Feeding rats with liposomes or fish oil differently affects their lipid metabolism

Two n‐3 polyunsaturated fatty acid (PUFA)‐rich diets differing in their chemical and physical forms were given to rats during 2 wk. Liposomes [phospholipids (PL) organized in bilayer structures] made from a natural marine lipid extract or a mixture of fats containing fish oil [similar fatty acids esterified in triacylglycerols (TAG)] were used. The influence of n‐3 PUFA dietary sources on plasma parameters, i.e. TAG, cholesterol and PL concentrations, was investigated. A similar hypotriglyceridemic effect of n‐3 PUFA from liposomes or fish oil was observed. In contrast, feeding rats with liposomes led to different PL and cholesterol patterns. In the plasma of rats fed liposomes, total cholesterol amounts were positively correlated with PL levels. Liposome and fish oil feedings caused a marked increase in the amounts of n‐3 PUFA, which occurred mainly at the expense of n‐6 PUFA. However, the enrichment in n‐3 PUFA in the different plasma lipid classes differed substantially when ingested in the form of fish oil or liposomes. These results were interpreted in terms of different lipid bioavailability and metabolic fate during the digestive steps and in the liver, with the dietary source.     

Marine lipid-based liposomes increase <Emphasis Type="Italic">in vivo</Emphasis> FA bioavailability

Liposomes made from an extract of natural marine lipids and containing a high n-3 PUFA lipid ratio were envisaged as oral route vectors for FA supplements in order to increase PUFA bioavailability. The absorption of FA in thoracic lymph duct-cannulated rats, after intragastric feeding of dietary fats in the form of liposomes or fish oil, was compared. Lipid and FA analyses were also performed on feces. Five mole percent α-tocopherol was added to fish oil and incorporated into the liposome membrane. The influence of α-tocopherol on FA lymph recovery was also investigated. In vivo, FA absorption in rats was favored by liposomes (98±1%) compared to fish oil (73±6%). In the same way, the DHA proportion in lymph was higher after liposome ingestion (78%) than after fish oil ingestion (47%). However, phospholipid (PL) concentration in lymph was not affected by the kind of dietary fat ingested, suggesting a PL regulation due to de novo TAG synthesis. The influence of the intramolecular distribution of n-3 PUFA in dietary lipids (TAG and PL) on the intramolecular FA distribution in TAG of chylomicrons was also investigated. The results obtained showed that the distribution of n-3 PUFA esterified on the sn-2 of chylomicron TAG depended on the lipid source administered. All these results correlated, at least partly, with in vitro liposome behavior under conditions that mimic those of the gastrointestinal tract. As a whole, this study pointed out that marine PL may constitute an attractive material for the development of liposomes as oral PUFA supplements.     

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.     

Early Dissimilar Fates of Liver Eicosapentaenoic Acid in Rats Fed Liposomes or Fish Oil and Gene Expression Related to Lipid Metabolism

The study was undertaken to determine whether eicosapentaenoic acid (EPA, 20:5 n-3) and docosahexaenoic acid (DHA, 22:6 n-3), esterified in phospholipids (PL) as liposomes or in triglycerides (TG) as oil, exhibited comparable fates in liver lipids and whether these fates were associated with gene expressions related to fatty acid (FA) metabolism. PL and TG mixtures with close contents in EPA and DHA were administered to rats over 2 weeks. Most relevant events occurred after 3 days for both treatments. At that time, liposomes, compared with oil, increased the liver content in PL with a FA composition enriched in n-6 FA, comparable in DHA and much lower in EPA. Moreover, liposomes increased the activity and mRNA levels of carnitine palmitoyltransferase (CPT) I. In contrast, fish oil exerted opposite effects on CPT I and increased the genic expression of lipogenic enzymes. Liposomes, unlike fish oil, apparently increased the mRNA levels of acyl-CoA oxidase and the activity of the peroxisomal FA-oxidising system. Concomitantly, mRNA levels of hepatic lipoprotein receptors were increased with both diets, but intracellular proteins involved in free FA uptake and lipid synthesis were up-regulated only with liposome-treated rats. The quasi absence of EPA in hepatic PL of liposome-treated rats on the short term could result from increased β-oxidation activities through metabolic regulations induced by more available free EPA and other PUFA.     

Hypolipidemic Effects of Phospholipids (PL) Containing n-3 Polyunsaturated Fatty Acids (PUFA) Are Not Dependent on Esterification of n-3 PUFA to PL

Phospholipids (PL) containing n‐3 polyunsaturated fatty acids (PUFA) have beneficial effects of maintaining and promoting health compared with triacylglycerols (TAG) containing n‐3 PUFA or general PL. This study evaluated the effects of dietary PL containing n‐3 PUFA and elucidated the effects of the glycerophosphate structure and n‐3 PUFA on fatty acid (FA) metabolism in rats. Rats were fed a basal diet containing soybean oil alone, TAG containing n‐3 PUFA (1.8 %), soybean PL (2.7 %), PL containing n‐3 PUFA (2.7 %), or TAG containing n‐3 PUFA (1.8 %) + soybean PL (2.7 %). The present n‐3 PUFA‐supplemented diets had similar FA compositions, and the PL diets had similar PL compositions. TAG containing n‐3 PUFA reduced serum TAG contents, but did not affect serum cholesterol contents compared with soybean oil alone. PL diets containing n‐3 PUFA and the combination of TAG containing n‐3 PUFA and soybean PL resulted in decreased serum and liver TAG contents compared with the diet containing soybean oil alone, reflecting enhanced liver FA β‐oxidation. The results of this study show that TAG containing n‐3 PUFA with added soybean PL affects serum and liver TAG and cholesterol contents to a similar degree as PL containing n‐3 PUFA. TAG containing n‐3 PUFA and soybean PL are widely used as functional food ingredients and pharmaceutical constituents and are inexpensive compared with PL containing n‐3 PUFA. Therefore, the combination of TAG containing n‐3 PUFA and soybean PL has potential as a useful and inexpensive component of functional foods.     

Lymphatic fatty acids from rats fed human milk and formula supplemented with fish oil

Absorption of long-chain polyunsaturated fatty acids from human milk and formula supplemented with fish oil was studied to determine if the distribution route into lymphatic triacylglycerol (TAG) and phospholipid (PL) varies with the dietary source. Rats were intraduodenally infused with human milk or formula containing graded amounts of fish oil (0, 0.5, or 1.0 g/100 mL), and the mesenteric lymph was collected. Arachidonic acid (20∶4n−6) levels in lymphatic TAG and PL were highest from animals fed human milk. In the animals infused with formula containing fish oil, as the amount of eicosapentaenoic acid (EPA, 20∶5n−3) infused increased, there was essentially an equal increase of EPA associated with both lymphatic TAG and PL. Animals intraduodenally infused with human milk or formula without fish oil had only minor levels (less than 1%) of EPA in the lymph. In the fish oil-treated animals, as the amount of docosahexaenoic acid (DHA, 22∶6n−3) infused increased, there was a 16-fold increase in DHA associated with lymphatic TAG, but only a 3-fold increase in DHA associated with lymphatic PL. The highest level of DHA in rats infused with human milk was observed in lymphatic PL. Hence, fish oil can be added to formula as a source of long-chain polyunsaturated fatty acids, but the distribution of fatty acids into lymphatic TAG and PL is not the same as that observed with human milk.     

Analysis of EPA and DHA and distinction between fish oils and concentrates

Fish oil is the major dietary source of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Adequate dietary intake of these n‐3 PUFA is beneficial to reduce the risks of cardiovascular mortality, prostate cancer, and neurological disorders in children and adults. There is a surge in demand for fish oil in the functional food market. Microencapsulation of fish oil is the trend to improve its stability and sensory quality. The EPA and DHA content of the fish oil products may vary markedly from the label due to their susceptibility to oxidation. Quick and reliable methods other than the AOAC BF₃ method have been exploited to quantify EPA and DHA in the encapsulated fish oil and the microencapsulated powdered products such as infant formula. This article describes a method to differentiate the ethyl‐ester form from the TAG form of EPA and DHA in encapsulated fish oil which may be a mixture of natural triacylglycerol enriched with ethyl esterified EPA and DHA. A method is recommended due to the difference in apparent potency of these two esterified forms, which may be a concern in infant formula and elsewhere.     

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.     

n‐3 PUFA Induce Microvascular Protective Changes During Ischemia/Reperfusion

Ischemia/reperfusion (I/R) injury can occur in consequence of myocardial infarction, stroke and multiple organ failure, the most prevalent cause of death in critically ill patients. I/R injury encompass impairment of endothelial dependent relaxation, increase in macromolecular permeability and leukocyte‐endothelium interactions. Polyunsaturated fatty acids (n‐3 PUFA), such as eicosapentaenoic acid (EPA, 20:5n‐3) and docosahexaenoic acid (DHA, 22:6n‐3) found in fish oil have several anti‐inflammatory properties and their potential benefits against I/R injury were investigated using the hamster cheek pouch preparation before and after ischemia. Before the experiments, hamsters were treated orally with saline, olive oil, fish oil and triacylglycerol (TAG) and ethyl ester (EE) forms of EPA and DHA at different daily doses for 14 days. Fish oil restored the arteriolar diameter to pre ischemic values during reperfusion. At onset and during reperfusion, Fish oil and DHA TAG significantly reduced the number of rolling leukocytes compared to saline and olive oil treatments. Fish oil, EPA TAG and DHA TAG significantly prevented the rise on leukocyte adhesion compared to saline. Fish oil (44.83 ± 3.02 leaks/cm²), EPA TAG (31.67 ± 2.65 leaks/cm²), DHA TAG (41.14 ± 3.63 leaks/cm²), and EPA EE (30.63 ± 2.25 leaks/cm²), but not DHA EE (73.17 ± 2.82 leaks/cm²) prevented the increase in macromolecular permeability compared to saline and olive oil (134.80 ± 1.49 and 121.00 ± 4.93 leaks/cm², respectively). On the basis of our findings, we may conclude that consumption of n‐3 polyunsaturated fatty acids, especially in the triacylglycerol form, could be a promising therapy to prevent microvascular damage induced by ischemia/reperfusion and its consequent clinical sequelae.     

Uncoupling EPA and DHA in Fish Nutrition: Dietary Demand is Limited in Atlantic Salmon and Effectively Met by DHA Alone

Due to the scarcity of marine fish oil resources, the aquaculture industry is developing more efficient strategies for the utilization of dietary omega‐3 long‐chain polyunsaturated fatty acids (n‐3 LC‐PUFA). A better understanding of how fish utilize EPA and DHA, typically provided by fish oil, is needed. However, EPA and DHA have different physiological functions, may be metabolized and incorporated into tissues differently, and may vary in terms of their importance in meeting the fatty acid requirements of fish. To address these questions, Atlantic salmon were fed experimental diets containing, as the sole added dietary lipid source, fish oil (positive control), tallow (negative control), or tallow supplemented with EPA, DHA, or both fatty acids to ~50 or 100 % of their respective levels in the positive control diet. Following 14 weeks of feeding, the negative control diet yielded optimum growth performance. Though surprising, these results support the notion that Atlantic salmon requirements for n‐3 LC‐PUFA are quite low. EPA was largely β‐oxidized and inefficiently deposited in tissues, and increasing dietary levels were associated with potential negative effects on growth. Conversely, DHA was completely spared from catabolism and very efficiently deposited into flesh. EPA bioconversion to DHA was largely influenced by substrate availability, with the presence of preformed DHA having little inhibitory effect. These results clearly indicate EPA and DHA are metabolized differently by Atlantic salmon, and suggest that the n‐3 LC‐PUFA dietary requirements of Atlantic salmon may be lower than reported and different, if originating primarily from EPA or DHA.     

Replacement of Marine Fish Oil with de novo Omega-3 Oils from Transgenic Camelina sativa in Feeds for Gilthead Sea Bream (Sparus aurata L.)

Omega‐3 (n‐3) long‐chain polyunsaturated fatty acids (LC‐PUFA) are essential components of the diet of all vertebrates. The major dietary source of n‐3 LC‐PUFA for humans has been fish and seafood but, paradoxically, farmed fish are also reliant on marine fisheries for fish meal and fish oil (FO), traditionally major ingredients of aquafeeds. Currently, the only sustainable alternatives to FO are vegetable oils, which are rich in C₁₈ PUFA, but devoid of the eicosapentaenoic (EPA) and docosahexaenoic acids (DHA) abundant in FO. Two new n‐3 LC‐PUFA sources obtained from genetically modified (GM) Camelina sativa containing either EPA alone (ECO) or EPA and DHA (DCO) were compared to FO and wild‐type camelina oil (WCO) in juvenile sea bream. Neither ECO nor DCO had any detrimental effects on fish performance, although final weight of ECO‐fed fish (117 g) was slightly lower than that of FO‐ and DCO‐fed fish (130 and 127 g, respectively). Inclusion of the GM‐derived oils enhanced the n‐3 LC‐PUFA content in fish tissues compared to WCO, although limited biosynthesis was observed indicating accumulation of dietary fatty acids. The expression of genes involved in several lipid metabolic processes, as well as fish health and immune response, in both liver and anterior intestine were altered in fish fed the GM‐derived oils. This showed a similar pattern to that observed in WCO‐fed fish reflecting the hybrid fatty acid profile of the new oils. Overall the data indicated that the GM‐derived oils could be suitable alternatives to dietary FO in sea bream.     

Effects of dietary triacylglycerol structure on triacylglycerols of resultant chylomicrons from fish oil- and seal oil-fed rats

We investigated the influence of the intramolecular fatty acid distribution of dietary triacyl-sn-glycerols (TAG) rich in n-3 polyunsaturated fatty acids (PUFA) on the structure of chylomicron TAG. Fish oil and seal oil, comparable in fatty acid compositions but with different contents of major n-3 PUFA esterified at thesn-2 position (20:5n-3, 46.6%, and 5.3%; 22:6n-3, 75.5%, and 3.8%, respectively), were fed to rats. Mesenteric lymph was collected and the chylomicrons were isolated by ultracentrifugation. The fatty acid composition of chylomicrons largely reflected the fatty acid composition of the oils administered. The intramolecular fatty acid distributions of the TAG fed were reflected in the chylomicron TAG as the fraction of the total contents observed in thesn-2 position of 20:5n-3 were 23.6 and 13.3%, and of 22:6n-3 were 30.6 and 5.4% for resultant chylomicrons following fish oil and seal oil administration, respectively. Thus, after seal oil administration, significant higher load of n-3 PUFA was esterified in thesn-1,3 positions of chylomicron TAG compared with fish oil administration (P<0.05).     

Lipase selectivity toward fatty acids commonly found in fish oil

The main objective of this study was to compare the fatty acid selectivity of numerous commercially available lipases toward the most ubiquitous fatty acids present in fish oils in form of their corresponding ethyl esters. Special interest was taken in their ability to separate the n‐3 long‐chain polyunsaturated fatty acids (PUFA), mainly eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), from the more saturated fatty acids as well as exploiting the putative discrimination between these highly valuable n‐3 PUFA. Hydrolysis of sardine oil ethyl esters in a Tris buffer solution by 12 microbial lipases is described. The results reveal that all of the lipases strongly discriminate against the n‐3 PUFA and prefer the more saturated fatty acids as substrates. Most of the lipases discriminate between EPA and DHA in favor of EPA, however, 2 bacterial lipases from Pseudomonas were observed to prefer DHA to EPA. Digestive lipolytic enzymes isolated from salmon and rainbow trout intestines displayed reversed fatty acid selectivity when their fish oil triacylglycerol hydrolysis was studied. Thus, the n‐3 PUFA including EPA and DHA were observed to be hydrolyzed at a considerably higher rate than the more saturated fatty acids.     

Dietary menhaden, seal, and corn oils differentially affect lipid and Ex vivo eicosanoid and thiobarbituric acid-reactive substances generation in the guinea pig

This investigation was carried out to characterize the effects of specific dietary marine oils on tissue and plasma fatty acids and their capacity to generate metabolites (prostanoids, lipid peroxides). Young male guinea pigs were fed nonpurified diet (NP), or NP supplemented (10%, w/w) with menhaden fish oil (MO), harp seal oil (SLO), or corn oil (CO, control diet) for 23 to 28 d. Only the plasma showed significant n−3 polyunsaturated fatty acid (PUFA)-induced reductions in triacylglycerol (TAG) or total cholesterol concentration. Proportions of total n−3 PUFA in organs and plasma were elevated significantly in both MO and SLO dietary groups (relative to CO), and in all TAG fractions levels were significantly higher in MO-than SLO-fed animals. The two marine oil groups differed in their patterns of incorporation of eicosapentaenoic acid (EPA). In guinea pigs fed MO, the highest levels of EPA were in the plasma TAG, whereas in SLO-fed animals, maximal incorporation of EPA was in the heart polar lipids (PL). In both marine oil groups, the greatest increases in both docosahexaenoic acid (22∶6n−3, DHA) and docosapentaenoic acid (22∶5n−3, DPA) relative to the CO group, were in plasma TAG, although the highest proportions of DHA and DPA were in liver PL and heart TAG, respectively. In comparing the MO and SLO groups, the greatest difference in levels of DHA was in heart TAG (MO>SLO, P<0.005), and in levels of DPA was in heart PL (SLO>MO, P<0.0001). The only significant reduction in proportions of the major n−6 PUFA, arachidonic acid (AA), was in the heart PL of the SLO group (SLO>MO=CO, P<0.005). Marine oil feeding altered ex vivo generation of several prostanoid metabolites of AA, significantly decreasing thromboxane A₂ synthesis in homogenates of hearts and livers of guinea pigs fed MO and SLO, respectively (P<0.04 for both, relative to CO). Lipid peroxides were elevated to similar levels in MO- and SLO-fed animals in plasma, liver, and adipose tissue, but not in heart preparations. This study has shown that guinea pigs respond to dietary marine oils with increased organ and plasma n−3 PUFA, and changes in potential synthesis of metabolites. They also appear to respond to n−3 PUFA-enriched diets in a manner that is different from that of rats.     

Dietary ALA from Spinach Enhances Liver n-3 Fatty Acid Content to Greater Extent than Linseed Oil in Mice Fed Equivalent Amounts of ALA

Although several works have reported absorption rate differences of n‐3 polyunsaturated fatty acids (PUFA) bound to different lipid forms, such as ethyl ester, triacylglycerol (TAG), and phospholipids, no studies have investigated the effect of n‐3 PUFA from glycolipids (GL). The present study compared the fatty acid contents of tissue and serum lipids from normal C57BL/6J mice fed two types of α‐linolenic acid (ALA)‐rich lipids, spinach lipid (SPL), and linseed oil (LO). ALA was primarily present as the GL form in SPL, while it existed as TAG in LO. Supplementation of both lipids increased ALA and its n‐3 metabolites, eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA), and docosahexaenoic acid, and decreased n‐6 PUFA, linoleic acid and arachidonic acid, in the livers, small intestines, and sera of the treated mice compared with those of the control group. When the comparison between the SPL and LO diets containing the same amount of ALA was conducted, the EPA and DPA levels in the liver lipids from mice fed the SPL diet were significantly higher than those fed the LO diet. Additionally, the total contents of n‐3 PUFA of lipids from the livers, small intestines, and sera of the SPL group were higher than those of the LO group.     

Lipid Profile and Glycerophospholipid Molecular Species in Two Species of Edible Razor Clams Sinonovacula constricta and Solen gouldi

Total lipids were extracted from razor clams Sinonovacula constricta and Solen gouldi, and the molecular species of glycerophospholipid (Gpl) including choline glycerophospholipid (ChoGpl), ethanolamine glycerophospholipid (EtnGpl), serine glycerophospholipid (SerGpl), inositol glycerophospholipid (InsGpl), lysoChoGpl, lysoEtnGpl, and lysoSerGpl were characterized using a direct-infusion tandem mass spectrometric method for the first time. Meanwhile, the lipid class composition and phospholipid (PL) class composition as well as the fatty acid (FA) composition of total lipids, triacylglycerol (TAG), and PL were also investigated. About 238 and 235 molecular species were characterized, respectively, in Sinonovacula constricta and Solen gouldi. The majority of the dominant Gpl molecular species contained n-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFA), especially docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA). Also, razor clam lipids contained a high-proportioned PL (52.19–65.41% of total lipids) and PUFA (47.94–54.81 mol%). Furthermore, PL contained a higher proportion of PUFA (63.05–67.13 mol%), especially DHA (20.04–22.47 mol%) and EPA (16.27–21.46 mol%) than TAG (the corresponding values being 33.73–34.45, 11.95–12.27, and 8.13–0.8.99 mol%, respectively). Meanwhile, phosphatidylcholine (44.38–46.21 mol%) and phosphatidylethanolamine (38.84–39.95 mol%) were dominant among PL. In consideration of the high proportion of PUFA-enriched Gpl, razor clam plays a great role in promoting human health.     

Specific modifications of phosphatidylinositol and nonesterified fatty acid fractions in cultured porcine cardiomyocytes supplemented with n-3 polyunsaturated fatty acids

Mechanisms for the antiarrhythmic effect of n−3 polyunsaturated fatty acids (PUFA) are currently being investigated using isolated cardiac myocytes. It is still not known whether the incorporation of n−3 PUFA into membrane phospholipids is a prerequisite for its protective action or if n−3 PUFA exert antiarrhythmic effects in their nonesterified form as demonstrated by recent studies. Adult porcine cardiomyocytes were grown in media supplemented with arachidonic acid (AA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA). After 24 h, analysis of total lipids showed that the myocytes were enriched with the respective fatty acids compared to control cells. Large proportions of all three fatty acids supplemented (69% AA, 72% DHA, and 66% EPA) remained unesterified. Fatty acid analysis of total phospholipids (PL) revealed that the incorporation of EPA and DHA, though small, was significantly different (P<0.05) from that of the control cells. The PL fraction was further separated into phosphatidylinositol (Pl), phosphatidylethanolamine, phosphatidylcholine, and phosphatidylserine to study the pattern of incorporation of the fatty acids in these fractions. It became apparent that EPA and DHA were selectively incorporated into the Pl fraction. This study demonstrates that in adult porcine cardiomyocytes, the n−3 PUFA supplementation selectively modulates two important lipid fractions, nonesterified fatty acid and Pl, which were implicated in the mechanisms of prevention of cardiac arrhythmias.     

EPA or DHA Supplementation Increases Triacylglycerol,but not Phospholipid,Levels in Isolated Rat Cardiomyocytes

It is well recognized that a high dietary intake of long-chain polyunsaturated fatty acids (LC-PUFA) has profound benefits on health and prevention of chronic diseases. In particular, in recent years there has been a dramatic surge of interest in the health effects of n-3 LC-PUFA derived from fish, eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids. Notwithstanding, the metabolic fate and the effects of these fatty acids once inside the cell has seldom been comprehensively investigated. Using cultured neonatal rat cardiomyocytes as model system we have investigated for the first time, by means of high-resolution magic-angle spinning nuclear magnetic resonance (HR-MAS NMR) spectroscopy in combination with gas chromatography (GC), the modification occurring in the cell lipid environment after EPA and DHA supplementation. The most important difference between control and n-3 LC-PUFA-supplemented cardiomyocytes highlighted by HR-MAS NMR spectroscopy is the increase of signals from mobile lipids, identified as triacylglycerols (TAG). The observed increase of mobile TAG is a metabolic response to n-3 LC-PUFA supplementation, which leads to an increased lipid storage. The sequestration of mobile lipids in lipid bodies provides a deposit of stored energy that can be accessed in a regulated fashion according to metabolic need. Interestingly, while n-3 LC-PUFA supplementation to neonatal rat cardiomyocytes causes a huge variation in the cell lipid environment, it does not induce detectable modifications in water-soluble metabolites, suggesting negligible interference with normal metabolic processes.     

Fatty Acid and Lipid Profiles with Emphasis on n-3 Fatty Acids and Phospholipids from Ciona intestinalis

In order to establish Ciona intestinalis as a new bioresource for n‐3 fatty acids‐rich marine lipids, the animal was fractionated into tunic and inner body tissues prior to lipid extraction. The lipids obtained were further classified into neutral lipids (NL), glycolipids (GL) and phospholipids (PL) followed by qualitative and quantitative analysis using GC‐FID, GC–MS, ¹H NMR, 2D NMR, MALDI‐TOF‐MS and LC–ESI–MS methods. It was found that the tunic and inner body tissues contained 3.42–4.08 % and 15.9–23.4 % of lipids respectively. PL was the dominant lipid class (42–60 %) irrespective of the anatomic fractions. From all lipid fractions and classes, the major fatty acids were 16:0, 18:1n‐9, C20:1n‐9, C20:5n‐3 (EPA) and C22:6n‐3 (DHA). The highest amounts of long chain n‐3 fatty acids, mainly EPA and DHA, were located in PL from both body fractions. Cholestanol and cholesterol were the dominant sterols together with noticeable amounts of stellasterol, 22 (Z)‐dehydrocholesterol and lathosterol. Several other identified and two yet unidentified sterols were observed for the first time from C. intestinalis. Different molecular species of phosphatidylcholine (34 species), sphingomyelin (2 species), phosphatidylethanolamine (2 species), phosphatidylserine (10 species), phosphatidylglycerol (9 species), ceramide (38 species) and lysophospholipid (5 species) were identified, representing the most systematic PL profiling knowledge so far for the animal. It could be concluded that C. intestinalis lipids should be a good alternative for fish oil with high contents of n‐3 fatty acids. The lipids would be more bioavailable due to the presence of the fatty acids being mainly in the form of PL.     

Fatty Acid Composition of Tropical Fish Depends on Reservoir Trophic Status and Fish Feeding Habit

Eutrophication results in a deficiency of n‐3 LC‐PUFA (long‐chain polyunsaturated fatty acids) in aquatic food chains, affecting fish nutrition and physiology. The trophic transfer of FA (fatty acids) to fish species of different feeding habits was investigated in two reservoirs in southeast Brazil—the mesotrophic Ponte Nova Reservoir (PN) and the hypereutrophic Billings Reservoir (Bil). Total FA profile of stomach contents and adipose tissue, triacylglycerols (TAG), and phospholipids (PL) from liver and muscle of the omnivorous Astyanax fasciatus and the carnivorous Hoplias malabaricus were analyzed by gas chromatography. A prevalence of n‐6PUFA, as 18:2n‐6 (linoleic acid) and 20:4n‐6 (arachidonic acid, ARA) was observed in the stomach contents and in the tissues of A. fasciatus from the PN reservoir. In contrast, n‐3 LC‐PUFA, as 20:5n‐3 (eicosapentaenoic acid, EPA) was accumulated in fish tissues from Bil, resulting in higher n3/n6 and EPA/ARA ratios, compared to fish from PN. This differential FA accumulation was also observed for H. malabaricus, but differences were slightly minor, and no changes were observed in the EPA/ARA ratios between fish from both reservoirs. Regardless reservoir, FA profiles of TAG resembled that of their diet, whereas FA profiles of PL were more conservative and mainly comprised by LC‐PUFA. We conclude that reservoir trophic status affected the FA composition of food resources available to these fish species, resulting in differential allocation of n‐3 and n‐6 FA. As expected, FA profile of the investigated fish species also reflected their feeding habit and physiological demands.