vitro T Cell Function in Type I Diabetic Patients

In this work, we assessed the in-vitro effects of eicosapentaenoic acid (EPA; C20:5n-3) and docosahexaenoic acid (DHA; C22:6n-3) (final concentration, 15 microM) on T cell blastogenesis, interleukin-2 and -4 (IL-2, IL-4) secretion, fatty acid composition and intracellular oxidative status in type I diabetic patients with or without complications. Con A stimulated lymphocyte proliferation, glucose uptake, intracellular reduced glutathione levels and catalase activity were lower in diabetics as compared to controls, regardless to the presence of complications. EPA and DHA diminished T-lymphocyte proliferation and IL-2 production but enhanced IL-4 secretion in both diabetic and control groups. No changes in the levels of reduced glutathione and in the activities of catalase and SOD were observed in control T cells cultured in the presence of EPA and DHA. However, in diabetic patients, addition of n-3 PUFA to culture induced an increase in T cell levels of reduced glutathione and hydroperoxide, and in activities of catalase and SOD. Low levels of arachidonic acid (C20:4n-6) were found in plasma membrane phospholipids of lymphocytes from diabetic patients compared to controls. Incubation of lymphocytes with EPA and DHA was associated with an incorporation of these fatty acids in membrane phospholipids. In conclusion, the beneficial effects of n-3 PUFA on T cell functions in type I diabetes could be attributed to their suppressive action and modulation of cytokine secretion, and to the improvement of intracellular oxidative status.     

Accretion of Dietary Docosahexaenoic Acid in Mouse Tissues Did Not Differ between Its Purified Phospholipid and Triacylglycerol Forms

Recent studies suggest that dietary krill oil leads to higher omega-3 polyunsaturated fatty acids (n-3 PUFA) tissue accretion compared to fish oil because the former is rich in n-3 PUFA esterified as phospholipids (PL), while n-3 PUFA in fish oil are primarily esterified as triacylglycerols (TAG). Tissue accretion of the same dietary concentrations of PL- and TAG-docosahexaenoic acid (22:6n-3) (DHA) has not been compared and was the focus of this study. Mice (n = 12/group) were fed either a control diet or one of six DHA (1%, 2%, or 4%) as PL-DHA or TAG-DHA diets for 4 weeks. Compared with the control, DHA concentration in liver, adipose tissue (AT), heart, and eye, but not brain, were significantly higher in mice consuming either PL- or TAG-DHA, but there was no difference in DHA concentration in all tissues between the PL- or TAG-DHA forms. Consumption of PL- and TAG-DHA at all concentrations significantly elevated eicosapentaenoic acid (20:5n-3) (EPA) in all tissues when compared with the control group, while docoshexapentaenoic acid (22:5n-6) (DPA) was significantly higher in all tissues except for the eye and heart. Both DHA forms lowered total omega-6 polyunsaturated fatty acids (n-6 PUFA) in all tissues and total monounsaturated fatty acids (MUFA) in the liver and AT; total saturated fatty acid (SFA) were lowered in the liver but elevated in the AT. An increase in the DHA dose, independent of DHA forms, significantly lowered n-6 PUFA and significantly elevated n-3 PUFA concentration in all tissues. Our results do not support the claim that the PL form of n-3 PUFA leads to higher n-3 PUFA tissue accretion than their TAG form.     

Lipase-assisted concentration of n-3 polyunsaturated fatty acids in acylglycerols from marine oils

Preparation of n-3 polyunsaturated fatty acid (PUFA) concentrates from seal blubber oil (SBO) and menhaden oil (MHO) in the form of acylglycerols was carried out by hydrolysis with a number of commercial microbial lipases. The lipases tested were Aspergillus niger, Candida cylindracea (CC), Chromobacterium viscosum, Geotrichum candidum, Mucor miehei, Pseudomonas sp., Rhizopus oryzae, and Rhizopus niveus. After lipase-assisted hydrolysis of oils, free fatty acids were removed, and fatty acid composition of the mixture containing mono-, di-, and triacylglycerols was determined. All lipases were effective in increasing the n-3 PUFA content of the remaining acylglycerols of both SBO and MHO. The highest concentration of n-3 PUFA was provided by CC lipase; 43.5% in SBO [9.75% eicosapentaenoic acid (EPA), 8.61% docosapentaenoic acid (DPA), and 24.0% docosahexaenoic acid (DHA)] and 44.1% in MHO (18.5% EPA, 3.62% DPA, and 17.3% DHA) after 40 h of hydrolysis. Thus, CC lipase appears to be most suitable for preparation of n-3 PUFA in the acylglycerol form from marine oils.     

Production of PUFA Concentrates from Poultry and Fish Processing Waste

In fish and poultry processing, viscera are generally considered as a waste product and often discarded. Chicken and hilsa fish (Hilsa ilisa) viscera were used for the production of polyunsaturated fatty acids (PUFA) linoleic (18:2n-6), eicosapentaenoic (EPA; 20:5n-3) and docosahexaenoic acid (DHA; 22:6n-3). Free fatty acids (FFA) were extracted by alkaline hydrolysis of chicken and fish viscera; yields were 5.2 and 5.9% (w/w) respectively. PUFA concentrates were obtained by a two step process—deduction of saturated fatty acids (FA) by low temperature crystallization in acetone followed by urea inclusion compound-based fractionation. Acetone treatment removed 90 and 96% of saturated FA in chicken and fish viscera respectively with FA to acetone ratio of 1:12 (w/v). Using an urea to FA ratio (w/w) of 4.0, chicken viscera produced a maximum of 84.1% of PUFA concentrates containing 82.1% of linoleic acid with a yield of 10% where as in the case of fish viscera the maximum PUFA concentrates were 91.3% containing 78.2% of EPA-DHA with the yield of 11%. Thus, the utilization of poultry and fish processing waste into the production of PUFA concentrates has been shown.     

Matrix Extension Validation of AOCS Ce 2c-11 for Omega-3 Polyunsaturated Fatty Acids in Conventional Foods and Dietary Supplements Containing Added Marine Oil

Marine oils are commonly added to conventional foods and dietary supplements to enhance their contents of omega-3 polyunsaturated fatty acids (PUFA), including eicosapentaenoic acid (EPA; 20:5n-3) and docosahexaenoic acid (DHA; 22:6n-3), which have been associated with numerous potential health benefits. This study compared American Oil Chemists’ Society (AOCS) Official Methods Ce 2b-11 and Ce 2c-11 for determining EPA and DHA in foods and dietary supplements and found that AOCS Ce 2c-11 produces significantly higher analyzed values, which could be attributed to a more comprehensive breakdown of the sample matrix and derivatization of fatty acids. Our subsequent food matrix extension validation of AOCS Ce 2c-11 demonstrated that the method produces true, accurate, sensitive, and precise determinations of EPA, DHA, and total omega-3 PUFA in foods and dietary supplements containing added marine oil, including those formulated with emulsified and microencapsulated oils. The method detection limits for EPA and DHA were 0.012 ± 0.002 and 0.011 ± 0.003 mg g⁻¹, respectively (means ± SD). The analyzed contents of EPA (1.26–386 mg serving⁻¹), DHA (1.37–563 mg serving⁻¹), and total omega-3 PUFA (2.69–1270 mg serving⁻¹) were reported for 27 conventional food and dietary supplement products. Eighteen products declared contents of DHA, EPA + DHA, or total omega-3 PUFA on product labels, and the analyzed contents of those fatty acids varied from 95 to 162% of label declarations for all but two of the products.     

Polyunsaturated fatty acids exert antiarrhythmic actions as free acids rather than in phospholipids

Previous studies have shown that exogenous free n-3 polyunsaturated fatty acids (PUFA) can prevent tachyarrhythmias caused by specific agents in isolated cardiac myocytes. However, the question as to whether incorporation of the n-3 PUFA into membrane phospholipids has the same immediate protective effects remained to be answered. To answer this question, we increased the content of n-3 PUFA in the phospholipids of cultured neonatal rat myocytes by growing them 2–3 d in a culture to which eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA) in 15 μM concentration was added. Analysis of the fatty acid composition of membrane phospholipids revealed a significantly higher level of EPA and DHA (from 0.2 to 7.6% and from 1.2 to 6.5%) in cells supplemented with EPA or DHA, respectively. The responses of the myocytes grown in normal media or in media enriched with the PUFA to arrhythmogenic agents were examined after free fatty acids were removed from the medium and the cells. The arrhythmogenic agents used were the β-adrenergic agonist isoproterenol or an elevated extracellular concentration of calcium. The results showed that there was no significant difference in the induction of tachyarrhythmias by isoproterenol or by elevated [Ca²⁺]_o in cells grown in media enriched with PUFA, as compared with cells grown in normal media in the absence of the free PUFA. Under the conditions of this study, only the unesterified PUFA were able to protect the cardiomyocytes against induced arrhythmias. There was no antiarrhythmic effect due to an increased fraction of EPA or DHA in membrane phospholipids.     

Eicosapentaenoic acid,but not docosahexaenoic acid,increases mitochondrial fatty acid oxidation and upregulates 2,4-dienoyl-CoA reductase gene expression in rats

The aim of the present study was to investigate whether eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA) was responsible for the triglyceride-lowering effect of fish oil. In rats fed a single dose of EPA as ethyl ester (EPA-EE), the plasma concentration of triglycerides was decreased at 8 h after acute administration. This was accompanied by an increased hepatic fatty acid oxidation and mitochondrial 2,4-dienoyl-CoA reductase activity. The steady-state level of 2,4-dienoyl-CoA reductase mRNA increased in parallel with the enzyme activity. An increased hepatic long-chain acyl-CoA content, but a reduced amount of hepatic malonyl-CoA, was obtained at 8 h after acute EPA-EE treatment. On EPA-EE supplementation, both EPA (20:5n-3) and docosapentaenoic acid (DPA, 22:5n-3) increased in the liver, whereas the hepatic DHA (22:6n-3) concentration was unchanged. On DHA-EE supplementation retroconversion to EPA occurred. No statistically significant differences were found, however, for mitochondrial enzyme activities, malonyl-CoA, long-chain acyl-CoA, plasma lipid levels, and the amount of cellular fatty acids between DHA-EE treated rats and their controls at any time point studied. In cultured rat hepatocytes, the oxidation of [1-¹⁴C]palmitic acid was reduced by DHA, whereas it was stimulated by EPA. In thein vivo studies, the activities of phosphatidate phosphohydrolase and acetyl-CoA carboxylase were unaffected after acute EPA-EE and DHA-EE administration, but the fatty acyl-CoA oxidase, the rate-limiting enzyme in peroxisomal fatty acid oxidation, was increased after feeding these n-3 fatty acids. The hypocholesterolemic properties of EPA-EE may be due to decreased 3-hydroxy-3-methylglutaryl-CoA reductase activity. Furthermore, replacement of the ordinary fatty acids, i.e., the monoenes (16:1n-7, 18:1n-7, and 18:1n-9) with EPA and some conversion to DPA concomitant with increased fatty acid oxidation is probably the mechanism leading to changed fatty acid composition. In contrast, DHA does not stimulate fatty acid oxidation and, consequently, no such displacement mechanism operates. In conclusion, we have obtained evidence that EPA, and not DHA, is the fatty acid primarily responsible for the triglyceride-lowering effect of fish oil in rats.     

The Brain Oxylipin Profile Is Resistant to Modulation by Dietary n-6 and n-3 Polyunsaturated Fatty Acids in Male and Female Rats

Oxylipins are bioactive lipids formed by the monooxygenation of polyunsaturated fatty acids (PUFA). Eicosanoids derived from arachidonic acid (ARA) are the most well-studied class of oxylipins that influence brain functions in normal health and in disease. However, comprehensive profiling of brain oxylipins from other PUFA with differing functions, and the examination of the effects of dietary PUFA and sex differences in oxylipins are warranted. Therefore, female and male Sprague–Dawley rats were provided standard rodent diets that provided additional levels of the individual n-3 PUFA α-linolenic acid (ALA), eicosapentaenoic acid (EPA) or docosahexaenoic acid (DHA), or the n-6 PUFA linoleic acid (LNA) alone or with ALA (LNA + ALA) compared to essential fatty acid-sufficient control diets. Oxylipins and PUFA were quantified in whole brains using HPLC-MS/MS and GC, respectively. Eighty-seven oxylipins were present at quantifiable levels: 51% and 17% of these were derived from ARA and DHA, respectively. At the mass level, ARA and DHA oxylipins comprised 81–90% and 6–12% of total oxylipins, while phospholipid ARA and DHA represented 25–35% and 49–62% of PUFA mass, respectively. Increasing dietary n-3 PUFA resulted in higher levels of oxylipins derived from their precursor PUFA; otherwise, the brain oxylipin profile was largely resistant to modulation by diet. Approximately 25% of oxylipins were higher in males, and this was largely unaffected by diet, further revealing a tight regulation of brain oxylipin levels. These fundamental data on brain oxylipin composition, diet effects, and sex differences will help guide future studies examining the functions of oxylipins in the brain.