Rats Fed Fish Oil Research Articles

Dietary Medium-Chain Fatty Acids Raise and (n-3) Polyunsaturated Fatty Acids Lower Hepatic Triacylglycerol Synthesis in Rats

The hypothesis tested was that dietary medium-chain or (n-3) polyunsaturated fatty acids, when compared with (n-6) polyunsaturated fatty acids, alter plasma triacylglycerol levels by affecting hepatic triacylglycerol synthesis as reflected by the activities of acetyl-CoA carboxylase, fatty acid synthase and diacylglycerol acyltransferase in liver. In two separate experiments rats were fed purified diets containing (n-6) polyunsaturated fatty acids in the form of corn oil and either (n-3) polyunsaturated fatty acids in the form of fish oil or medium-chain triacylglycerols (MCT). Consumption of MCT significantly raised plasma triacylglycerol concentrations, whereas fish oil feeding had a lowering effect compared with the corn oil-fed group. In individual rats, the hepatic triacylglycerol concentration was directly correlated with the plasma triacylglycerol concentration (r = 0.60, P < 0.001). The MCT oil diet vs. the corn oil diet markedly raised the activities of hepatic acetyl-CoA carboxylase, fatty acid synthase and diacylglycerol acyltransferase. In the rats fed fish oil, the activities of fatty acid synthase and diacylglycerol acyltransferase were significantly reduced, whereas the activity of acetyl-CoA carboxylase was not affected relative to activities in rats fed corn oil. The activities of the three enzymes were directly correlated with plasma triacylglycerol concentrations in individual rats (r = 0.60–0.75, P < 0.001). The type of fat in the diet probably affects the rate of hepatic triacylglycerol synthesis which is an important determinant of plasma triacylglycerol concentrations.

Fish Oil Enhances Pentachlorobenzene Metabolism and Reduces its Accumulation in Rats

To investigate the influence of dietary fats on the metabolism and excretion of pentachlorobenzene (PECB), rats were fed semipurified diets containing soybean oil, lard or fish oil (10 g/100 g diet) for 2 wk. Rats then received a single dose of PECB by intragastric gavage. Blood concentration of pentachlorophenol (PCP), a major metabolite of PECB, was higher after 5 h through d 3 and that of PECB was lower after d 5 in the rats fed fish oil than in rats fed the other fats. After d 5, the highest ratio of PCP/PECB in the liver was seen in the fish oil—fed rats. Furthermore, fish oil intake resulted in a markedly lower concentration of PECB in fat tissues, where a large amount of PECB is typically distributed due to its lipophilic property. These findings indicate that fish oil feeding enhances PECB metabolism, thereby decreasing PECB residues in the body. Fish oil—fed rats had a smaller epididymal fat tissues mass compared with lard- or soybean oil—fed rats. The small mass of the fat tissues apparently limited the accumulation of PECB in those tissues and thus accelerated PECB metabolism in the liver.

Dietary Fish Oil Supplementation Attenuates Myocardial Dysfunction and Injury Caused by Global Ischemia and Reperfusion in Isolated Rat Hearts

To determine the effect of dietary fish oil supplementation on myocardial dysfunction following ischemia-reperfusion, independent of plasma and blood cells, Sprague-Dawley rats were fed fish oil-rich nonpurified diet or butter-enriched diet for 5 d. Myocardial content of long-chain and (n-3) polyunsaturated fatty acids was greater in the fish oil-fed rats (P < 0.01), whereas (n-6) fatty acid content was lower compared with controls (P < 0.01). Platelet aggregation in fish oil-fed rats was also inhibited. Hearts from all rats were subjected to 15 min of global ischemia and 10 min of reperfusion. In hearts of control rats, ischemia-reperfusion resulted in a marked decrease in force of cardiac contraction, increase in coronary perfusion pressure, appearance of ventricular arrhythmias and release of creatine kinase and thromboxane B2 in the coronary effluent. Dietary fish oil supplementation attenuated myocardial dysfunction induced by ischemia-reperfusion, as indicated by smaller change in force of cardiac contraction (-77% vs. -89%, P < 0.05) and coronary perfusion pressure (+37% vs. +71%, P < 0.001). Concomitantly, release of creatine kinase as well as thromboxane B2 in coronary effluent was lower (P < 0.01). Ventricular arrhythmias occurred less frequently in hearts from fish oil-fed rats. Thus, short-term dietary fish oil supplementation attenuates myocardial dysfunction caused by ischemia-reperfusion by a direct effect on the heart independent of blood and plasma components.

Dietary Fish Oil Does Not Alter Glucose Tolerance in Conscious Rats

We examined the effect of dietary fish oil (MaxEPA) and sunflower seed oil on glucose tolerance in male Wistar rats. Semipurified diets containing 100 g oil/kg diet were administered for 30 d. The fish oil diet contained 26 g (n-3) fatty acids, 16 g eicosapentaenoic acid and 10.4 g docosahexaenoic acid/kg diet. Phospholipids from liver, pancreas, and pancreatic islets were enriched in eicosapentaenoic and docosahexaenoic acids by the fish oil diet. In unfed pentobarbital-anesthetized rats, both basal plasma insulin concentration and insulin responses to intravenous glucose were significantly lower for fish oil-fed rats although glucose responses were similar; however, incremental excursions in plasma insulin over the basal concentrations did not differ. Intravenous glucose tolerance was also examined in conscious unfed rats under minimal restraint. Responses of plasma glucose and insulin were similar for fish oil- and sunflower oil-fed groups. Furthermore, in another experiment, intravenous glucose tolerance tests were similar for conscious rats provided with either 100 g fish oil or corn oil/kg nonpurified diet. Thus, glucose-induced insulin secretion is lower in rats fed fish oil than in rats fed sunflower oil, when tests are conducted in pentobarbital-anesthetized animals but not when tests are performed in conscious rats; there was no effect on plasma glucose in either anesthetized or nonanesthetized rats. Therefore, substitution of (n-3) for (n-6) polyunsaturated fatty acids in tissue phospholipids does not alter plasma glucose or insulin in conscious male Wistar rats.

Dietary Fatty Acid Composition Influences Energy Accretion in Rats

To study the influence of dietary fatty acid composition on energy metabolism, forty male rats were fed elemental diets containing 42% of energy as fish oil, safflower oil, olive oil or beef tallow for 12 wk. Food intakes and body weights were measured daily. Energy expenditure and body composition were determined using doubly labeled water on the final 2 d. Pooled fecal energy losses differed in response to dietary manipulation, with losses greatest in rats fed beef tallow and least in those fed olive oil. Higher lean body mass gains and lower fat mass gains were observed in rats fed diets containing fish oil, compared with rats fed olive oil or beef tallow. Total energy gains in rats fed olive oil (3632 +/- 145 kJ) and beef tallow (3850 +/- 136 kJ) were higher than those in rats fed fish oil (2905 +/- 196 kJ). Energy efficiency in both the olive oil and beef tallow groups was also higher than that in the fish oil group. There were no differences in energy expenditure measured by doubly labeled water technique among the four groups. The data suggest that dietary fatty acid composition alters the efficiency of energy substrate accretion in rats.

Differential Effect of Four-Week Feeding of Different Dietary Fats on the Accumulation of Fat and the Cholesterol and Triglyceride Contents in the Different Fat Depots.

The aim of the present study was to determine the effects of feeding of a high-fat diet containing different types of lipids for four weeks on the cholesterol and triglyceride contents of different fat depots and on body temperature in rats. Four groups of adult rats were fed 10% fat, containing either beef tallow, safflower oil, or fish oil, respectively, as well as a normal rodent diet with 4% fat, for four weeks. The rats on normal rodent diet consumed significantly more food and water than the rats in the other three groups. Rectal temperature increased only after four-week feeding with safflower oil fat. Increased fat deposition and adipocyte size were observed in rats fed safflower oil and beef tallow. In all fat pads of safflower oil-fed rats, cholesterol content was significantly higher than the other three groups. Feeding of beef tallow increased triglyceride depot without increasing cholesterol content. The rats fed fish oil had significantly less triglyceride and cholesterol deposition in adipose tissues than the rats fed safflower oil or beef tallow. These results clearly demonstrated the differences in fat deposition, adipocyte size and number, triglyceride and cholesterol accumulation in fat cells are dependent on the dietary lipid composition.

Dietary polyunsaturated fatty acids modulate adipose secretome and is associated with changes in mammary epithelial stem cell self-renewal

Chronic low-grade adipose inflammation, characterized by aberrant adipokine production and pro-inflammatory macrophage activation/polarization is associated with increased risk of breast cancer. Adipocyte fatty acid composition is influenced by dietary availability and may regulate adipokine secretion and adipose inflammation. After feeding F344 rats for 20 weeks with a Western diet or a fish oil-supplemented diet, we cultured primary rat adipose tissue in a three-dimensional explant culture and collected the conditioned medium. The rat adipose tissue secretome was assayed using the Proteome Profiler Cytokine XL Array, and adipose tissue macrophage polarization (M1/M2 ratio) was assessed using the iNOS/ARG1 ratio. We then assessed the adipokine's effects upon stem cell self-renewal using primary human mammospheres from normal breast mammoplasty tissue. Adipose from rats fed the fish oil diet had an ω-3:ω-6 fatty acid ratio of 0.28 compared to 0.04 in Western diet rats. The adipokine profile from the fish oil-fed rats was shifted toward adipokines associated with reduced inflammation compared to the rats fed the Western diet. The M1/M2 macrophage ratio decreased by 50% in adipose of fish oil-fed rats compared to that from rats fed the Western diet. Conditioned media from rats fed the high ω-6 Western diet increased stem cell self-renewal by 62%±9% (X¯%±SD) above baseline compared to only an 11%±11% increase with the fish oil rat adipose. Modulating the adipokine secretome with dietary interventions therefore may alter stromal–epithelial signaling that plays a role in controlling mammary stem cell self-renewal.

Dietary Fat Influences the Expression of Contractile and Metabolic Genes in Rat Skeletal Muscle

Dietary fat plays a major role in obesity, lipid metabolism, and cardiovascular diseases. To determine whether the intake of different types of dietary fats affect the muscle fiber types that govern the metabolic and contractile properties of the skeletal muscle, we fed male Wistar rats with a 15% fat diet derived from different fat sources. Diets composed of soybean oil (n-6 polyunsaturated fatty acids (PUFA)-rich), fish oil (n-3 PUFA-rich), or lard (low in PUFAs) were administered to the rats for 4 weeks. Myosin heavy chain (MyHC) isoforms were used as biomarkers to delineate the skeletal muscle fiber types. Compared with soybean oil intake, fish oil intake showed significantly lower levels of the fast-type MyHC2B and higher levels of the intermediate-type MyHC2X composition in the extensor digitorum longus (EDL) muscle, which is a fast-type dominant muscle. Concomitantly, MyHC2X mRNA levels in fish oil-fed rats were significantly higher than those observed in the soybean oil-fed rats. The MyHC isoform composition in the lard-fed rats was an intermediate between that of the fish oil and soybean oil-fed rats. Mitochondrial uncoupling protein 3, pyruvate dehydrogenase kinase 4, and porin mRNA showed significantly upregulated levels in the EDL of fish oil-fed rats compared to those observed in soybean oil-fed and lard-fed rats, implying an activation of oxidative metabolism. In contrast, no changes in the composition of MyHC isoforms was observed in the soleus muscle, which is a slow-type dominant muscle. Fatty acid composition in the serum and the muscle was significantly influenced by the type of dietary fat consumed. In conclusion, dietary fat affects the expression of genes related to the contractile and metabolic properties in the fast-type dominant skeletal muscle, where the activation of oxidative metabolism is more pronounced after fish oil intake than that after soybean oil intake.

Effects of contaminated fish oil on aberrant crypt foci formation and colonic cell kinetics

Fish oil is a rich source of n‐3 polyunsaturated fatty acids, which are known to prevent colon cancer. However this message is confounded by Food Drug Administration warnings which state that consumption of certain types of fish should be restricted due to contaminants such as polychlorinated biphenyls (PCBs). Therefore, this study examined the effects of fish oil contaminated with PCB (4000ng/ml fat) as compared to regular fish oil on the risk factors of colon cancer including aberrant crypt foci (ACF) formation and colonic cell kinetics (cell proliferation and apoptosis). Thirty Sprague‐Dawley rats were divided into three groups and fed corn oil, fish oil, or contaminated fish oil (PCB) for nine weeks and injected with saline or azoxymethane (2 times at wk 3 and 4). Fish oil‐fed rats had a lowered number of ACF (P=0.013) and high multiplicity ACF (P=0.024) compared to the corn oil and contaminated fish oil groups. Fish oil feeding resulted in lower cell proliferation index than corn oil and PCB groups (P=0.009). Cell apoptosis was not significantly different amongst any of the groups. These results suggest that dietary fish oil protects against colon cancer by lowering the number of early preneoplastic lesions through the inhibition of cell proliferation. PCB contaminated fish oil reduced the beneficial effects of fish oil on colon carcinogenesis. Supported by CHNR08‐810, SDSU UGP and NUTR 302L class.

Trans Fatty Acids: Chemical Synthesis of Eicosapentaenoic Acid Isomers and Detection in Rats Fed a Deodorized Fish Oil Diet

Eicosapentaenoic acid (EPA) is a polyunsaturated fatty acid present in fish oils used for omega-3 enriched diets. The natural cis double bond geometry can be transformed to the trans configuration during the deodorization process utilized in the food industry. The analytical discrimination of the possible five monotrans regioisomers represents a limiting step for the recognition and structure-activity relationship in connection with the harmful effects of trans fatty acids in health. We carried out a dual synthetic strategy, providing new access to monotrans EPA isomers and valuable information on GC and NMR characteristics for further applications in metabolomics and lipidomics. This small library was used as an analytical reference for isomer determination in deodorized fish oils and the follow-up of rats fed fish oil diets, evidencing for the first time that monotrans EPA isomers are incorporated in liver mitochondrial membranes after dietary intake.

Effect of fish oil and coconut fat supplementation on depressive-type behavior and corticosterone levels of prenatally stressed male rats

Prenatal stress (PNS) during critical periods of brain development has been associated with numerous behavioral and/or mood disorders in later life. These outcomes may result from changes in the hypothalamic-pituitary-adrenal (HPA) axis activity, which, in turn, can be modulated by environmental factors, such as nutritional status. In this study, the adult male offspring of dams exposed to restraint stress during the last semester of pregnancy and fed different diets were evaluated for depressive-like behavior in the forced swimming test and for the corticosterone response to the test. Female Wistar rats were allocated to one of three groups: regular diet, diet supplemented with coconut fat or with fish oil, offered during pregnancy and lactation. When pregnancy was confirmed, they were distributed into control or stress groups. Stress consisted of restraint and bright light for 45min, three times per day, in the last week of pregnancy. The body weight of the adult offspring submitted to PNS was lower than that of controls. In the forced swimming test, time of immobility was reduced and swimming was increased in PNS rats fed fish oil and plasma corticosterone levels immediately after the forced swimming test were lower in PNS rats fed regular diet than their control counterparts; this response was reduced in control rats whose mothers were fed fish oil and coconut fat. The present results indicate that coconut fat and fish oil influenced behavioral and hormonal responses to the forced swimming test in both control and PNS adult male rats.

Dietary fatty acids and oxidative stress in the heart mitochondria

Our study compared the effects of different oils on oxidative stress in rat heart mitochondria, as well as on plasma parameters used as risk factors for cardiovascular disease. The rats were fed for 16 weeks with coconut, olive, or fish oil diet (saturated, monounsaturated, or polyunsaturated fatty acids, respectively). The cardiac mitochondria from rats fed with coconut oil showed the lowest concentration of oxidized proteins and peroxidized lipids. The fish oil diet leads to the highest oxidative stress in cardiac mitochondria, an effect that could be partly prevented by the antioxidant probucol. Total and LDL cholesterols decreased in plasma of rats fed fish oil, compared to olive and coconut oils fed rats. A diet enriched in saturated fatty acids offers strong advantages for the protection against oxidative stress in heart mitochondria.

Dietary fish oil impairs induction of γ‐interferon and delayed‐type hypersensitivity during a systemic Salmonella enteritidis infection in rats

Fish oil that is rich in n-3 polyunsaturated fatty acids markedly modulates immunological responses. Literature data indicate that the fish oil reduces cellular immunity and therefore impairs resistance to infections. We have investigated how dietary fish oil affects the immune response against a facultative intracellular pathogen, Salmonella enteritidis. Wistar rats were fed a diet containing 16% (w/w) of either fish oil or corn oil. After a 4-week adaptation period, rats were intraperitoneally challenged with 4 x 10(5) cfu of S. enteritidis. During the 14-day infection period, urine was collected on a daily basis. At days 2 and 14, eight rats per group were sacrificed. Urinary nitrate, used as a marker for NO production, was lower on a fish oil diet during days 3-8. At day 2, serum gamma-interferon was 48 +/- 7 pg/mL in the fish oil-fed rats compared with 162 +/- 52 pg/mL in the corn oil-fed rats. No effects were found on living salmonella in liver and spleen. At day 14, as markers of an impaired T-helper 1 (Th-1) response, a 38% lower delayed-type hypersensitivity responses and a lower salmonella-specific IgG2b were observed in the fish oil-fed rats. Although here dietary fish oil has affected only immune parameters, this impairment of the innate and Th-1-mediated immune response may have implications for the host resistance against other intracellular pathogens.

Dietary fatty acid composition alters magnesium metabolism, distribution, and marginal deficiency response in rats*

Based on dietary intake recommendations, magnesium deficiency commonly occurs throughout the world. However, widespread pathological conditions induced by dietary magnesium deficiency have not been identified. This discrepancy may be caused by other dietary factors ameliorating or exacerbating the response to a marginal magnesium deficiency and/or the length of the deficiency. Thus, a study was performed to determine whether the n-6/n-3 fatty acid composition of the diet affects the response to marginal magnesium deprivation, and whether the effect was dependent upon the length of deprivation. Weanling female rats were fed diets containing 250 mg/kg magnesium in a factorial arrangement with dietary variables of supplemental magnesium at 0 or 250 mg/kg (total of 250 or 500 mg/kg) and fat sources of 75 g/kg corn oil or 65 g/kg fish (menhaden) oil plus 10 g/kg linoleic acid. After 8 and 12 weeks on their respective diets, each rat was placed in a metabolic cage for a 16-hour collection of urine. After 13 weeks, the rats were anesthetized with ether for the collection of plasma and organs. Marginal magnesium deficiency was confirmed by decreased urinary excretion and femur, tibia and vertebrae concentrations of magnesium. Dietary oil influenced the effect of marginal magnesium deficiency on magnesium metabolism, distribution and oxidative stress indicators. Fish oil, but not corn oil, significantly decreased urinary magnesium excretion and increased kidney magnesium concentration. Femur magnesium was significantly decreased by marginal magnesium deficiency in rats fed fish oil but not in rats fed corn oil, and liver magnesium concentration was decreased by fish oil. Marginal magnesium deficiency increased plasma extracellular superoxide dismutase and cysteine (component of glutathione) in rats fed corn oil but not in rats fed fish oil. Urinary prostaglandin E₂ excretion was significantly decreased by marginal magnesium deficiency at 8 weeks, but not at 12 weeks; an increase between weeks 8 and 12 in marginally magnesium-deficient rats fed fish oil caused this change in significance. The findings show that the dietary fatty acid composition affects the response of rats to marginal magnesium deprivation. The findings also indicate that dietary or physiological factors affecting oxidative stress could affect the response to marginal magnesium deficiency, and that a response to a dietary change that takes time to develop, such as an increase in dietary n-3 fatty acids, may result in signs of marginal deficiency being different over time.

A Voluntary Oral Ethanol‐Feeding Rat Model Associated With Necroinflammatory Liver Injury

The intragastric (IG) ethanol infusion model results in fatty liver, necrosis, inflammation and fibrosis. This model was utilized to study the pathogenesis of alcoholic liver disease (ALD). Disadvantages of the IG model include maintenance of the animals and equipment expense. To develop a voluntary feeding model for ALD, we took advantage of two important observations in the IG model: (i) female rats demonstrate greater severity of alcohol-induced liver injury than males and (ii) rats fed fish oil as a source of fatty acids develop more severe alcoholic liver injury than rats fed other fatty acids with ethanol. Female Wistar rats (205 to 220 g) were fed for 8 weeks a diet containing 8% ethanol, fish oil (30% of calories), protein, and dextrose. Pair-fed controls (FD) received dextrose in amounts isocaloric to ethanol. The following measurements were made: liver pathology [fatty liver (0 to 4), necrosis, inflammation and fibrosis by Sirius Red], endotoxin and alanine aminotransferase (ALT) in plasma, urine ethanol, lipid peroxidation, nuclear factor kappa-B (NF-kappaB) and mRNA levels for tumor necrosis factor-alpha (TNF-alpha), cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS). Protein levels for iNOS and nitrotyrosine were evaluated by immunohistochemistry and Western Blot analysis. Liver proteasome and cytochrome P450 2E1 activity and protein levels of asialoglycoprotein receptor (ASGPR) were also evaluated. In addition, mRNA levels of fibrogenic markers were assessed. All animals lost weight for the initial 2 to 3 weeks but then gained weight until killing at 8 weeks. There was, however, a significant difference (p < 0.05) in weight between the ethanol-fed (Etoh) and (FD) groups at the end of the experiment. The mean urine ethanol levels ranged between 190 and 240 mg/dl. The severity of pathological changes was greater (p < 0.01) in Etoh vs. FD: fatty liver, 3.0 +/- 1.2 vs. 1.2 +/- 0.4; necrosis (foci/mm(2)), 3.9 +/- 2.3 vs. 0.4 +/- 0.3; inflammation (cells/mm(2)), 19.0 +/- 6.3 vs. 1.8 +/- 0.6. Centrilobular collagen deposition (% area), assessed by Sirius Red staining, was greater in Etoh vs. FD. Levels of endotoxin, ALT, CYP2E1 and lipid peroxidation markers were also higher (p < 0.01) in Etoh vs. FD. Levels of NF-kappaB and mRNA of pro-inflammatory mediators (TNF-alpha, COX-2, iNOS) and procollagen-I were increased (p < 0.05) in ethanol-fed rats. Immunohistochemical analysis showed more intense staining for both iNOS and nitrotyrosine in the centrilobular areas in the Etoh vs. FD groups. The greater area of positive staining for iNOS and nitrotyrosine in Etoh vs. FD was confirmed by Western Blot analysis. An increase in the expression of mRNA for profibrogenic genes (p < 0.05) was seen in ethanol-fed rats. A voluntary feeding regimen consisting of fish oil and ethanol in female rats is technically less demanding yet produces pathological and biochemical changes similar to those observed with the IG model. Pathological changes include fatty liver, necrosis and inflammation. Increased NF-kappaB and mRNA and protein levels of the pro-inflammatory mediators TNF-alpha, COX-2 and iNOS, coincided with the presence of necroinflammatory changes. The voluntary feeding regimen is proposed as an alternative to the IG model in the study of alcoholic liver injury.

Dietary fish oil down‐regulates pro‐inflammatory gene expression in colonocytes

Inflammatory bowel disease increases the risk of colon cancer development. The mechanisms behind this are based on the cellular consequences of oxidative stress. We have previously shown that dietary fish oil protects against inflammation-induced oxidative DNA damage by upregulation of apoptosis. To determine the biological processes ultimately responsible for the protective effect of dietary fish oil on inflammation, we determined the transcriptional profile in colonocytes using CodeLink microarrays. Sprague Dawley rats were provided with corn or fish oil containing diets for 3 wk. Dietary fish oil increased glutathione S-transferase (p= 0.007) expression, but decreased inflammation-related gene RelA (NFkB, p65) (p=0.0001), NFkB (p105) (p=0.006) and RNY 18366 (p=0.0007) compared to the corn oil group. Fish oil-fed rats had lower levels of CDP-diacylglycerol synthase expression (p=0.005), which is a key enzyme in the regulation of PIP2, and p21-activated kinase 2 and SH3 domain binding protein CR16, which are associated with MAP kinase signaling. The dietary fish oil group had greater bax expression (p=0.0009) compared to the corn oil group, which may contribute to the observed greater apoptosis in fish oil rats. In conclusion, dietary fish oil may protect against colon cancer by decreasing pro-inflammatory genes involved in the NFkB-, MAP kinase- and phosphoinositide-mediated signaling pathways. This may account for the previously observed lower oxidative DNA damage level in fish oil-fed rats and contribute towards one protective mechanism of fish oil against colon cancer. Funded by NIH (CA61750, CA82907, CA59034), NSBRI NASA NCC 9-58 and NIEHS P30-ES09106.

Boron deprivation alters rat behaviour and brain mineral composition differently when fish oil instead of safflower oil is the diet fat source*

Primary objective : To determine whether boron deprivation affects rat behaviour and whether behavioural responses to boron deprivation are modified by differing amounts of dietary long-chain omega-3 fatty acids. Research design : Female rats were fed diets containing 0.1 mg (9 μmol)/kg boron in a factorial arrangement with dietary variables of supplemental boron at 0 and 3 mg (278 μmol)/kg and fat sources of 75 g/kg safflower oil or 65 g/kg fish (menhaden) oil plus 10 g/kg linoleic acid. After 6 weeks, six females per treatment were bred. Dams and pups continued on their respective diets through gestation, lactation and after weaning. Between ages 6 and 20 weeks, behavioural tests were performed on 13–15 male offspring from three dams in each dietary treatment. The rats were euthanized at age 21 weeks for the collection of tissues and blood. Methods and procedures : At ages 6 and 19 weeks, auditory startle was evaluated with an acoustic startle system and avoidance behaviour was evaluated by using an elevated plus maze. At ages 7 and 20 weeks, spontaneous behaviour activity was evaluated with a photobeam activity system. A brightness discrimination test was performed on the rats between age 15 and 16 weeks. Brain mineral composition was determined by coupled argon plasma atomic emission spectroscopy. Plasma total glutathione was determined by HPLC and total cholesterol and 8- iso -prostaglandin F 2 (8- iso -PGF 2 ) were determined by using commercially available kits. Main outcomes and results : Boron-deficient rats were less active than boron-adequate rats when fed safflower oil based on reduced number, distance and time of horizontal movements, front entries, margin distance and vertical breaks and jumps in the spontaneous activity evaluation. Feeding fish oil instead of safflower oil attenuated the activity response to boron deprivation. In the plus maze evaluation, the behavioural reactivity of the boron-deficient rats fed fish oil was noticeably different than the other three treatments. They made more entries into both open and closed arms and the center area and thus visited more locations. The boron-deficient rats fed fish oil also exhibited the lowest copper and zinc and highest boron concentrations in brain and the highest plasma glutathione concentration. Both boron deprivation and safflower oil increased plasma 8- iso -PGF 2 . Conclusions : Both dietary boron and long-chain omega-3 fatty acids influence rat behaviour and brain composition and the influence of one these bioactive substances can be altered by changing the intake of the other. Brain mineral and plasma cholesterol, glutathione and 8- iso -PGF 2 findings suggest that rat behaviour is affected by an interaction between boron and fish oil because both affect oxidative metabolism and act the cellular membrane level.

Docosahexaneoic acid (22:6,n-3) regulates rat hepatocyte SREBP-1 nuclear abundance by Erk- and 26S proteasome-dependent pathways

Insulin induces and dietary n-3 PUFAs suppress hepatic de novo lipogenesis by controlling sterol-regulatory element binding protein-1 nuclear abundance (nSREBP-1). Our goal was to define the mechanisms involved in this regulatory process. Insulin treatment of rat primary hepatocytes rapidly augments nSREBP-1 and mRNA(SREBP-1c) while suppressing mRNA(Insig-2) but not mRNA(Insig-1). These events are preceded by rapid but transient increases in Akt and Erk phosphorylation. Removal of insulin from hepatocytes leads to a rapid decline in nSREBP-1 [half-time (T1/2) approximately 10 h] that is abrogated by inhibitors of 26S proteasomal degradation. 22:6,n-3, the major n-3 PUFA accumulating in livers of fish oil-fed rats, suppresses hepatocyte levels of nSREBP-1, mRNA(SREBP-1c), and mRNA(Insig-2) but modestly and transiently induces mRNA(Insig-1). More importantly, 22:6,n-3 accelerates the disappearance of hepatocyte nSREBP-1 (T1/2 approximately 4 h) through a 26S proteasome-dependent process. 22:6,n-3 has minimal effects on microsomal SREBP-1 and sterol-regulatory element binding protein cleavage-activating protein or nuclear SREBP-2. 22:6,n-3 transiently inhibits insulin-induced Akt phosphorylation but induces Erk phosphorylation. Inhibitors of Erk phosphorylation, but not overexpressed constitutively active Akt, rapidly attenuate 22:6,n-3 suppression of nSREBP-1. Thus, 22:6,n-3 suppresses hepatocyte nSREBP-1 through 26S proteasome- and Erk-dependent pathways. These studies reveal a novel mechanism for n-3 PUFA regulation of hepatocyte nSREBP-1 and lipid metabolism.

Influence of mitochondrial membrane fatty acid composition on proton leak and H 2O 2 production in liver

Mitochondrial membrane fatty acid composition has been proposed to play a role in determining mitochondrial proton leak rate. The purpose of this study was to determine if feeding rats diets with different fatty acid sources produces changes in liver proton leak and H 2O 2 production. Six-month-old male FBNF 1 rats were fed diets with a primary fat source of either corn or fish oil for a 6-month period. As expected, diet manipulations produced substantial differences in mitochondrial fatty acid composition. These changes were most striking for 20:4 n6 and 22:6 n3. However, proton leak and phosphorylation kinetics as well as lipid and protein oxidative damage were not different ( P>0.10) between fish and corn oil groups. Metabolic control analysis, however, did show that control of both substrate oxidation and phosphorylation was shifted away from substrate oxidation reactions to increased control by phosphorylation reactions in fish versus corn oil groups. Increased mitochondrial H 2O 2 production was observed in corn versus fish oil-fed rats when mitochondria were respiring on succinate alone or on either succinate or pyruvate/malate in the presence of antimycin A. These results show that mitochondrial H 2O 2 production and the regulation of oxidative phosphorylation are altered in liver mitochondria from rats consuming diets with either fish or corn oil as the primary lipid source.

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 approximately 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.