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Analytical-scale high-performance liquid chromatography of omega-3 fatty acid esters derived from fish oils.

Fish oil triglycerides were transesterified to their corresponding methyl and ethyl esters. The esters were separated by reversed-phase high-performance liquid chromatography on a 10-cm column of 5-microns octadecylsilyl silica, using a ternary mobile phase of acetonitrile-tetrahydrofuran-water (9:5:11). The separation of principal saturated and unsaturated C14-C22 fatty acid esters was accomplished in 60 min. For the polyenoic acid esters, detection limits of ca. 26 micrograms were observed, and baseline resolution of the important critical pair of esters of the omega 3 fatty acids, eicosapentaenoic acid (C20:5 omega 3) and docosahexaenoic acid (C22:6 omega 3) was achieved. However, to obtain baseline resolution of the esters from minor fatty acid esters, a urea crystallization procedure was required prior to chromatographic analysis. The influence on the reversed-phase chromatographic retention of four different ester moieties on the fatty acids were also studied. The n-alkyl esters exhibited an increase in ln k' with increasing chain length and with increasing lipophilic character of the alkyl ester moieties; however, with the aromatic benzyl derivatives there was a decrease in ln k' compared with the alkyl esters.

Chromatography, High Pressure Liquid↗

Renal cortex remodeling in streptozotocin-induced diabetic spontaneously hypertensive rats treated with olive oil, palm oil and fish oil from Menhaden.

We studied the effects of edible oils intake on the renal cortical structure of streptozotocin-induced diabetic (Db) and non-diabetic spontaneously hypertensive rats (SHR). Male SHR divided into 5 groups were studied during 6 weeks: one non-diabetic SHR group and four diabetic SHR groups (three groups received by gavage olive, palm or fish oil). Kidneys were analyzed by light microscopy and stereology. Oils intake did not change the plasma glucose levels. The blood pressure (BP) was lower in SHR-Db than in SHR, but SHR-Db-fish oil showed the lowest BP. Creatinine clearance was different between diabetic SHR and non-diabetic SHR, but not between treated SHR-Db and untreated SHR-Db. The renal cortex showed scars surrounding obsolete glomeruli with inflammatory infiltrate mainly in untreated SHR-Db. The olive oil, palm oil and mainly fish oil intake retard the usual loss of glomeruli and attenuate the renal cortex adverse remodeling of Db and non-Db SHR.

Animals↗

Dietary effects of arachidonate-rich fungal oil and fish oil on murine hepatic and hippocampal gene expression.

BACKGROUND: The functions, actions, and regulation of tissue metabolism affected by the consumption of long chain polyunsaturated fatty acids (LC-PUFA) from fish oil and other sources remain poorly understood; particularly how LC-PUFAs affect transcription of genes involved in regulating metabolism. In the present work, mice were fed diets containing fish oil rich in eicosapentaenoic acid and docosahexaenoic acid, fungal oil rich in arachidonic acid, or the combination of both. Liver and hippocampus tissue were then analyzed through a combined gene expression- and lipid- profiling strategy in order to annotate the molecular functions and targets of dietary LC-PUFA. RESULTS: Using microarray technology, 329 and 356 dietary regulated transcripts were identified in the liver and hippocampus, respectively. All genes selected as differentially expressed were grouped by expression patterns through a combined k-means/hierarchical clustering approach, and annotated using gene ontology classifications. In the liver, groups of genes were linked to the transcription factors PPARalpha, HNFalpha, and SREBP-1; transcription factors known to control lipid metabolism. The pattern of differentially regulated genes, further supported with quantitative lipid profiling, suggested that the experimental diets increased hepatic beta-oxidation and gluconeogenesis while decreasing fatty acid synthesis. Lastly, novel hippocampal gene changes were identified. CONCLUSIONS: Examining the broad transcriptional effects of LC-PUFAs confirmed previously identified PUFA-mediated gene expression changes and identified novel gene targets. Gene expression profiling displayed a complex and diverse gene pattern underlying the biological response to dietary LC-PUFAs. The results of the studied dietary changes highlighted broad-spectrum effects on the major eukaryotic lipid metabolism transcription factors. Further focused studies, stemming from such transcriptomic data, will need to dissect the transcription factor signaling pathways to fully explain how fish oils and arachidonic acid achieve their specific effects on health.

Journal Article↗

Evening primrose oil and fish oil in non-insulin-dependent-diabetes.

The effects of two anti-thrombotic and anti-lipidemic oils, evening primrose oil and fish oil, on glucose and lipid metabolism, prostaglandin (PG) levels and body composition were studied in patients with non-insulin-dependent diabetes. Seven patients were administered 4 g evening primrose oil, 2.4 g sardine oil and 200 mg vitamin E for 4 weeks. Fasting plasma glucose, hemoglobin A1c, total cholesterol, body weight and % body fat mass were significantly decreased after the treatment, and levels of changes in these parameters were not different from 11 patients who did not receive the oils. In the treatment group, concentrations of (e) icosapentaenoic acid (EPA) increased significantly in all the lipoprotein fractions, but dihomo-gamma-linolenic acid (DGLA) increased only in the high-density lipoprotein (HDL) fraction. The treatment decreased urinary 11-dehydro-thromboxane B2 excretion (32.7% decrease, P < 0.05), but did not alter significantly plasma PGE1 or 6-keto-PGF1 alpha levels. The ratio of 6-keto-PGF1 alpha and PGE1 to 11-dehydro-thromboxane B2 increased significantly after the treatment. These results suggest that these oil treatments are useful in improving abnormal lipid and thromboxane (TX)A2 metabolism in diabetic patients.

Blood Glucose↗

A double-blind comparison of olive oil and a combination of evening primrose oil and fish oil in the management of canine atopy.

A randomised double-blind parallel study lasting eight weeks was used to assess the effects of olive oil in a group of atopic dogs whose clinical signs were well controlled by dietary supplementation with a combination of evening primrose oil and fish oil. Nine of the 11 dogs which continued to receive this combination were considered unchanged at the conclusion of the study, whereas eight of the 10 dogs switched to olive oil had deteriorated. The mean plasma concentration of dihomogammalinolenic acid, a precursor of potentially antiinflammatory mediators, was significantly reduced (P < 0.05) in the olive oil-treated group at the end of the study. There were no significant differences between the mean plasma linoleic, eicosapentaenoic and arachidonic acid concentrations in the two groups. These findings suggest that olive oil is not an effective therapeutic agent in the control of canine atopy.

Animal Feed↗

A blinded, placebo-controlled study of the efficacy of borage seed oil and fish oil in the management of canine atopy.

Twenty-one dogs with atopy were entered into a blinded, placebo-controlled study lasting eight weeks. They were randomly divided into three groups and were all given supplementary oils orally once daily. The dogs in groups A and B were given borage seed oil and fish oil in combination (Viacutan; Boehringer Ingelheim Vetmedica) to provide 176 mg/kg or 88 mg/kg borage seed oil respectively. The dogs in group C were given 204 mg/kg olive oil as a placebo. They were all re-examined after four and eight weeks and scored for pruritus, erythema, oedema, alopecia and self-excoriation. After eight weeks the scores for erythema and self-excoriation, and the total score for the dogs in group A, and the total score for the dogs in group B were significantly reduced (P < 0.05). The dogs in group C showed no significant improvement.

Animals↗

The effect on human tumor necrosis factor alpha and interleukin 1 beta production of diets enriched in n-3 fatty acids from vegetable oil or fish oil.

The effect of a flaxseed oil-based diet on tumor necrosis factor alpha (TNF alpha) and interleukin 1 beta (IL-1 beta) synthesis was examined in healthy volunteers. Use of flaxseed oil in domestic food preparation for 4 wk inhibited TNF alpha and IL-1 beta production by approximately 30%. Fish-oil supplementation (9 g/d) continued for a further 4 wk; TNF alpha and IL-1 beta synthesis were inhibited by 74% and 80%, respectively. There was a significant inverse exponential relation between TNF alpha or IL-1 beta synthesis and mononuclear cell content of eicosapentaenoic acid (EPA), an n--3 fatty acid derived from ingested EPA (fish oil) or metabolism of ingested alpha-linolenic acid (flaxseed oil). Cytokine production decreased as cellular EPA increased to approximately 1% of total fatty acids. Further increases in EPA content did not result in further decreases in cytokine production. The results indicate that vegetable oils rich in n--3 fatty acids inhibit TNF alpha and IL-1 beta synthesis.

Adult↗

Increased lipid peroxidation and impaired antioxidant enzyme function is associated with pathological liver injury in experimental alcoholic liver disease in rats fed diets high in corn oil and fish oil.

Increased hepatic oxidative stress with ethanol administration is hypothesized to be caused either by enhanced pro-oxidant production or decreased levels of antioxidants or both. We used the intragastric feeding rat model to assess the relationship between hepatic antioxidant enzymes and pathological liver injury in animals fed different dietary fats. Male Wistar rats (5 per group) were fed ethanol with either medium-chain triglycerides (MCTE), palm oil (PE), corn oil (CE), or fish oil (FE). Control animals were fed isocaloric amounts of dextrose instead of ethanol with the same diets. The following were evaluated in each group: liver pathology, lipid peroxidation, manganese superoxide dismutase (MnSOD) levels, copper-zinc SOD (CuZnSOD) levels, glutathione peroxidase (GPX) levels, and catalase (CAT) levels. All enzymes were evaluated using activity assays and immunoblots. Rats fed FE showed the most severe pathology (fatty liver, necrosis, and inflammation), those fed CE showed moderate changes, those fed PE showed fatty liver only, and those fed MCTE were normal. Parameters indicative of lipid peroxidation (conjugated dienes and thiobarbituric acid-reactive substances) were also greater in rat livers from animals fed the diets high in polyunsaturated fatty acids (CE and FE). CuZnSOD, GPX, and CAT activities showed an inverse correlation (r=-.92, P < .01) with severity of pathological injury, with the lowest levels for both enzymes found in FE-fed rats. Decreased enzyme activity in CE- and FE-fed rats was accompanied by similar decreases in immunoreactive protein. Ethanol administration did not cause significant decreases in enzyme activity in groups that showed no necroinflammatory changes (MCTE and PE). MnSOD activity showed no significant change in any ethanol-fed group. Our results show that decreases in CuZnSOD, GPX, and CAT occur in rats showing pathological liver injury and also having the highest levels of lipid peroxidation. These results suggest that feeding dietary substrates that enhance lipid peroxidation can exacerbate both ethanol-induced oxidative damage as well as necroinflammatory changes. The decrease in activity of antioxidant enzymes observed in animals fed diets high in polyunsaturated fatty acids and ethanol could possibly increase the susceptibility to oxidative damage and further contribute to ethanol-induced liver injury.

Animals↗

Influence of formulas with borage oil or borage oil plus fish oil on the arachidonic acid status in premature infants.

Several studies have reported that feeding gamma-linolenic acid (GLA) has resulted in no increase in arachidonic acid (AA) in newborns. This result was ascribed to the eicosapentaenoic acid (EPA)-rich fish oil used in these formulas. Docosahexaenoic acid (DHA) sources with only minor amounts of EPA are now available, thus the addition of GLA to infant formulas might be considered an alternative to AA supplementation. Sixty-six premature infants were randomized to feeding one of four formulas [ST: no GLA, no long-chain polyunsaturated fatty acids; BO: 0.6% GLA (borage oil); BO + FOLOW: 0.6% GLA, 0.3% DHA, 0.06% EPA; BO + FOHIGH: 0.6% GLA, 0.3% DHA, 0.2% EPA] or human milk (HM, nonrandomized) for 4 wk. Anthropometric measures and blood samples were obtained at study entry and after 14 and 28 d. There were no significant differences between groups in anthropometric measures, tocopherol, and retinol status at any of the studied time points. The AA content of plasma phospholipids was similar between groups at study start and decreased significantly until day 28 in all formulafed groups, but not in the breast-fed infants [ST: 6.6 +/- 0.2%, BO: 6.9 +/- 0.3%, BO + FOLOW: 6.9 +/- 0.4%, BO + FOHIGH: 6.7 +/- 0.2%, HM: 8.6 +/- 0.5%, where values are reported as mean +/- standard error; all formulas significantly different (P< 0.05) from HM]. There was no significant influence of GLA or fish oil addition to the diet. GLA had only a very limited effect on AA status which was too small to obtain satisfactory concentrations (concentrations similar to breast-fed babies) under the circumstances tested. The effect of GLA on AA is independent of the EPA and DHA content in the diet within the dose ranges studied.

Antioxidants↗

Melanoma lung metastases and cytolytic effector cells in mice fed antioxidant-balanced corn oil or fish oil diets.

(C57BL/6 x DBA/2)F1 mice were fed antioxidant-matched fish oil (FO) or corn oil (CO) diets for weeks, were challenged with B16.F10 melanoma cells, and lung metastases were enumerated 17 days later. Mice fed FO had fewer lung tumors than mice fed CO. This dietary effect persisted in mice injected with monoclonal antibodies which depleted natural killer cells, CD8+ T cells or CD4+ T cells. Generation of specific anti-B16.F10 cytotoxic cells in vitro by splenocytes from immunized mice was greater in mice fed FO. Even though host resistance to lung metastases by B16.F10 cells is mediated in large part by natural killer cells, CO and/or FO may have affected cells not tested here or tumor cells themselves.

Animals↗

Effect of soybean oil and fish oil on individual molecular species of Atlantic salmon head kidney phospholipids determined by normal-phase liquid chromatography coupled to negative ion electrospray tandem mass spectrometry.

The effect of soybean oil (SO) and fish oil (FO) on the relative molecular species distribution of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI) and phosphatidylserine (PS) in Atlantic salmon head kidney was studied using normal-phase liquid chromatography coupled with negative ion electrospray tandem mass spectrometry. The conformation of identity of the phospholipid species was based on retention time, the mass of the [M-H]- ([M-15]- for PC) molecular ions and the carboxylate anion fragments in the product ion spectrum. The intensity ratio of sn-1/sn-2 fragment ions increased with increasing number of double bonds in the sn-2 acyl chain but was not affected by increasing number of double bonds in the sn-1 acyl chain of the species examined. The relative distribution of the molecular species was determined by multiple reaction monitoring of the carboxylate anion fragment from the sn-1 position. A total of 68 different phospholipid species were determined in the head kidney and the largest amount was found in PE (22 species). Depending on the diet, the main species identified in the different phospholipid classes were; PC 16:0/18:1, PE 16:0/22:6, PI 18:0/20:4 and PS 16:0/22:6. The SO diet significantly increased the 18:2, 20:3 and most 20:4 containing species and significantly reduced the 14:0 and most 20:5 and 22:6 fatty acid containing species. The increase of the 20:4 and the decrease of the 20:5 and 22:6 containing species were dependent on the fatty acid combination of the species.

Animals↗

Dietary manipulation by perilla oil and fish oil of hepatic lipids and its influence on peroxisomal beta-oxidation and serum lipids in rat and mouse.

Rats and mice were fed a diet, prepared with soybean oil (SO), perilla oil (PO) or fish oil (FO), for 4 weeks. Compared with the groups of SO-feeding, FO-feeding increased content of eicosapentaenoic acid (20:5 n-3), docosapentaenoic acid (22:5 n-3) and docosahexaenoic acid (22:6 n-3) and PO-feeding elevated the content of alpha-linolenic acid (18:3 n-3), 20:5 n-3 and 22:5 n-3 or hepatic lipids of both rats and mice. FO-feeding increased the activity of peroxisomal beta-oxidation in the livers of both rats and mice. The activities of peroxisomal beta-oxidation in rats and mice that were fed FO diet at a high fat (40% of energy) concentration corresponded to 20% and 30%, respectively, of the maximum activities induced by peroxisome proliferators (4-chlorophenoxyisobutyric acid or perfluorooctanoic acid). To a lesser extent, PO-feeding elevated this activity too. There were significant correlations between peroxisomal beta-oxidation activity and the content of either 20:5 n-3, 22:5 n-3 or 22:6 n-3 in hepatic lipids. FO-feeding decreased effectively serum level of cholesterol of both rats and mice. The reduction in serum cholesterol by feeding PO was less pronounced than that observed with FO-feeding. A high correlation was found between 22:6 n-3 content in hepatic lipids and serum concentration of cholesterol. Although FO-feeding lowered the level of circulating triacylglycerol, PO-feeding produced no change. No substantial correlation was observed between the hepatic content of n-3 fatty acid and the concentration of serum triacylglycerol.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

The combined effects of garlic oil and fish oil on the hepatic antioxidant and drug-metabolizing enzymes of rats.

This present study was designed to investigate the combined modulatory effect of garlic oil (GO) and fish oil (FO) on the antioxidant and drug metabolism systems. Rats were fed either a low-maize oil (MO) diet (50 g MO/kg), high-MO diet (235 g MO/kg) or high-FO diet (205 g FO+ 30 g MO/kg) and received different doses of GO (0-200 mg/kg body weight) three times per week for 6 weeks. Fatty acid analysis showed that 20 : 5n-3 and 22 : 6n-3 were incorporated into serum lipid at the expense of 18 : 2n-6 and 20 : 4n-6 in rats fed the high-FO diet. GO dose-dependently increased hepatic glutathione S-transferase (GST), glutathione reductase, superoxide dismutase (SOD) and ethoxyresorufin O-deethylase (EROD) activities, but decreased glutathione peroxidase and N-nitrosodimethylamine demethylase (NDMAD) activities (P<0.05). With the exception of glutathione peroxidase, the activities of glutathione reductase, SOD, GST, EROD and NDMAD were modulated by the dietary fat. The high-FO group had greater SOD and EROD activity than either MO-fed group; it also had greater NDMAD activity than the low-MO group (P<0.05). GST activity was higher in rats fed high-FO or high-MO diets than rats fed the low-MO diet. Change in erythromycin demethylase activity, however, was not caused by either dietary fat or GO. Immunoblot assay showed that GO dose-dependently enhanced the protein level of the Ya, Yb1, Yc isoenzymes of GST and cytochrome P450 (CYP) 1A1 and 3A1, but GO suppressed CYP2E1 expression. Regardless of the dosage of GO, the high-FO diet increased CYP1A1, CYP3A1 and CYP2E1 levels compared with the high- and low-MO diets. Accompanying the changes observed in immunoblots, CYP1A1 and CYP3A1 mRNA levels were increased by GO in a dose-dependent manner and also increased additively in combination with FO feeding. These present results indicate that co-administration of GO and FO modulates the antioxidant and drug-metabolizing capacity of animals and that the effect of GO and FO on drug-metabolizing enzymes is additive.

Allyl Compounds↗

Effect of dietary intake of fish oil and fish protein on the development of L-azaserine-induced preneoplastic lesions in the rat pancreas.

The effect of dietary intake of fish (menhaden) oil and fish (cod) protein on the development of pancreatic preneoplastic lesions was examined in male Wistar rats. Fourteen-day-old animals were given a single ip injection of 30 mg L-azaserine/kg body weight [CAS: 115-02-6; diazoacetate serine (ester)]. At 21 days of age they were weaned and maintained on dietary treatment for 4 months. Fish protein did not appear to produce a significantly different preneoplastic response when compared to casein as a protein source. However, a 20% menhaden oil diet, rich in omega 3 fatty acids, produced a significant decrease in the development of both the size and number of preneoplastic lesions when compared to a 20% corn oil diet rich in omega 6 fatty acids. This study provides evidence that fish oils, rich in omega 3 fatty acids, may have potential as inhibitory agents in cancer development.

Animals↗

Effect of high fat corn oil, olive oil and fish oil on phospholipid fatty acid composition in male F344 rats.

Epidemiological and laboratory animal model studies have provided evidence that the effect of dietary fat on colon tumorigenesis depends on the amount of fat and its composition. Because of the importance of the composition of dietary fat and of tissue membrane fatty acid composition in tumor promotion, experiments were designed to investigate the relative effects of high fat diets rich in omega 3, omega 6 and omega 9 fatty acids and colon carcinogen on the phospholipid fatty acid composition of liver, colon, small intestine, erythrocytes and blood plasma. At 6 wk of age, groups of animals were fed diets containing 5% corn oil (LFCO), 23.5% corn oil (HFCO), 23.5% olive oil (HFOO), and 20.5% fish oil plus 3% corn oil (HFFO). Two weeks later all the animals except the vehicle-treated animals received azoxymethane s.c. once weekly for 2 wk at a dose rate of 15 mg/kg body weight. Animals were sacrificed 5 d later and liver, colon, small intestine and erythrocytes and blood plasma were analyzed for phospholipid fatty acids. The results indicate that the phospholipid fatty acid composition of liver, colon and small intestine of HFCO diet fed animals, were not significantly different from those fed the LFCO diet. The levels of palmitoleic acid and linoleic acid were increased in erythrocytes and blood plasma of the animals fed the HFCO diet compared to those fed the LFCO diet. Feeding the HFOO diet significantly increased the oleic acid content and decreased the linoleic acid and arachidonic acid levels in various organs when compared to the HFCO diet.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Dietary marine fatty acids (fish oil) for asthma.

BACKGROUND: Epidemiological studies suggest that a diet high in marine fatty acids (fish oil) may have beneficial effects on inflammatory conditions such as rheumatoid arthritis and possibly asthma. OBJECTIVES: 1. To determine the effect of marine n-3 fatty acid (fish oil) supplementation in asthma. 2. To determine the effect of a diet high in fish oil in asthma. SEARCH STRATEGY: The Cochrane Airways Review Group register was search using the terms: marine fatty acids OR diet OR nutrition OR fish oil OR eicosapentaenoic acid OR EPA. Bibliographies of retrieved trials were searched and fish oil manufacturers contacted. SELECTION CRITERIA: Randomised controlled trials in patients with asthma more than two years of age were included. The study duration had to be in excess of 4 weeks. Double blind trials were preferred, but single-blind and open trials were also reviewed for possible inclusion. Three reviewers read each paper, blind to its identity. Decisions concerning inclusion were made by simple majority. Quality assessment was performed by all three reviewers independently. DATA COLLECTION AND ANALYSIS: The only comparison possible was between marine n-3 fatty acid supplementation and placebo. There were insufficient trials to examine dietary manipulation alone. MAIN RESULTS: Eight randomised controlled trials conducted between 1986 and 1998 satisfied the inclusion criteria. Six were of parallel design and two were cross-over studies. Seven compared fish oil with placebo whilst one compared high dose vs low dose marine n-3 fatty acid supplementation. None of the included studies reported asthma exacerbations, health status or hospital admissions. There was no consistent effect on any of the analyzable outcomes: FEV1, peak flow rate, asthma symptoms, asthma medication use or bronchial hyper reactivity. The single study performed in children also combined dietary manipulation with fish oil supplementation and showed improved peak flow and reduced asthma medication use. There were no adverse events associated with fish oil supplements. REVIEWER'S CONCLUSIONS: There is little evidence to recommend that people with asthma supplement or modify their dietary intake of marine n-3 fatty acids (fish oil) in order to improve their asthma control. Equally, there is no evidence that they are at risk if they do so.

Adult↗

Dietary supplementation with primrose oil or fish oil does not change urinary excretion of prostacyclin and thromboxane metabolites in pre-eclamptic women.

To study whether balance between antiaggregatory, vasodilatory prostacyclin (PGI2) and proaggregatory, vasoconstrictory thromboxane A2 (TXA2) could be affected by dietary manipulation, 18 pre-eclamptic women were treated in randomized order between 31 and 36 weeks of gestation either with primrose oil (n = 7), with fish oil (n = 5), or with placebo (n = 6). Urinary excretions of the degradation products of PGI2 (6-keto-PGF1 alpha, 2,3-dinor-6-keto-PGF1 alpha) and TXA2 (TXB2, 2,3-dinor-TXB2) were measured in 24 h urines before and serially during the supplementation. Fatty acid supplementation did not affect urinary prostanoid excretions or clinical signs of pre-eclampsia.

Epoprostenol↗