The Investigation of the Effects of Eicosapentaenoic Acid and Docosahexaenoic Acid on Cultured Human Airway Epithelial Cells
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3 Abstract: Fatty acid composition has an important role in cellular structure and function. Inflammatory behavior of the airway epithelial cells can be changed due to the manipulation of their fatty acid content, which has a critical importance in asthma. The objective of the present study was to determine the fatty acid composition of human airway epithelial cells after co-culturing with eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Hence, airway epithelial cells (Calu-3, Passage 40-43, from ATCC, USA) were incubated with different concentrations (0, 10, 200, 400µM) of EPA and DHA for 24h at 37C in the presence of 5% CO and the 2 incorporation of fatty acids was analyzed using gas chromatography (GC). Findings showed that there was a significant decreasing trend in the concentration of n-6 polyunsaturated fatty acids (PUFAs) and non- significant increasing trend in total n-3 PUFAs due to EPA and DHA supplementation. As a result of EPA incorporation, the levels of saturated fatty acids (SFAs), monounsaturated fatty acids (MUFAs) and DHA declined significantly. On the contrary, docosapentaenoic acid (DPA) content elevated markedly due to EPA supplementation. The EPA concentration increased significantly as a consequence of DHA incorporation. Additionally, the n-3/n-6 ratio elevated notably in both DHA and EPA supplemented groups. In conclusion, incorporation of DHA and EPA can alter the fatty acid content of airway epithelial cells in a way which has a lower inflammatory characteristic.Keywords:
Docosapentaenoic acid
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Abstract 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.
Phosphatidylethanolamine
Lipidology
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Treatment with high dose icosapent ethyl (IPE), an ethyl ester of the omega-3 fatty acid eicosapentaenoic acid (EPA), significantly reduced ischemic events in patients with either cardiovascular disease (CV) or diabetes plus other risk factors (REDUCE-IT) but the mechanism is not well understood. We compared the effects of EPA, docosahexaenoic acid (DHA), and the omega-6 fatty acid arachidonic acid (AA) on bioavailability of nitric oxide (NO) and fatty acid composition. Human umbilical vein endothelial cells (HUVECs) were pretreated with EPA, DHA, or AA (10 µM). Cells were stimulated with calcium ionophore and NO and peroxynitrite (ONOO-) were measured using porphyrinic nanosensors. Levels of EPA, DHA, AA and other fatty acids were measured by gas chromatography (GC). EPA treatment caused the greatest NO release (18%, p < 0.001) and reduction in ONOO- (13%, p < 0.05) compared to control; the [NO]/[ ONOO-] ratio increased by 35% (p < 0.001). DHA treatment increased NO levels by 12% (p < 0.01) but had no effect on ONOO- release. AA did not affect either NO or ONOO- release. Fatty acid treatments increased their respective levels in endothelial cells. EPA levels increased 10-fold to 4.59 mg/g protein (p < 0.001) with EPA treatment and the EPA/AA ratio increased by 10-fold (p < 0.001) compared to vehicle. Only EPA increased docosapentaenoic acid (DPA, omega-3) levels by 2-fold (p < 0.001). AA alone decreased the EPA/AA ratio 4-fold (p<0.001). These findings support a preferential benefit of EPA on endothelial function and omega-3 fatty acid content.
Docosapentaenoic acid
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Docosahexaenoic acid (DHA) plays an important role in visual and neural development in mammals. In the present study, effect of dietary supplementation with n-3 fatty acids, primarily docosahexaenoic acid (DHA) with high purity, on the fatty acid composition of photoreceptor cells of young rats (fed from 4 weeks) was investigated. DHA in rod outer segment (ROS) membranes was significantly increased in the group of high DHA feeding (9.69% total energy). Other n-3 fatty acids (α-linolenic acid (ALA) and eicosapentaenoic acid (EPA)) included in the diets with DHA (0.95%∼5.63% total energy) also significantly increased the proportion of DHA compared with the linoleic acid diet groups. However, the proportions of arachidonic acid (ARA) and other long chain n-6 fatty acids (22:4n6 and 22:5n6) were suppressed in these n-3 fatty acids-fed groups. Phospholipid hydroperoxides in ROS membranes were determined using a highly sensitive analytical technique, chemiluminescence-high performance liquid chromatography (CL-HPLC). There was no increasing tendency in the hydroperoxide levels of ROS membranes containing high content of DHA, and phosphatidylethanolamine hydroperoxide (PEOOH) was much lower than phosphatidylcholine hydroperoxide (PCOOH) under normal light conditions, which implies that DHA supplementation does not much affect the peroxidizability of ROS membranes in vivo. But UV irradiation on separated ROS membranes accelerated the formation of phospholipid hydroperoxides in high DHA feeding rats, and PEOOH was produced more efficiently than PCOOH in vitro.
Phosphatidylethanolamine
Linolenic acid
Degree of unsaturation
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Objective: Adipose tissue (AT)-specific inflammation in obesity is associated with several metabolic disorders such as insulin resistance, and hepatic steatosis. We and others have shown that fish oil containing the omega-3 fatty acids increases AT mass in animal models which is associated with reduced AT-inflammation, hepatic steatosis, and dyslipidemia. Therefore, the objective of this study is to determine whether the omega-3 fatty acids such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) induce the differentiation process in 3T3-L1 preadipocytes. Methods: 3T3-L1 cells were pretreated and differentiated in the presence of different long chain fatty acids such as oleic acid (OA), stearic acid (SA), EPA, DHA, or a mixture of EPA and DHA (E+D) at a concentration of 50 μM. Results: Oil red O staining revealed that the differentiation process was remarkably increased in cells treated with DHA and, to a lesser extent with SA, compared to control cells differentiated in the absence of fatty aci...
Oil Red O
Steatosis
Dyslipidemia
3T3-L1
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Docosapentaenoic acid
3T3-L1
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Epidemiological studies suggest that high fish intake is associated with a decreased risk of colorectal cancer which has been linked to the high content of the n - 3 polyunsaturated fatty acids (PUFAs) eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in some fish. In this study, two different cell lines are compared in relation to their response to EPA and DHA versus the plant derived PUFAs, linoleic acid (LA), gamma-linolenic acid (GLA), and alpha-linolenic acid (ALA) and to the ubiquitous arachidonic acid (ARA). The uptake of 100 microM of each fatty acid (FA) was determined using GC. The 4',6-diamidino-2-phenylindole assay for DNA quantification and the Cell-Titer-Blue assay were used to determine cell survival and metabolic activity at 2-72 h after treatment. All FAs were utilized more efficiently by the human colon adenoma cell line LT97 than by the adenocarcinoma cell line HT29. LT97 were more susceptible than HT29 cells to the growth inhibitory activities of all FAs except for DHA where both were equally sensitive. Inhibition of survival and metabolic activity by EPA and DHA increased with treatment time in both cell lines. ALA or GLA were less growth inhibitory than EPA or DHA and ARA had intermediary activity. The data show that the tested FAs are incorporated into colon cells. Furthermore, adenoma cells are more susceptible than the adenocarcinoma cells.
Linolenic acid
alpha-Linolenic acid
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Abstract Previous studies in our laboratory have shown that marine oils, with high levels of eicosapentaenoic (EPA, 20∶5n−3) and docosahexaenoic acids (DHA, 22∶6n−3), inhibit the growth of CT‐26, a murine colon carcinoma cell line, when implanted into the colons of male BALB/c mice. An in vitro model was developed to study the incorporation of polyunsaturated fatty acids (PUFA) into CT‐26 cells in culture. PUFA‐induced changes in the phospholipid fatty acid composition and the affinity with which different fatty acids enter the various phospholipid species and subspecies were examined. We found that supplementation of cultured CT‐26 cells with either 50 μM linoleic acid (LIN, 18∶2n−6), arachidonic acid (AA, 20∶4n−6), EPA, or DHA significantly alters the fatty acid composition of CT‐26 cells. Incorporation of these fatty acids resulted in decreased levels of monounsaturated fatty acids, while EPA and DHA also resulted in lower levels of AA. While significant elongation of both AA and EPA occurred, LIN remained relatively unmodified. Incorporation of radiolabeled fatty acids into different phospholipid species varied significantly. LIN was incorporated predominantly into phosphatidylcholine and had a much lower affinity for the ethanolamine phospholipids. DHA had a higher affinity for plasmenylethanolamine (1‐ O ‐alk‐1′‐enyl‐2‐acyl‐ sn ‐glycero‐3‐phosphoethanolamine) than the other fatty acids, while EPA had the highest affinity for phosphatidylethanol‐amine (1,2‐diacyl‐ sn ‐glycero‐3‐phosphoethanolamine). These results demonstrate that, in vitro , significant differences are seen between the various PUFA in CT‐26 cells with respect to metabolism and distribution, and these may help to explain differences observed with respect to their effects on tumor growth and metastasis in the transplantable model.
Lipidology
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Fatty Acid Metabolism
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CYP2C8
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