EPA Treatment Research Articles

Fatty acid-sensing mechanisms in the hypothalamus of European sea bass (Dicentrarchus labrax): The potential role of monounsaturated and polyunsaturated fatty acids

This study aimed to evaluate the hypothalamus fatty acid (FA)-sensing mechanisms response to different FA in European sea bass. For that purpose, fish (body weight of 90 g) were intraperitoneally (IP) injected (time 0 h) with five long-chain unsaturated fatty acids, namely, docosahexaenoic acid (DHA; C22:5n3); eicosapentaenoic acid (EPA; C20:4n3); α-linolenic (ALA; C18:3n3); linoleic acid (LA; C18:2n6) and oleic acid (OA; C18:1n9) at a dose of 300 μg kg−1, or with 0.9% saline solution (control). Feed intake (FI) was recorded at 3, 6, and 24 h after the IP injection. One week later, fish were IP injected with the same FA, and the hypothalamus was collected 3 h after the IP injection for measurement of molecules related to FI regulation and FA-sensing mechanisms. Cumulative FI (g/kg/day) was not affected by treatments. However, compared to the control, FI increased with the OA treatment at 6 h after the IP injection. FI decreased with mealtime in the DHA and LA groups. Gene expression of orexigenic (npy/agrp) and anorexigenic (cart2/pomc1) neurons was not affected by the FA treatments. Attending the enzymes involved in the FA-sensing mechanisms activation, compared to the control carnitine palmitoyltransferase I (CPT1) and ATP citrate lyase (ACLY) activity were not affected by FA treatments. Contrarily the key enzymes of lipid metabolisms, malic enzyme and hydroxyacylCoA dehydrogenase was higher in fish that received the EPA and OA treatment, than fish treated to the control. Overall, the results of the present study indicate that gene expression of orexigenic and anorexigenic neurons was not affected at 3 h after IP injection with different FA. However, the activity of key enzymes of lipid metabolism was differently affected by circulating FA, indicating that FA-sensing mechanisms respond to different FA. Further studies are required involving different sampling times to further characterize the response of FA-sensing mechanisms to FA. These findings may be of relevance to the aquaculture industry in an era where alternative lipid sources are being increasingly used.

Abstract 13683: Eicosapentaenoic Acid (EPA) Decreases Endothelin Converting Enzyme-1 Expression and Improves Nitric Oxide Release in Pulmonary Endothelium Following Challenge With Air Pollutants

Background: Air pollution particulate matter (PM) contributes to global mortality and chronic conditions, especially cardiovascular (CV) disease. Exposure to PM and cytokines like IL-6 cause endothelial cell (EC) dysfunction, leading to nitric oxide synthase (eNOS) uncoupling. The omega-3 fatty acid EPA improves eNOS coupling and reduces vascular inflammation. REDUCE-IT showed that treatment with icosapent ethyl (IPE), a purified EPA ethyl ester, reduced composite CV events. We compared the effects of EPA on pulmonary EC (PEC) function and protein expression following challenge with PMs versus IL-6. Methods: PECs were treated with EPA (40 μM) or equivolume vehicle for 2 h and then challenged with IL-6 (12 ng/mL) or PM (50 μg/mL) collected from urban environments (“urban PM”) for up to 8 h. Proteomic analysis was performed using LC/MS to measure relative protein expression. Significant ( p <0.05) changes between treatment groups (>1-fold) were analyzed by bioinformatic packages. Biological pathways were analyzed by gene set enrichment analysis (GSEA). In parallel, cells were stimulated with calcium ionophore to elicit nitric oxide (NO) and peroxynitrite (ONOO – ) release as assayed by tandem porphyrinic nanosensors; the NO/ONOO – ratio indicates eNOS coupling efficiency. Results: Challenge with IL-6 and urban PM caused eNOS uncoupling as evidenced by a decreased NO/ONOO – ratio of 35% (1.43 ± 0.04 vs. 2.19 ± 0.02, p <0.001) and 50% (1.22 ± 0.07 vs. 2.42 ± 0.15 , p <0.001), respectively, compared to control. EPA increased the NO/ONOO – ratio relative to both IL-6 (39%; 1.99 ± 0.11, p <0.001) and urban PM (39%; 1.69 ± 0.14 , p <0.05). Proteomics revealed that EPA decreased expression of endothelin converting enzyme-1 (ECE1) relative to both IL-6 (1.2-fold, p = 0.017) and urban PM (1.1-fold, p = 0.045). GSEA showed significant modulations to 18 and 35 proteins within the neutrophil degranulation pathway (GO:0043312) with EPA treatment relative to IL-6 and urban PM ( p <0.0001 in both comparisons). Conclusion: EPA favorably reversed EC dysfunction and expression of inflammatory proteins and pathways following challenge with air pollution PMs similar to IL-6. These broad anti-inflammatory actions may contribute to reduced CV risk with EPA as observed in outcome trials.

The Effects of Maternal Intake of EPA and DHA Enriched Diet During Pregnancy and Lactation on Offspring's Muscle Development and Energy Homeostasis.

EPA and DHA are n-3 long-chain polyunsaturated fatty acids with a diversity of health benefits on offspring. The objective of this study was to test the in vivo effect of maternal ingestion of EPA and DHA on fetal and offspring muscle development and energy balance. Two groups of female C57BL/6 mice were fed EPA and DHA enriched diet (FA) and diet devoid of EPA and DHA (CON) respectively throughout the entire period of gestation and lactation. Embryos at E13 and offspring at age of D1 and D21 were selected for sample collection and processing. No change in birth number and body weight were observed between groups at D1 and D21. Transient increase in the expression levels of myogenesis regulating genes was detected at D1 (p < 0.05) in FA group. Most of the expression of muscle protein synthesis regulating genes were comparable (p > 0.05) between FA and CON groups at D1 and D21. The significant increase in MHC4, and IGF-1 was not linked to increased muscle mass. A persistent increase in ISR expression (p < 0.05) but not in GLUT-4 (p > 0.05) was detected in offspring. Up-regulation of adipogenesis regulating genes was accompanied by increasing intramuscular fat accumulation in the offspring of FA group. Considerable increase in transcripts of genes regulating lipid catabolism and thermogenesis in liver (p < 0.05) was noticed in FA group at D21; whereas, only the levels of carnitine palmitoyl transferase 1A (Cpt1α) and Enoyl-CoA Hydratase And 3-Hydroxyacyl CoA Dehydrogenase (Ehhadh) increased at D1. Similarly, genes regulating lipolysis were highly expressed at D21 in FA group. EPA and DHA treatment promoted BAT development and activity by increasing the expression of BAT signature genes (p < 0.05). Also, maternal intake of EPA and DHA enriched diet enhanced browning of sWAT. Taken together, maternal ingestion of EPA/DHA may be suggested as a therapeutic option to improve body composition and counteract childhood obesity- related metabolic disorders and confer lifelong positive metabolic impact on offspring.

Omega-3 fatty acids and blood-based biomarkers in Alzheimer’s disease and mild cognitive impairment: A randomized placebo-controlled trial

BackgroundIncreased serum levels of pro-inflammatory biomarkers are consistently associated with cognitive decline. The omega-3 unsaturated fatty acids (n-3 PUFAs) had been linked to slowing cognitive decline due to their potential anti-inflammatory effects. To our knowledge, the different regiments of pure DHA, pure EPA, and their combination on various associated symptoms of dementia, including a mild form of cognitive impairment (MCI) and Alzheimer’s disease (AD), have never been studied. MethodsThis multisite, randomized, double-blind, placebo-controlled trial was conducted at two veteran’s retirement centers and one medical center in central Taiwan between 2013 and 2015. 163 MCI or AD patients were randomly assigned to placebo (n = 40), docosahexaenoic acid (DHA, 0.7 g/day, n = 41), eicosapentaenoic acid (EPA, 1.6 g/day, n = 40), or EPA (0.8 g/day) + DHA (0.35 g/day) (n = 42) group for 24 months. The results were measured as the cognitive and functional abilities, biochemical, and inflammatory cytokines profiles. Chi-square tests, two-sample t-test, ANOVA, and linear mixedeffects models were conducted with p < 0.05. Results131 (80%) participants had completed the trial with all cognitive, functional, and mood status assessments. The statistically significant difference between the placebo and treatment groups was not determined, concerning the changes in cognitive, functional, and mood status scores, the biochemical profiles, and inflammatory cytokines levels. However, EPA was found to reduce the C–C motif ligands 4 (CCL4) level (p < 0.001). Additionally, EPA could reduce the constructional praxis (p < 0.05) and spoken language ability scores (p < 0.01), and DHA also reduced the spoken language ability score (p < 0.05). ConclusionOverall, n-3 PUFAs supplements did not reduce cognitive, functional, and depressive symptom outcomes, but spoken language ability and constructional praxis subitems of ADAS-cog. These findings show that attention to clinical heterogeneity in dementia is crucial when studying nutrients interventions, such as n-3 PUFAs. In addition, with small effect size CCL4 is a better indicator than other inflammatory cytokines for EPA treatment response.

Effect of EPA on Hsp90 and GR\u03b1 protein expression in multiple myeloma drug-resistant cells

BackgroundApproximately 20% of MM patients harbor glucocorticoid (GC) resistance and are not responsive to therapeutic effect. Chaperoneheat-shock proteins Hsp90 is needed for ligand docking, The imbalance of Hsp90/GRα (glucocorticoid receptor α) may be an important cause of GC resistance. Recent studies have indicated that EPA could repress cancer cell growth by regulating critical influential factors in progression of cancer, consisting of resistance to drugs, chemosensitivity. The aim of the present study was to test the cytotoxic effects of EPA alone or EPA + Dexamethasone in dexamethasone-resistant MM cell (MM.1R) and investigate whether DHA can induce apoptosis and reverse acquired glucocorticoid resistance in dexamethasone-resistant MM cell (MM.1R).MethodsCell Counting Kit-8 (CCK-8) was used to detect the proliferation of MM.1R cells after treating with EPA alone and EPA combined with DEX. Mitochondrial membrane potential was measured by flow cytometry and GRα and Hsp90 protein expression were assessed by western blot analysis.ResultsEPA alone was able to inhibit cell proliferation as evidenced by CCK-8 assay and the tumor growth was remarkably suppressed by EPA + Dexamethasone, Cell apoptosis after EPA treatment was obviously observed by Flow cytometry analysis of the mitochondrial membrane potential. Analysis of Hsp90 and GRα proteins in MM.1R cells incubated with EPA revealed down-regulation of Hsp90 and up-regulation of GRα. Accordingly, the Hsp90/GRα ratio was significantly decreased with the increase of EPA concentration.ConclusionsEPA might be used as a new effective treatment for reversal of glucocorticoid-resistance in multiple myeloma.

The Physiological and Metabolic Effects of Maternal Intake of EPA and DHA during Pregnancy and Lactation on Offspring's Muscle Development and Energy Homeostasis

EPA and DHA are long-chain fatty acids with health benefits on offspring underpinning the accumulative recommendations for their prenatal ingestion. Recent in vivo studies have emerged indicating a positive correlation of EPA and DHA with fetal BAT development, while other in vitro trials demonstrated their negative implication in promoting fetal myoblast reprogramming into adipoblasts. Therefore, the objective of this study was to test the effect of maternal EPA and DHA on fetal muscle development and energy balance. Two groups of Female C57BL/6 mice were assigned to different diets throughout the entire period of gestation and lactation; the first group was fed EPA and DHA enriched diet (FA); whereas, the other was fed a diet devoid of n-3 PUFA (CON). Embryos at day 13 of gestation and offspring at age of 1 and 21day were selected for sample collection and processing. No change in little number and body weight were observed between groups in newborns at day1, but a tendency to weigh loss in the FA group was detected after weaning. Transient increase in the expression levels of MYOD1, MYOG, MRF4, and MHC4 were observed at day 1 post-parturition (P<0.05). Except of IGF-1 P<0.05, the expression levels of genes regulating muscle protein synthesis and catabolism were comparable (P>0.05) between groups at day 1 and 21. Persistent increase in ISR expression (P<0.05) but not in GLUT-4 (P>0.05) were detected in day 1 and 21 age offspring. An increase in the expression of PPARγ and AP2 but not C/EBPα (P<0.05) at day 1 were observed. The three key regulators of basal adipogenesis were up-regulated in weaned mice (P<0.05). Up-regulation of PPARγ, AP2, and C/EBPα levels were accompanied by increasing intramuscular fat accumulation in day 1 and 21 samples. Considerable increase in the transcripts of genes regulating lipid catabolism and thermogenesis in liver including Cpt1α,Ehhadh, Mcad, Lcad, Acadvl, slc22a5, slc25a20, and Pparα (P<0.05) was noticed in (FA) group after 21 day of treatment; whereas, only the transcripts of Cpt1α andEhhadh increased in day1 neonates’ livers . Similarly, genes regulating lipolysis (ATGL and HSL, MGL and LPL) were up-regulated at day 21 of treatment only in (FO) group (P<0.05) while EPA and DHA maternal intake does not affect the expression of lipogenesis regulating genes (Fasn and Srebp1c) (P>0.05) in day 1 and 21 offspring. EPA and DHA treatment promoted BAT development and activity through increasing the expression of BAT signature genes including (UCP1, Cidea, Prdm16, Pgc1α, Dio2, Zic1, Fgf21, P2rx5, and PPARα(P<0.05). Also, maternal intake of EPA/ DHA enriched diet enhanced browning of sWAT by stimulating the expression of beige markers such as UCP1, Shox2, Tmem26, and PAT2 and thermogenesis regulating genes (PPARα, PGC1α, Cox7α1, and Cox8β) (P<0.05). Taken together, maternal ingestion of EPA and DHA may be suggested as a therapeutic option to encounter many pathophysiological disorders, mainly childhood obesity, and confer long-lasting metabolic benefits on offspring.

PSXIII-3 Maternal eicosapentaenoic acid feeding decreases placental and fetal lipid deposition and improves antioxidant capacity through a Sirtuin-1 independent

Abstract This research aimed to evaluate whether maternal dietary n-3 polyunsaturated fatty acids (PUFA) supplementation could decrease placental lipid deposition and improve placental antioxidant capacity through a SIRT1-dependent way. In the present study, male and female SIRT1+/- mice were mated overnight, then primiparous SIRT1+/- mice were fed a 60% kcal high-fat-diet (HFD) or equienergy EPA diet (4.4% EPA-ethyl ester) (n = 22/each group). At pregnant 18.5 d, retro-orbital blood samples were obtained then sacrificed by cervical dislocation. Placentas and its corresponding fetuses were collected by caesarean section. We found that the EPA diet significantly changed maternal plasma, placenta and fetus fatty acid composition, and decreased placental and fetal lipid content. Meanwhile, EPA treatment significantly increased placental and fetal antioxidant capacity, however, lipid peroxidation products malondialdehyde was also increased. Moreover, EPA treatment significantly activated placental UCP1 and PGC1α expression and alleviated mitochondrial swelling induced by HFD. SIRT1 knockout had no significant effect on placental lipid content, but notably decreased fetal triglyceride (TG) and non-esterified fatty acid (NEFA) content. Meanwhile, SIRT1 knockout significantly decreased the ratio of n-3 PUFA in placenta. SIRT1 knockout also significantly decreased PGC1α mRNA and protein level and increased mitochondrial swelling. Both EPA treatment and SIRT1 deficiency had no effect on BODIPY-FL-C16 (a fluorescent labeled palmitic acid) uptake in placenta and fetus. No significant effect was observed between diet and genotype. In conclusion, our study demonstrated that maternal EPA feeding decreased placental and fetal lipid deposition and improved antioxidant capacity through a Sirtuin-1 independent mechanism.

Development of New Cobalt-Free Maraging Steel with Superior Mechanical Properties via Electro-Pulsing Technology

The ductility of cobalt-free maraging steel is unsatisfactory because of the high content of Ti. Traditional heat treatment can not effectively improve the ductility of this kind steel. In this contribution, high-energy electro-pulsing is adopted in a T250 steel to solve this problem efficiently. It is found that the EPS treatment (electro-pulsing treatment under water-cooling condition) can accelerate the formation of retained austenite and nano stacking faults. Meanwhile, the microstructure is also refined by EPS treatment. Then, taking the EPS sample as the initial state, nano-reverted austenite combined with finer η-Ni3Ti precipitates are formed during EPA treatment (electro-pulsing treatment under air-cooling condition), compared with TA (traditional aging) treatment. The results of mechanical properties indicate that the strength and elongation are both enhanced by electro-pulsing treatment. Consequently, the electro-pulsing treatment can be a promising technology to devise cobalt-free maraging steel with better properties.

Eicosapentaenoic acid prevents salt sensitivity in diabetic rats and decreases oxidative stress.

Salt sensitivity (SS) is associated with increased cardiovascular risk in patients with Type 2 diabetes mellitus (T2-DM) due to an increase in renal oxidation. ω-3 polyunsaturated fatty acids have shown antioxidant effects, but a typical Western diet contains limited content. In particular, ω-3 polyunsaturated fatty acids are able to activate nuclear factor erythroid 2-related factor 2 (Nrf-2) to prevent diabetes mellitus-related complications by mitigating oxidative stress. Therefore, we hypothesized that eicosapentaenoic acid (EPA; ω-3) modulates SS in rats with T2-DM by decreasing renal oxidative stress via Nrf-2 activation and enhancing the antiinflammatory response via interleukin (IL) 6 modulation. Three-month-old male rats (n=40) were fed with a Normal Na-diet (NNaD) and randomly selected into four groups: Healthy Wistar nondiabetic rats (Wi), diabetic controls (eSS), arachidonic acid-treated eSS (AA; ω-6), and EPA-treated eSS (ω-3). After 1 year, rats were placed in metabolic cages for 7 d and fed a NNaD, followed by a 7-d period with a High Na-diet (HNaD). Systolic blood pressure, body weight, serum IL-6 and reactive oxygen species (ROS) levels were determined at the end of each 7-d period. Glycated hemoglobin (HbA1c), triacylglycerol, creatinine, and cholesterol levels were determined. ROS levels and Nrf-2 expression in kidney lysates were also assayed. Histologic changes were evaluated. A t test or analysis of variance was used for the statistical analysis. After a HNaD, systolic blood pressure increased in both the control eSS and AA groups, but not in the EPA and Wi groups. However, HbA1c levels remained unchanged by the treatments, which suggests that the observed beneficial effect was independent of HbA1c levels. The IL-6 levels were higher in the eSS and AA groups, but remained unaltered in EPA and Wi rats after a HNaD diet. Interestingly, EPA protected against serum ROS in rats fed the HNaD, whereas AA did not. In kidney lysates, ROS decreased significantly in the EPA group compared with the eSS group, and Nrf-2 expression was consistently higher compared with the AA and eSS groups. Diabetic rats presented focal segmental sclerosis, adherence to Bowman capsule, and mild-to-moderate interstitial fibrosis. EPA and AA treatment prevented kidney damage. An adequate ω3-to-ω6 ratio prevents SS in diabetic rats by a mechanism that is independent of glucose metabolism but associated with the prevention of renal oxidative stress generation. These data suggest that EPA antioxidant properties may prevent the development of hypertension or kidney damage.

Clinical Efficacy and Biological Regulations of ω–3 PUFA-Derived Endocannabinoids in Major Depressive Disorder

Background: Endocannabinoids (ECs) are one type of bioactive endogenous neuroinflammatory mediator derived from polyunsaturated fatty acids (PUFAs), which may regulate the emotional processes. Here, we assessed the effect of ω–3 PUFAs on EC levels, which may be the novel targets for the ω–3 PUFAs’ antidepressive effects. Methods: We conducted a 12-week double-blind, nonplacebo, randomized controlled trial. Eighty-eight major depressive disorder (MDD) participants were randomly assigned to receive eicosapentaenoic acid (EPA, 3.0 g/day), docosahexaenoic acid (DHA, 1.4 g/day), or a combination of EPA (1.5 g/d) and DHA (0.7 g/day). Eighty-five participants completed the trial, and their clinical remission and plasma PUFA-derived EC levels (pmol/mL) were measured. Results: The cumulative rates of clinical remission were significantly higher in the EPA and EPA + DHA groups than the DHA group (51.85 and 53.84 vs. 34.37%; p =0.027 and p =0.024, respectively). EPA and EPA + DHA treatments increased the eicosapentaenoylethanolamide (EPEA) levels compared to DHA treatment (0.33 ± 0.18 and 0.35 ± 0.24 vs. 0.08 ± 0.12; p =0.002 and p =0.001, respectively), while EPA + DHA treatment increased the docosahexaenoylethanolamide levels more than EPA treatment (1.34 ± 2.09 vs. 0.01 ± 1.79; p =0.006). Plasma EPEA levels were positively correlated with rates of clinical remission (hazard ratio: 1.60, 95% confidence interval: 1.08–2.39). Conclusions: Treatments enriched with EPA increased plasma EPEA levels, which was positively associated with clinical remission. This finding may suggest that levels of plasma EPEA play a potential novel endogenous therapeutic target in MDD.

Effect of emodin on Ang II/βig-h3 signaling pathway in diabetic nephropathy rats kidney

Salt sensitivity (SS) is associated with increased cardiovascular risk in patients with Type 2 diabetes mellitus (T2-DM) due to an increase in renal oxidation. ω-3 polyunsaturated fatty acids have shown antioxidant effects, but a typical Western diet contains limited content. In particular, ω-3 polyunsaturated fatty acids are able to activate nuclear factor erythroid 2-related factor 2 (Nrf-2) to prevent diabetes mellitus–related complications by mitigating oxidative stress. Therefore, we hypothesized that eicosapentaenoic acid (EPA; ω-3) modulates SS in rats with T2-DM by decreasing renal oxidative stress via Nrf-2 activation and enhancing the antiinflammatory response via interleukin (IL) 6 modulation.Three-month-old male rats (n = 40) were fed with a Normal Na-diet (NNaD) and randomly selected into four groups: Healthy Wistar nondiabetic rats (Wi), diabetic controls (eSS), arachidonic acid-treated eSS (AA; ω-6), and EPA-treated eSS (ω-3). After 1 year, rats were placed in metabolic cages for 7 d and fed a NNaD, followed by a 7-d period with a High Na-diet (HNaD). Systolic blood pressure, body weight, serum IL-6 and reactive oxygen species (ROS) levels were determined at the end of each 7-d period. Glycated hemoglobin (HbA1c), triacylglycerol, creatinine, and cholesterol levels were determined. ROS levels and Nrf-2 expression in kidney lysates were also assayed. Histologic changes were evaluated. A t test or analysis of variance was used for the statistical analysis.After a HNaD, systolic blood pressure increased in both the control eSS and AA groups, but not in the EPA and Wi groups. However, HbA1c levels remained unchanged by the treatments, which suggests that the observed beneficial effect was independent of HbA1c levels. The IL-6 levels were higher in the eSS and AA groups, but remained unaltered in EPA and Wi rats after a HNaD diet. Interestingly, EPA protected against serum ROS in rats fed the HNaD, whereas AA did not. In kidney lysates, ROS decreased significantly in the EPA group compared with the eSS group, and Nrf-2 expression was consistently higher compared with the AA and eSS groups. Diabetic rats presented focal segmental sclerosis, adherence to Bowman capsule, and mild-to-moderate interstitial fibrosis. EPA and AA treatment prevented kidney damage.An adequate ω3-to-ω6 ratio prevents SS in diabetic rats by a mechanism that is independent of glucose metabolism but associated with the prevention of renal oxidative stress generation. These data suggest that EPA antioxidant properties may prevent the development of hypertension or kidney damage.

A novel strengthening and toughening strategy for T250 maraging steel: Cluster-orientation governed higher strength-ductility combination induced by electropulsing

T250 steel is promising maraging steel for the development of ultrahigh strength structural materials due to the reduced cost of losing Co element. Whereas, the ductility of this steel is unsatisfactory owing to its high content of Ti. In this research, high-energy and instantaneous electropulsing is devised to improve the ductility of T250 steel. What's more, the substructure evolution and aging behavior of T250 steel after S (solution) + TA (traditional aging) and S + EPA (electropulsing treated followed with air cooling) treatment are systematically investigated by Transmission Electron Microscope (TEM) characterization, Atom Probe Technology (APT), Finite Element Modelling (FEM) simulation and Electron Back-Scattered Diffraction (EBSD) technology. It is found that unlike S + TA sample, the S + EPA sample consists of Ni3(Ti,Al) clusters, Ni2.67Ti1.33 phase and NiTi phase, respectively, which have different morphologies and sizes caused by their diverse enthalpies of formation and the thermal effect induced by the different conductivity. Furthermore, a great quantity of sub-grain boundaries, relatively considerable α variants and higher schmid factors are detected in S + EPA sample. Consequently, the higher ultimate strength (1855 MPa, 4.4% higher than S + TA sample) with better ductility (15.1%, 23.7% higher than S + TA sample) is obtained.

Eicosapentaenoic acid ethyl ester improves endothelial dysfunction in type 2 diabetic mice

BackgroundEicosapentaenoic acid (EPA) is thought to have many beneficial effects, such as anti-atherosclerogenic and anti-inflammatory properties. However, few studies have reported its effects of endothelial dysfunction in diabetes and its direct effects on the aorta. Here, we investigated the effects of EPA treatment on impaired endothelium-dependent relaxation of the aorta in KKAy mice, a model of type 2 diabetes.MethodsMale KKAy mice were fed a high-fat (HF) diet for 8 weeks to induce diabetes, after which they were divided into two groups. One group was fed a HF diet, and the other group was fed a HF diet containing EPA ethyl ester (EPA-E, 10 mg/day) for 4 weeks. Then, the vascular reactivities of prepared aortic rings were measured in an organ bath to determine if EPA-E administration changed vascular function in these diabetic mice. In addition, we examined effect of EPA-E and its metabolites to vascular action using aorta separated from C57BL/6 J mice.ResultsAlthough EPA-E administration did not change the plasma glucose and insulin levels in diabetic mice, total cholesterol levels were significantly decreased. The aorta extracted from EPA-E untreated diabetic mice showed impaired endothelium-dependent relaxation in response to acetylcholine (ACh). However, EPA-E administration improved the relaxation response to ACh to the control levels observed in non-diabetic C57BL/6 J mice. On the other hand, endothelium-independent relaxation in response to sodium nitroprusside did not significantly differ among these three groups. The enhanced contractile response by phenylephrine in diabetic mice was not altered by the administration of EPA-E. In addition, the direct administration of EPA-E metabolites such as EPA, docosahexaenoic acid, and docosapentaenoic acid led to vasodilation in the aortic rings of C57BL/6 J mice.ConclusionThese results showed that chronic EPA-E administration prevented the development of endothelial dysfunction in KKAy mice, partly via the direct action of EPA-E metabolites on the aorta.

The impact of polyunsaturated fatty acids on DNA methylation and expression of DNMTs in human colorectal cancer cells

Growing evidence suggests a role of polyunsaturated fatty acids (PUFA) in the prevention of various types of malignancy, including colorectal cancer (CRC). No published studies have yet examined the direct effect of PUFA treatment on DNA methylation in CRC cells. In this study, 5 human CRC cells were treated with 100 μM DHA, EPA, and LA for 6 days and changes in their global- and gene-specific DNA methylation status as well as expression of DNA methyl transferases (DNMT) were investigated. Cell-type specific differences in DNA methylation and expression of DNMTs were observed in PUFA-treated cells. DHA and EPA treatment induced global hypermethylation in HT29/219 and HCT116 cells, but reduced methylation in Caco2 cells (p < 0.05). Among 10 tumor related genes tested in 5 CRC cell lines, DHA and EPA induced promoter demethylation of Cox2 in HT29/219, p14 and PPARγ in HCT116, and ECAD in SW742 cells. Cell-type specific differences in expression of DNMT1, DNMT3a, and 3b genes were also observed between PUFA-treated and control cells (p < 0.05). Overall, treatment of PUFAs coordinately induced the expression of DNMTs in HT29/219, but suppressed in other 4 cell lines investigated in this study.

Dietary Fatty Acids Differentially Regulate Secretion of Adiponectin and Interleukin-6 in Primary Canine Adipose Tissue Culture.

The aim of this study was to determine the effect of n3 polyunsaturated fatty acids (PUFA) on canine adipose tissue secretion of adiponectin, interleukin-6 (IL6), and tumor necrosis factor-α (TNFα). Subcutaneous and omental visceral adipose tissue samples were collected from 16 healthy intact female dogs. Concentrations of adiponectin were measured in mature adipocyte cultures, and concentrations of IL6 and TNFα were measured in undifferentiated stromovascular cell (SVC) cultures following treatment with eicosapentaenic acid (EPA, 20:5n-3), arachidonic acid (ARA, 20:4n-6), or palmitic acid (PAM, 16:0) at 25, 50, or 100 μM. Secretion of adiponectin from mature adipocytes was higher (p < 0.001) following EPA treatment at 50 μM compared to control in subcutaneous tissue, and higher following EPA treatment compared to PAM treatment at 25 μM in both subcutaneous (p < 0.001) and visceral tissues (p = 0.010). Secretion of IL6 from SVC derived from subcutaneous tissue was lower following EPA treatment and higher following PAM treatment compared to control both at 50 μM (p = 0.001 and p = 0.041, respectively) and 100 μM (p = 0.013 and p < 0.001, respectively). These findings of stimulation of adiponectin secretion and inhibition of IL6 secretion by EPA, and stimulation of IL6 secretion by PAM, are consistent with findings of increased circulating concentrations of adiponectin and decreased circulating concentration of IL6 in dogs supplemented with dietary fish oil, and show that the effect of fish oil on circulating concentrations of adiponectin and IL6 is, at least partially, the result of local effects of EPA and PAM on adipose tissue.

Abstract TP429: Association Between Perioperative Management and Outcome in Aged SAH Patients

Background and purpose: The outcomes of subarachnoid hemorrhage (SAH) in aged patients are more severe than those in non-aged patients. There are few reports about the relationship between the age and the effect of perioperative care for SAH patients. We performed a nationwide survey in Japan to determine the relationship between perioperative care and SAH outcomes in aged and non-aged patients. Methods: We analyzed 17,343 subarachnoid hemorrhage (SAH) patients treated with clipping or coiling in 579 hospitals who participated in the J-ASPECT study. Data between 2010 and 2013 were obtained from the Japanese Diagnosis Procedure Combination Database. We stratified patients into two groups according to their age (aged group &gt;75 y.o., n=3885; non-aged group &lt; 75 y.o., n=13,458) and analyzed the association between perioperative care and poor outcome (modified Rankin Scale score 3-6 at the time of discharge). With respect to perioperative care, we evaluated time from onset to surgery (days), treatment (clipping or coiling), and drugs delivered after surgery (fasudil hydrochloride, ozagrel sodium, cilostazol, statin, EPA, edaravone). Results: In the non-aged group, coiling (OR=0.84; P&lt;0.01) and treatment with fasudil hydrochloride (OR=0.59; P&lt;0.01), statin (OR=0.83; P&lt;0.01), and EPA (OR=0.83; P&lt;0.01) significantly improved the outcomes and cilostazol treatment tended to improve the outcomes (OR=0.91; P=0.07). In both groups, edaravone treatment was associated with poor outcome (aged group: OR=2.34, P&lt;0.01; non-aged group: OR=2.33, P&lt;0.01). Although no factor that could improve outcome in the aged group was identified, JCS scores less than 30, coiling (OR=0.80; P=0.03), and EPA treatment (OR=0.74; P=0.02) were statistically significant prognosis improvement factors. Conclusion: Coiling and treatment with fasudil hydrochloride, statins, and EPA improved outcomes of non-aged patients. Although perioperative care did not improve the outcome of aged SAH patients, in cases of relatively mild SAH, perioperative care had the potential to improve the outcome.

Krill oil extract suppresses cell growth and induces apoptosis of human colorectal cancer cells

BackgroundColorectal cancer (CRC) is the third most common cancer in the world. The current available treatments for CRC include surgery, chemotherapy and radiotherapy. However, surgery is only useful when the disease is diagnosed at the earlier stage. Chemotherapy and radiotherapy are associated with numerous side effects that decrease the patients’ quality of life. Safer, effective alternatives, such as natural compounds, to chemotherapy are desirable. This study assessed the efficacy of free fatty acid (FFA) extract of krill oil on three human CRC cells lines.MethodsHCT-15, SW-480 and Caco-2 cells were treated with the FFA extracts of krill oil and fish oil for 48 h while treatments with the bioactive omega-3 polyunsaturated fatty acids (LC n-3 PUFA) of these marine oils, eicosapentaenoic acid (EPA, C20:5n-3) and docosahexaenoic acid (DHA, C22:6n-3) in comparison with a n-6 PUFA, arachnoid acid (AA, C20:4n-6) were up to 72 h at the concentrations of 50, 100, 150 and 200 μM. Effects of all the treatments on cell proliferation were assessed using a water-soluble tetrazolium-1 (WST-1) assay kit at 24, 48 and 72 h. Effects of FFA extract of krill oil and EPA on apoptosis and mitochondrial membrane potential were determined using commercial kits after 48 h of treatment.ResultsKrill oil extract inhibited cell proliferation of all three cell lines in the similar manner as fish oil extract. A significant cell apoptosis and increase in mitochondrial membrane potential were observed after the treatment with krill oil extract. EPA at the concentration of 200 μM reduced significantly the proliferation of HCT-15 and SW-480 at 24, 48 and 72 h. In addition, EPA treatment (100 and 200 μM) resulted in significant cell apoptosis in all three cell lines. No significant changes were observed after treatment with DHA and AA.ConclusionsOur results indicate that the FFA extract of krill oil maybe an effective chemotherapeutic agent to suppress proliferation and induce apoptosis in CRC cells through its bioactive constitute EPA. Although the exact mechanism of the pro-apoptotic properties of krill oil extract is unclear, mitochondrial pathway seems to be implicated.

Membrane Disordering by Eicosapentaenoic Acid in B Lymphomas Is Reduced by Elongation to Docosapentaenoic Acid as Revealed with Solid-State Nuclear Magnetic Resonance Spectroscopy of Model Membranes

Background: Plasma membrane organization is a mechanistic target of n–3 (ω-3) polyunsaturated fatty acids. Previous studies show that eicosapentaenoic acid (EPA; 20:5n–3) and docosahexaenoic acid (DHA; 22:6n–3) differentially disrupt plasma membrane molecular order to enhance the frequency and function of B lymphocytes. However, it is not known whether EPA and DHA affect the plasma membrane organization of B lymphomas differently to influence their function.Objective: We tested whether EPA and DHA had different effects on membrane order in B lymphomas and liposomes and studied their effects on B-lymphoma growth.Methods: B lymphomas were treated with 25 μmol EPA, DHA, or serum albumin control/L for 24 h. Membrane order was measured with fluorescence polarization, and cellular fatty acids (FAs) were analyzed with GC. Growth was quantified with a viability assay. 2H nuclear magnetic resonance (NMR) studies were conducted on deuterated phospholipid bilayers.Results: Treating Raji, Ramos, and RPMI lymphomas for 24 h with 25 μmol EPA or DHA/L lowered plasma membrane order by 10–40% relative to the control. There were no differences between EPA and DHA on membrane order for the 3 cell lines. FA analyses revealed complex changes in response to EPA or DHA treatment and a large fraction of EPA was converted to docosapentaenoic acid (DPA; 22:5n–3). NMR studies, which were used to understand why EPA and DHA had similiar membrane effects, showed that phospholipids containing DPA, similar to DHA, were more ordered than those containing EPA. Finally, treating B lymphomas with 25 μmol EPA or DHA/L did not increase the frequency of B lymphomas compared with controls.Conclusions: The results establish that 25 μmol EPA and DHA/L equally disrupt membrane order and do not promote B lymphoma growth. The data open a new area of investigation, which is how EPA’s conversion to DPA substantially moderates its influence on membrane properties.

Abstract 243: Regression of Carotid Intima-media Thickness and Improvement of Lipid Profile by the Addition of High-purity Eicosapentaenoic Acid (Icosapent Ethyl) to Statins in a Morbidly Obese Patient with Type II Diabetes Mellitus

Objective: The AHA scientific statement on obesity identifies obesity as an important independent metabolic/genetic risk factor for ASCVD. For patients at high risk of ASCVD and with less-than-anticipated therapeutic response to statin therapy, the 2013 ACC/AHA cholesterol guideline recommends increasing statin intensity, or if on maximally tolerated statin intensity, considering non-statin therapy. The 2013 ACC/AHA cholesterol guideline evidence statements also describe the JELIS trial, in which 1.8 g/day of EPA added to statin therapy led to a 19% relative risk reduction in major coronary events in Japanese patients with hypercholesterolemia. Additionally, in the CHERRY trial of high-risk cardiac patients treated with moderate- to high-intensity statin therapy, the addition of 1.8 g/day of EPA resulted in significantly greater prevalence of coronary plaque regression compared to statin therapy alone (50% vs 24%; p&lt;0.001). Further, in patients with DM, ASCVD is the primary cause of death. Methods: We reviewed the impact of initiating a prescription high-purity EPA drug, icosapent ethyl (2 g/day), in a morbidly obese (485 lb) 51-year-old African-American male patient with hypertension, DM, and ASCVD (evidenced by high CIMT) while on maximally tolerated statin therapy. Diet, exercise, and medications were stable for the duration of this report. Lipids, A1C, HOMA-IR, and CIMT were evaluated before and 4 months after initiating icosapent ethyl. Results: See table. Conclusions: Adding high-purity prescription EPA treatment (icosapent ethyl) to a maximally tolerated statin dose in a morbidly obese African-American patient with DM led to substantial regression of carotid intima-media thickness and to improvements in measured lipid parameters and two laboratory measures of DM. The clinical implications of adding icosapent ethyl to statin therapy is being evaluated in the ongoing cardiovascular outcomes trial, REDUCE-IT.

EARLY INITIATION OF EICOSAPENTAENOIC ACID AND STATIN TREATMENT IS ASSOCIATED WITH BETTER CLINICAL OUTCOMES THAN STATIN ALONE IN PATIENTS WITH ACUTE CORONARY SYNDROMES: 1-YEAR OUTCOMES OF A RANDOMIZED CONTROLLED STUDY

Early initiation of EPA treatment in combination with a statin within 24 h after percutaneous coronary intervention (PCI) in patients with acute myocardial infarction (MI) reduces inflammation and ventricular arrhythmia compared with statin monotherapy; however, the impact of early initiation of EPA