Cytochrome P450 epoxygenase-derived EPA and DHA oxylipins 17, 18-epoxyeicosatetraenoic acid and 19, 20-epoxydocosapentaenoic acid promote BAT thermogenesis and WAT browning through GPR120-AMPKα signaling pathway

The mechanisms whereby fish oil rich in EPA and DHA promotes BAT thermogenesis and WAT browning are not fully understood. Thus, this study aimed to investigate the effects of cytochrome...

Formation of Self-Assembled Liquid Crystalline Nanoparticles and Absorption Enhancement of Ω-3s by Phospholipids and Oleic Acids

This study aimed to optimize and evaluate self-assembled liquid crystalline nanoparticles (SALCs) prepared from phospholipids and oleic acid for enhancing the absorption of ω-3s. We explored the structure and optimal formulation of SALCs, which are composed of ω-3 ethyl ester (ω-3 EE), phospholipids, and oleic acid, using a ternary diagram and evaluated the improvement in ω-3 dissolution, permeation, and oral bioavailability. The in vitro dissolution and pharmacokinetics of ω-3 SALCs were compared with those of Omacor soft capsules (as the reference). The shape of the liquid crystal was determined according to the composition of phospholipids, oleic acids, and ω-3s and was found to be in cubic, lamellar, and hexagonal forms. The dissolution rates of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) obtained from ω-3 SALCs were 1.7 to 2.3-fold higher than those of the Omacor soft capsules. Furthermore, a pharmacokinetic study in male beagle dogs revealed that ω-3 SALCs increased the oral bioavailability of ω-3 EE by 2.5-fold for EPA and 3.1-fold for DHA compared with the reference. We found an optimal formulation that spontaneously forms liquid crystal-based nanoparticles, improving the bioavailability of EPA and DHA, not found in the existing literature. Our findings offer insight into the impact of nanoparticle phase on the oral delivery of oil-soluble drugs and provide a novel ω-3 EE formulation that improves the bioavailability of EPA and DHA.

Influence of fatty acids composition in different tissue of mice feeds with fish oils

Omega-3 long-chain polyunsaturated fatty acids (n-3 LC-PUFA), such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) play an important role in human health. Fish oils enriched with EPA and DHA have commercialized in triacylglycerol (TAG) and ethyl ester forms (EE). In this study, we compared the impact of diets containing fish oil in ethyl ester and triacylglycerol forms as a lipid source in five different tissues as liver, skeleteral muscle, brain, and epididymal white adipose tissue (WAT). The DHA levels were higher in the WAT and skeletal muscle of TAG and EE groups in comparison with the SB group. The body weight and brain, liver, epididymal WAT, and gastrocnemius muscle weights, and serum glucose, TG, cholesterol were not different between the groups. Thus, we conclude that EPA and DHA in the form of EE or TAG influence the fatty acids composition of different tissues.

Oral Bioavailability of Omega-3 Fatty Acids and Carotenoids from the Microalgae Phaeodactylum tricornutum in Healthy Young Adults

The microalgae Phaeodactylum tricornutum (PT) contains valuable nutrients such as proteins, polyunsaturated omega-3 fatty acids (n-3 PUFA), particularly eicosapentaenoic acid (EPA) and some docosahexaenoic acid (DHA), carotenoids such as fucoxanthin (FX), and beta-glucans, which may confer health benefits. In a randomized intervention trial involving 22 healthy individuals, we administered for two weeks in a crossover manner the whole biomass of PT (5.3 g/day), or fish oil (FO) containing equal amounts of EPA and DHA (together 300 mg/day). In an additional experiment, sea fish at 185 g/week resulting in a similar EPA and DHA intake was administered in nine individuals. We determined the bioavailability of fatty acids and carotenoids and assessed safety parameters. The intake of PT resulted in a similar increase in the n-3 PUFA and EPA content and a decrease in the PUFA n-6:n-3 ratio in plasma. PT intake caused an uptake of FX that is metabolized to fucoxanthinol (FXOH) and amarouciaxanthin A (AxA). No relevant adverse effects occurred following PT consumption. The study shows that PT is a safe and effective source of EPA and FX—and likely other nutrients—and therefore should be considered as a future sustainable food item.

Diverse Krill Lipid Fractions Differentially Reduce LPS-Induced Inflammatory Markers in RAW264.7 Macrophages In Vitro

Antarctic krill oil is an emerging marine lipid and expected to be a potential functional food due to its diverse nutrients, such as eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), phospholipids, astaxanthin and tocopherols. Although krill oil has been previously proved to have anti-inflammatory activity, there is little information about the relationship between its chemical compositions and anti-inflammatory activity. In this study, the RAW264.7 macrophages model was used to elucidate and compare the anti-inflammatory potential of different krill lipid fractions: KLF-A, KLF-H and KLF-E, which have increasing phospholipids, EPA and DHA contents but decreasing astaxanthin and tocopherols levels. Results showed that all the krill lipid fractions alleviated the inflammatory reaction by inhibition of production of nitric oxide (NO), release of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β and IL-6 and gene expression of proinflammatory mediators including TNF-α, IL-1β, IL-6, cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS). In addition, KLF-E with the highest phospholipids, EPA and DHA contents showed the strongest inhibition effect on the LPS-induced proinflammatory mediator release and their gene expressions. The results would be helpful to provide powerful insights into the underlying anti-inflammatory mechanism of krill lipid and guiding the production of krill oil products with tailor-made anti-inflammatory activity.

Method Development and Validation for Omega-3 Fatty Acids (DHA and EPA) in Fish Using Gas Chromatography with Flame Ionization Detection (GC-FID)

Gas chromatography with flame ionization detection (GC-FID) has often been used to quantify fatty acids in fish. This study validated the common method for determining omega-3 fatty acids (DHA and EPA) in the raw and cookedwarm-water fish, selayang, using GC-FID for subsequent evaluation on EPA and DHA retention using the Weibull model. The EPA and DHA were separated using a high-polarity capillary GC HP-88 column (60 m length, 0.25 mm ID, 0.2 μm DF) with a total run time of 45.87 min. The method was validated in linearity, precision, accuracy, specificity and sensitivity based on ICH requirements. In addition, it was found that the method had a high recovery rate (>95%) and good precision (RSD ≤ 2%) with overall RSDs ranging below 0.001% for both omega-3 PUFA. In conclusion, this method identified and quantified fatty acids and omega-3 accurately and precisely and can be used effectively for routine FAME analysis in fish samples.

Omega−3 Polyunsaturated Fatty Acids (PUFAs): Emerging Plant and Microbial Sources, Oxidative Stability, Bioavailability, and Health Benefits—A Review

The omega−3 (n−3) polyunsaturated fatty acids (PUFAs) eicosapentaenoic acid (EPA) and docosahexaenoic (DHA) acid are well known to protect against numerous metabolic disorders. In view of the alarming increase in the incidence of chronic diseases, consumer interest and demand are rapidly increasing for natural dietary sources of n−3 PUFAs. Among the plant sources, seed oils from chia (Salvia hispanica), flax (Linum usitatissimum), and garden cress (Lepidium sativum) are now widely considered to increase α-linolenic acid (ALA) in the diet. Moreover, seed oil of Echium plantagineum, Buglossoides arvensis, and Ribes sp. are widely explored as a source of stearidonic acid (SDA), a more effective source than is ALA for increasing the EPA and DHA status in the body. Further, the oil from microalgae and thraustochytrids can also directly supply EPA and DHA. Thus, these microbial sources are currently used for the commercial production of vegan EPA and DHA. Considering the nutritional and commercial importance of n−3 PUFAs, this review critically discusses the nutritional aspects of commercially exploited sources of n−3 PUFAs from plants, microalgae, macroalgae, and thraustochytrids. Moreover, we discuss issues related to oxidative stability and bioavailability of n−3 PUFAs and future prospects in these areas.

Changes in EPA and DHA during supplementation with omega-3 fatty acids and incident cardiovascular events: secondary analysis from the OMEMI trial

Background In the OMEMI trial, elderly post-MI patients did not achieve reduction in cardiovascular events from supplementation of 1.8g n-3 polyunsaturated fatty acids (PUFA). In two recent trials of hypertriglyceridaemic patients the REDUCE-IT trial demonstrated an association between high levels of serum eicosapentaenoic acid (EPA) and reduced risk of CV events with 4 g/day icosapent ethyl supplements while in the STRENGTH trial no such association was present in patients treated with 4 g/day of EPA+ docosahexaenoic acid (DHA). Purpose To assess associations between changes in concentrations of EPA and DHA during two years supplementation with n-3 PUFA and incident cardiovascular events in the OMEMI trial. Methods In the randomized controlled OMEMI trial, 1014 elderly patients with a recent acute myocardial infarction (AMI) were treated with 1.8g/day of EPA and docosahexaenoic acid (DHA) or placebo for two years, and followed for the primary outcome of MACE (AMI, coronary revascularization, stroke or heart failure hospitalization) and secondary outcome of new-onset atrial fibrillation (AF). Serum concentrations of EPA and DHA were measured at inclusion and at study completion by gas chromatography, and reported as % weight of total FA (%wt) in serum phospholipids. Results Serial EPA and DHA measurements at study inclusion and completion were available in 881 patients (92% of survivors). At baseline EPA and DHA concentrations were (mean±SD) 2.84±1.41 and 5.71±1.35%wt, respectively. Higher baseline EPA and DHA concentrations were associated with previous n-3 PUFA supplementation, lower prevalence of current smoking and diabetes, lower levels of triglycerides and higher levels of HDL-cholesterol (all p<0.05). In patients randomized to n-3 PUFA, EPA and DHA increased with 2.32±1.92 and 0.91±1.19%wt, respectively, whereas in the placebo group EPA and DHA decreased with −0.39±1.37 and −0.43±1.13%wt, respectively. Greater increases in EPA and DHA during follow-up were associated with a lowering of triglyceride concentrations, increasing HDL concentrations, and lower baseline concentrations of EPA and DHA (all p<0.001). Among patients treated with n-3 PUFA (n=438), a greater increase in EPA was associated with a lower risk of incident MACE (HR 0.89 [95% CI 0.78–1.00] per %wt, p=0.059) and higher risk of new-onset AF in patients free of AF at inclusion (n=339): HR 1.31 [1.06–1.62] per %wt, p=0.012 (Figure). There were no such associations for changes in DHA: HR 0.86 (95% CI 0.70–1.05), p=0.13 for MACE and HR 1.29 (0.91–1.83), p=0.16 for new-onset AF. Conclusion Patients treated with 1.8 g/day n-3 PUFA for two years experienced a doubling of serum EPA concentrations. Greater increases in EPA were associated with a lower risk of MACE, but also a higher risk of new-onset AF. Changes in DHA concentrations were not associated with outcomes, suggesting that EPA may be the more important n-3 PUFA with respect to risk of cardiovascular events. FUNDunding Acknowledgement Type of funding sources: Public hospital(s). Main funding source(s): Oslo University Hospital, Ullevål Figure 1

Omega-3 fatty acids differentially reduced expression of neutrophil degranulation-associated proteins in endothelial cells during IL-6 exposure

Background Neutrophil degranulation contributes to atherogenesis and tissue injury. Mixed omega-3 fatty acid (n3-FA) formulations of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have failed to reduce CV events compared to EPA only (REDUCE-IT), but the mechanisms are not understood. Purpose The purpose of this study was to compare the effects of EPA and DHA on expression of proteins linked to neutrophil degranulation in the vascular endothelium in vitro. Methods Human umbilical vein endothelial cells (HUVECs) were pretreated with EPA or DHA at equimolar levels (10 μM) for 2 h, then challenged with IL-6 at 12 ng/ml for 24 h. Proteomic analysis was performed using LC/MS to measure relative protein expression. Only significant (p<0.05) changes between treatment groups >1-fold were analyzed. Results In the Reactome “neutrophil degranulation” pathway, EPA and DHA downregulated 27 and 14 proteins, respectively, (p=9.97E-9 and 5.30E-4, respectively) relative to IL-6 alone. There were 12 protein changes common to both n3-FAs, including heme oxygenase-2 and ferritin light chain. EPA downregulated 15 proteins unlike DHA, including peroxiredoxin-6 and mitogen-activated protein kinase-1 (MAPK1). A combined 21 proteins downregulated by EPA and DHA versus IL-6 were upregulated by IL-6 alone relative to vehicle. EPA also increased expression of Rho-associated protein kinase-1 (ROCK-1), a protein downregulated by IL-6 alone and unaffected by DHA. Conclusions EPA and DHA differentially modulated expression of proteins linked to neutrophil degranulation. The distinct effects of EPA on protein expression may contribute to reduced inflammation in vascular injury compared to DHA. FUNDunding Acknowledgement Type of funding sources: Private company. Main funding source(s): Amarin Pharma Inc., Elucida Research