Δ6 Desaturase Activity Research Articles

Total dietary fat and fatty acid content modifies plasma phospholipid fatty acids, desaturase activity indices, and urinary prostaglandin E in women

Compared with diets high in fat, low-fat diets are associated with reduced risk of cardiovascular disease. We hypothesized that a low-fat (LF) (20% fat) and an LF high-omega-3 (n-3) fatty acid diet (LFn3) (23% fat with 3% as α-linolenic acid, eicosapentaenoic acid, and docosahexaenoic acid [DHA]) would enhance n-3 composition of plasma phospholipid fatty acid and reduce urinary prostaglandin E(2) (PGE(2)) relative to a high-fat diet (HF) (40% fat) and that these changes would be associated with alterations in δ5 desaturase (D5D) and δ6 desaturase (D6D) activity. Phospholipid fatty acids and urinary PGE(2) were measured, and D5D and D6D activity indices calculated in a crossover trial in 17 postmenopausal women fed each of 3 test diets (HF, LF, and LFn3) for 8-week feeding periods. Desaturase activity indices were calculated as D5D, 20:4n-6/20:3n-6, and D6D, 20:3n-6/18:2n-6. Plasma phospholipid fatty acid, α-linolenic acid, eicosapentaenoic acid, docosapentaenoic acid (DPA), DHA, and total n-3 fatty acids increased, whereas linoleic acid and arachidonic acid decreased with consumption of LFn3. The LF resulted in enhanced arachidonic acid and DHA. High fat reduced D6D, whereas both HF and LF increased D5D. Urinary PGE(2) was reduced in response to both the LF and LFn3 diets. Low-fat diets, with or without long-chain n-3 fatty acids, promote positive health effects due in part to favorable alteration of plasma phospholipid fatty acid profiles and modification in desaturase activity indices, suggesting that the type and amount of fat consumed are modifiable risk factors for the prevention of cardiovascular disease.

Effect of maternal micronutrients (folic acid, vitamin B 12) and omega 3 fatty acids on liver fatty acid desaturases and transport proteins in Wistar rats

A disturbed fatty acid metabolism increases the risk of adult non-communicable diseases. This study examines the effect of maternal micronutrients on the fatty acid composition, desaturase activity, mRNA levels of fatty acid desaturases and transport proteins in the liver. Pregnant female rats were divided into 6 groups at 2 levels of folic acid both in the presence and absence of vitamin B 12. The vitamin B 12 deficient groups were supplemented with omega 3 fatty acid. An imbalance of maternal micronutrients reduces liver docosahexaenoic acid, increases Δ5 desaturase activity but decreases mRNA levels, decreases Δ6 desaturase activity but not mRNA levels as compared to control. mRNA level of Δ5 desaturase reverts back to the levels of the control group as a result of omega 3 fatty acid supplementation. Our data for the first time indicates that maternal micronutrients differentially alter the activity and expression of fatty acid desaturases in the liver.

Effect of tanniniferous Terminalia chebula extract on rumen biohydrogenation, ∆9-desaturase activity, CLA content and fatty acid composition in longissimus dorsi muscle of kids

Conjugated linoleic acid, a fatty acid found in milk fat and ruminant meat is one of the functional food components. Modifying fatty acid composition so as to increase CLA and other beneficial PUFA/MUFA level and reducing SFA levels might be a key to enhance the neutraceutical and therapeutic value of ruminant-derived food products. In the present experiment, the effect of supplementation of polyphenol rich Terminalia chebula plant extract at different concentrations (1.06g/kg and 3.18g/kg of body weight in T1 and T2 groups, respectively) was investigated on fatty acid composition of rumen fluid, plasma, intramuscular fat and Δ9-desaturase activity in longissimus dorsi muscle of crossbred kids. Total MUFA and PUFA content in muscle were enhanced by 25 and 35%, respectively, whereas SFA was reduced by 20% thereby improving the desaturation index. Δ9-desaturase activity also increased by 47% resulting in an enhancement of total CLA content (58.73%) in muscle.

High level accumulation of gamma linolenic acid (C18:3Δ6.9,12 cis) in transgenic safflower (Carthamus tinctorius) seeds

Gamma linolenic acid (GLA; C18:3Δ6,9,12 cis), also known as γ-Linolenic acid, is an important essential fatty acid precursor for the synthesis of very long chain polyunsaturated fatty acids and important pathways involved in human health. GLA is synthesized from linoleic acid (LA; C18:2Δ9,12 cis) by endoplasmic reticulum associated Δ6-desaturase activity. Currently sources of GLA are limited to a small number of plant species with poor agronomic properties, and therefore an economical and abundant commercial source of GLA in an existing crop is highly desirable. To this end, the seed oil of a high LA cultivated species of safflower (Carthamus tinctorius) was modified by transformation with Δ6-desaturase from Saprolegnia diclina resulting in levels exceeding 70% (v/v) of GLA. Levels around 50% (v/v) of GLA in seed oil was achieved when Δ12-/Δ6-desaturases from Mortierella alpina was over-expressed in safflower cultivars with either a high LA or high oleic (OA; C18:1Δ9 cis) background. The differences in the overall levels of GLA suggest the accumulation of the novel fatty acid was not limited by a lack of incorporation into the triacylgylcerol backbone (>66% GLA achieved), or correlated with gene dosage (GLA levels independent of gene copy number), but rather reflected the differences in Δ6-desaturase activity from the two sources. To date, these represent the highest accumulation levels of a newly introduced fatty acid in a transgenic crop. Events from these studies have been propagated and recently received FDA approval for commercialization as Sonova™400.

ELOVL2 controls the level of n-6 28:5 and 30:5 fatty acids in testis, a prerequisite for male fertility and sperm maturation in mice

ELOVL2 is a member of the mammalian microsomal ELOVL fatty acid enzyme family, involved in the elongation of very long-chain fatty acids including PUFAs required for various cellular functions in mammals. Here, we used ELOVL2-ablated (Elovl2(-/-)) mice to show that the PUFAs with 24-30 carbon atoms of the ω-6 family in testis are indispensable for normal sperm formation and fertility in male mice. The lack of Elovl2 was associated with a complete arrest of spermatogenesis, with seminiferous tubules displaying only spermatogonia and primary spermatocytes without further germinal cells. Furthermore, based on acyl-CoA profiling, heterozygous Elovl2(+/-) male mice exhibited haploinsufficiency, with reduced levels of C28:5 and C30:5n-6 PUFAs, which gave rise to impaired formation and function of haploid spermatides. These new insights reveal a novel mechanism involving ELOVL2-derived PUFAs in mammals and previously unrecognized roles for C28 and C30 n-6 PUFAs in male fertility. In accordance with the function suggested for ELOVL2, the Elovl2(-/-) mice show distorted levels of serum C20 and C22 PUFAs from both the n-3 and the n-6 series. However, dietary supplementation with C22:6n-3 could not restore male fertility to Elovl2(+/-) mice, suggesting that the changes in n-6 fatty acid composition seen in the testis of the Elovl2(+/-) mice, cannot be compensated by increased C22:6n-3 content.

Effect of pasture and soybean supplementation on fatty acid profile and CLA content in dairy cow milk

Conjugated linoleic acid (CLA) has been related to several beneficial effects on human and animal health (anticancerogens, antiatherogenics, etc.) The main dietary source of CLA for humans are foods derived from ruminants. In dairy products CLA content depends on ruminal bio-hydrogenation of the dietary unsaturated fats and it is also related to the Δ9desaturase activity in the mammary gland. The CLA level of milk is affected by various factors, including diet composition, that seems to play an important role (Jensen, 2002). The aim of this study was to investigate the effect of toasted and raw soybean addition to a mixed diet and

Erythrocyte membrane phospholipid fatty acids, desaturase activity, and dietary fatty acids in relation to risk of type 2 diabetes in the European Prospective Investigation into Cancer and Nutrition (EPIC)–Potsdam Study

The long-term role of fatty acids (FAs) in the cause of diabetes remains largely unclear. We aimed to investigate erythrocyte membrane FAs, desaturase activity, and dietary FAs in relation to the incidence of type 2 diabetes. We applied a nested case-cohort design (n = 2724, including 673 incident diabetes cases) within the European Prospective Investigation into Cancer and Nutrition (EPIC)-Potsdam Study, which involves 27,548 middle-aged subjects. Thirty erythrocyte membrane FAs (percentage of total FAs) and FA intake (percentage of total fat) were measured at baseline, and physician-confirmed incident diabetes was assessed during a mean follow-up of 7.0 y. We evaluated Δ⁵ desaturase (D5D) and Δ⁶ desaturase (D6D) activity by using FA product-to-precursor ratios (traditional approach) and by investigating variants in FADS1 and FADS2 genes that encode these desaturases (Mendelian randomization approach). As a main finding, erythrocyte 16:1n-7 and 18:3n-6 and FA ratios, which reflect stearoyl coenzyme A desaturase (SCD) and D6D activity, were directly related to diabetes risk in multivariable-adjusted models [relative risks (95% CIs) comparing extreme quintiles: 16:1n-7, 2.11 (1.46, 3.05); 18:3n-6, 2.00 (1.38, 2.88); SCD, 2.61 (1.75, 3.89); and D6D, 2.46 (1.67, 3.63)], whereas the FA ratio that reflects D5D activity was inversely associated with risk [0.46 (0.31, 0.70)]. The Mendelian randomization approach corroborated the direct relation for D6D activity and tended to support the inverse relation for D5D activity. Proportions of dietary FAs showed only modest to low correlations with erythrocyte FAs and were not significantly associated with risk. The FA profile of erythrocyte membrane phospholipids and activity of desaturase enzymes are strongly linked to the incidence of type 2 diabetes.

Deposition of docosahexaenoic acid (DHA) is limited in forebrain of young obese fa/ fa Zucker rats fed a diet high in α-linolenic acid but devoid of DHA

Docosahexaenoic acid (DHA) is required for neurotransmitter synthesis and learning. Conversion of α-linolenic acid (ALA) to DHA is considered adequate to support brain function in youth, but it is unknown if brain DHA can be maintained in insulin resistant states. This study investigated brain fatty acid and desaturase activities in young insulin resistant Zucker rats on diets with and without DHA. Male fa/fa and lean rats were fed diets enriched with flaxseed (FXO, ALA: 35.5% fatty acids), menhaden (MO, DHA: 9.2%) or safflower oil (SO, linoleic acid: 54.1%) for 9 weeks, n=8 per diet per genotype. Compared to lean, the 15 week old fa/fa rats were obese (56% heavier) and insulin-resistant (>18-fold in homeostasis model assessment of insulin resistance). The forebrain of fa/fa rats had higher palmitoleic (16:1n-7) and dihomo-γ-linolenic (20:3n-6) acids, and higher Δ9, Δ6 but lower Δ5 (all P≤.006) desaturase indices than lean. The Δ9 and Δ6 desaturase indices positively, while the Δ5 negatively (all P≤.01) correlated with insulin resistance. The Δ9 desaturase index positively correlated with adiposity index. The percentage of forebrain DHA of fa/fa rats was lower ( P=.011) than lean rats when fed FXO diet while there was no difference ( P>.05) between fa/fa and lean rats fed MO or SO diet. Thus, the alterations in the fatty acid and desaturase indices in the brain were consistent inhibited forebrain synthesis of DHA in the fa/fa rats. ALA may not have potential to effectively serve as a precursor for synthesizing DHA for youth forebrain during insulin resistance since Δ5 desaturase activity is limited.

Comparison of fatty acid proportions in serum cholesteryl esters among people with different glucose tolerance status: The CoDAM study

Background and aimAltered fatty acid patterns in blood may be associated with insulin resistance and related disorders. We investigated whether serum proportions of cholesteryl fatty acids and desaturase activity are associated with glucose tolerance status and insulin resistance. Methods and resultsData were obtained from a cross-sectional study among 471 Dutch participants aged ≥40 years. Individual fatty acids in serum cholesteryl esters were determined and endogenous conversions by desaturases were estimated from product-to-precursor ratios. Proportions of fatty acids were compared among participants with normal glucose tolerance, impaired glucose metabolism and newly diagnosed type 2 diabetes. Partial Spearman correlation coefficients between fatty acids and homeostasis model assessment-insulin resistance (HOMA-IR) were calculated. Adjustments were made for lifestyle and nutritional factors.The proportions of total saturated, mono-unsaturated, trans- and poly-unsaturated fatty acids did not differ significantly between groups, but several individual fatty acids did; the proportions of C18:0 and C20:3n6 were higher, whereas those of C18:1n7 and C20:4n6 were lower in participants with type 2 diabetes compared with those with normal glucose tolerance. Activity of Δ5-desaturase, that is, ratio of C20:4n6 to C20:3n6, was lower (p < 0.01) in participants with type 2 diabetes (7.4) than with normal glucose tolerance (8.4). HOMA-IR was correlated positively with Δ9-desaturase activity (r = 0.11, p < 0.01) and inversely with Δ5-desaturase activity (r = −0.21, p < 0.01). ConclusionThe observed lower Δ5-desaturase activity in participants with type 2 diabetes and its inverse association with HOMA-IR suggest that changes in fatty-acid metabolism may play a role in the aetiology of type 2 diabetes.

Elevated Delta-6 desaturase ( FADS2) gene expression in the prefrontal cortex of patients with bipolar disorder

Although evidence suggests that a dysregulation in polyunsaturated fatty acid (PUFA) homeostasis may contribute to the pathoetiology of bipolar disorder (BD), there is currently nothing known about the expression of genes that mediate long-chain PUFA biosynthesis in BD patients. In the present study we determined FADS1 (Δ5 desaturase), FADS2 (Δ6 desaturase), HELO1 [ ELOVL5] (elongase), PEX19 (peroxisome), and SCD (stearoyl-CoA desaturase [Δ9 desaturase]) mRNA expression in the postmortem prefrontal cortex of non-psychiatric controls ( n = 12) and BD patients ( n = 12) by real-time reverse transcriptase polymerase chain reaction (RT-PCR). Changes in the activities of corresponding enzyme products were estimated from fatty acid product: precursor ratios. After correcting for multiple comparisons, FADS2 mRNA expression was significantly greater in BD patients relative to controls (+27%, p = 0.004). Indices of Δ6 desaturase activity, including 20:4/18:2 (+18%, p = 0.15) and 20:3/18:2 (+12%, p = 0.25) ratios, were numerically, but not significantly, greater in BD patients relative to controls. There were no significant group differences in FADS1 (+17%, p = 0.32), HELO1 (+4%, p = 0.81), PEX19 (−2%, p = 0.91), and SCD (+4%, p = 0.85) mRNA expression, or indices of Δ5 desaturase (+5%, 0.59), elongase (+3%, p = 0.62), and stearoyl-CoA desaturase (−11%, p = 0.10) activities. These preliminary findings demonstrate that FADS2 mRNA expression is significantly and selectively elevated in the prefrontal cortex of BD patients, and may contribute to dysregulated central PUFA biosynthesis and pro-inflammatory signaling implicated in the pathophysiology of BD.

Flaxseed supplementation improves fatty acid profile of cow milk

The objective of the study was to determine the effects of adding flaxseed or fish oil to the diet on the milk fatty acid profile of cows. The experiment was conducted in the summer of 2006 and involved 24 Friesian cows that were divided into 3 groups of 8 animals according to different type of fat supplementation: a traditional diet with no fat supplementation, a diet supplemented with whole flaxseed, and a diet supplemented with fish oil. Results suggested that whole flaxseed supplementation positively affects the milk fatty acid profile during summer. In particular, milk from cows receiving flaxseed supplementation showed a decrease in saturated fatty acid, an increase in monounsaturated fatty acid, and, together with the milk from fish oil-supplemented cows, an increase in polyunsaturated fatty acid content compared with milk from control cows. As expected, both fish oil and flaxseed supplementation increased the content of n-3 polyunsaturated fatty acids in milk fat. The increased dietary intake of C18:3 in flaxseed-supplemented cows resulted in increased levels of milk C18:1 trans-11 and increased conjugated linoleic acid C18:2 cis-9,trans-11 by Δ9-desaturase activity. Milk from flaxseed-supplemented cows together with the high conjugated linoleic acid content was characterized by low atherogenic and thrombogenic indices, suggesting that its use has less detrimental effects concerning the atherosclerosis and coronary thrombosis risk associated with the consumption of milk and dairy products. In conclusion, flaxseed supplementation improves composition and nutritional properties of milk from cows milked during times of high ambient temperature.

A comparison between Holstein-Friesian and Jersey dairy cows and their F1 hybrid on milk fatty acid composition under grazing conditions

The objective of this study was to investigate the effect of 2 breeds, Holstein and Jersey, and their F1 hybrid (Jersey × Holstein) on milk fatty acid (FA) concentrations under grazing conditions, especially conjugated linoleic acid (CLA) and n-3 polyunsaturated fatty acids because of their importance to human health. Eighty-one cows (27 per breed grouping) were allocated a predominantly perennial ryegrass pasture. Samples were collected over 2 periods (June and July). Breed affected dry matter intake and milk production and composition. Holstein cows had the highest dry matter intake (18.4±0.40kg of DM/d) and milk production (21.1±0.53kg of DM/d). Holstein and Jersey × Holstein cows had similar 4% fat corrected milk, fat yield, and protein yield; with the exception of fat yield, these were all higher than for Jersey cows. Milk fat concentration was highest for Jersey cows and lowest for Holstein cows, with the hybrid cows intermediate. Total FA and linolenic acid intake (1.09±0.023 and 0.58±0.012 kg/d, respectively) were highest for Holstein cows. In terms of milk FA, Holstein cows had higher contents of C14:1, cis-9 C18:1 and linoleic acid. In turn, Jersey and Jersey × Holstein cows had higher content of C16:0. Milk concentrations of neither the cis-9,trans-11 isomer of CLA nor its precursor, vaccenic acid, were affected by breed. Nevertheless, large variation between individual animals within breed grouping was observed for CLA and estimated Δ9-desaturase activity. There was some evidence for a negative heterotic effect on milk concentration of CLA, with the F1 hybrid cows having lower concentrations compared with the mid parent average. Plasma FA profile did not accurately reflect differences in milk FA composition. In conclusion, there was little evidence for either breed or beneficial heterotic effects on milk FA content with human health-promoting potential, though significant within-breed, interanimal variation was observed.

Incorporation profiles of isomers of conjugated linoleic acids (CLA) into raft and non‐raft lipids of CHO cells.

The aim of this study was to determine whether part of the biological effects of CLA is due to their incorporation into membrane lipids, and consequent modification of membrane raft function. The incorporation of four CLA isomers, trans 10‐cis 12 (A), trans 9‐trans 11 (B), cis 9‐trans 11 (C), and cis 9‐cis 11 (D), and of linoleic acid (LA) into raft lipids, and their positional distribution in phospholipids was studied. In all cases, more than 60% of the cellular CLA was present in the raft fraction. Part of the CLA was converted to the 18:3 and 20:2 derivatives by the cells. Of the total CLA incorporated, 60% was present in phosphatidylcholine (PC), and 30% in phosphatidylethanolamine (PE). Unlike LA, which was present almost exclusively in the sn‐2 position of phospholipids, 30–40% of A, C, and D, and up to 70% of B were found in the sn‐1 position of PC. About 65% of the total CLA in PE was present in sn‐1 position for all the isomers. In all cases endogenous 18:1 was the predominant fatty acid replaced by CLA, but saturated fatty acids were also replaced by B. Furthermore, A, C, and D inhibited the Δ9 desaturase activity, as evident from the decreased ratios of monunsaturates/saturates. CLA‐B, however, increased this ratio, indicating no inhibition of the enzyme by it. These results show that CLA incorporate preferably into raft lipids, with a large percentage in sn‐1 position of phospholipids, which may affect the membrane functions.

Whole blood fatty acid analysis with micromethod in cystic fibrosis and pulmonary disease

Objectives To assess fatty acid (FA) profiles in whole blood of 90 cystic fibrosis patients (CF) and 30 control subjects (C) and to correlate FA changes to the severity of respiratory disease. Methods Whole blood FA were assessed by GC with a micromethod-based analysis. Results Saturated and monounsaturated FA are higher, whereas polyunsaturated FA are lower in CF versus C with reduction of total n-6 FA, 22:5n-3 and 22:6n-3 (DHA). The product of linoleic acid (LA) × DHA, proposed as a marker for the disease, is 30% lower in CF than in C. Correlations with the severity of the respiratory disease are present for different FA and for the LA × DHA product. There is a reduction of Δ5 desaturase activity in CF, greater in severe disease, suggesting a basic metabolic alteration. Conclusions The micromethod-based analysis of blood FA facilitates the assessment of the FA status while confirming alterations of FA profiles already reported in specific blood compartments of CF.

Cloning, Tissue Expression Analysis, and Functional Characterization of Two Δ6-Desaturase Variants of Sea Bass (Dicentrarchus labrax L.)

Fish are the main source of the n-3 highly unsaturated fatty acids, which are crucial for human health. Their synthesis from C(18) precursors is mediated by desaturases and elongases, but the activity of these enzymes has not been conclusively established in marine fish species. This study reports the cloning, tissue expression, and functional characterization of a sea bass (Dicentrarchus labrax L.) Δ6-desaturase and one of its splicing variants. Two cDNAs with open reading frames of 1,346 and 1,354 bp were cloned and named D6D and D6D-V, respectively. Both deduced protein sequences (445 and 387 amino acids, respectively) contained two transmembrane regions and the N-terminal cytochrome b(5) domain with the HPGG motif characteristic of microsomal desaturases. D6D presents three histidine-rich regions, whereas in D6D-V, an insertion of eight nucleotides in the boundaries of exons 10 and 11 modified the third histidine-rich domain and led to insertion of a premature STOP codon, resulting in a shorter predicted protein. Quantitative real-time polymerase chain reaction assay of gene expression showed that D6D was highly expressed in the brain and intestine, and to a lesser extent, in muscle and liver; meanwhile, D6D-V was expressed in all tissues tested, but at level at least 200-fold lower than D6D. Functional analysis in yeast showed that sea bass D6D encodes a fully functional Δ6-desaturase with no residual Δ5-desaturase activity. This desaturase does not exhibit a clear preference for n-3 versus n-6 C(18) substrates. Interestingly, D6D-V is a nonfunctional protein, suggesting that the C-terminal end is indispensable for protein activity.

A Multiplexed Cell Assay in HepG2 Cells for the Identification of Delta-5, Delta-6, and Delta-9 Desaturase and Elongase Inhibitors

A multiplexed cell assay has been optimized to measure the activities of fatty acyl-CoA elongase, delta-5 desaturase (Delta5D), delta-6 desaturase (Delta6D), and delta-9 desaturase (Delta9D) together using (14)C-labeled tracers in HepG2 cells, which express the human stearoyl-CoA desaturase-1 isoform (SCD1) exclusively. The Delta5 and Delta9 desaturase activities are indexed by the efficient conversion of [1-(14)C]-eicosatrienoic acid (C20:3, cis-8,11,14) to (14)C-arachidonic acid (C20:4, cis-5,8,11,14) and the conversion of [1-(14)C]-stearic acid to (14)C-oleic acid (C18:1, cis-9), respectively. CP-74006 potently blocks the Delta5D activity with an IC(50) value of 20 nM and simplifies the metabolism of [1-(14)C]-alpha-linolenate (C18:3, cis-9,12,15) by accumulating (14)C-eicosatetraenoic acid (C20:4, cis-8,11,14,17) as the major (14)C-eicosatrienoic acid (C20:3, cis-11,14,17) and (14)C-docosatetraenoic acid (C22:4, cis-10,13,16,19) as the minor metabolites through Delta6 desaturation and elongation. This simplified metabolite spectrum enables the delineation of the Delta6D activity by comparing the combined Delta6D/elongase activity index of the (14)C-(C20:4/C18:3) ratio with the corresponding elongation index of the (14)C-(C20:3/C18:3) ratio following compound treatment. SC-26196 and sterculic acid specifically inhibit the Delta6D and Delta9D activities with an IC(50) value of 0.1 microM and 0.9 microM, respectively. This medium-throughput cell assay provides an efficient tool in the identification of specific desaturase and elongase inhibitors.

The Polyunsaturated Fatty Acid Composition of Hepatic and Plasma Lipids Differ by Both Sex and Dietary Fat Intake in Rats1–3

In rats and humans, females have higher liver and/or plasma docosahexaenoic acid (DHA) content than males. We hypothesized that the effect of variation in total fat or essential fatty acid intakes on liver and plasma fatty acid composition would differ between sexes. Rats were fed a low-fat soybean oil (LFS), high-fat soybean oil (HFS), or high-fat linseed oil (HFL) diet for 20 d. There were significant sex differences in LFS rats in proportions of (n-3) and (n-6) fatty acids in plasma and liver contingent on lipid class. Significant diet × sex interactions were observed for eicosapentaenoic acid (EPA), DHA, and arachidonic acid (AA) status. HFL females had a higher proportion of EPA in plasma and liver phosphatidylcholine (PC), DHA in liver triacylglycerol (TAG), and AA in plasma PC than HFS and LFS females. These findings show that the effect of varying dietary fat intake on (n-3) and (n-6) long-chain PUFA (LCPUFA) status is modified by sex. Liver phospholipid and TAG fatty acid product:substrate ratios suggested greater Δ6 desaturase (Δ6D) activity in females than in males. The HFL diet induced higher Δ6D mRNA expression compared with the LFS or HFS diets and HFL females had 10% higher expression of Δ6D mRNA than HFL males. Together, these findings show that sex is an important determinant of the effect of variations in fat and fatty acid intake on LCPUFA status, which may have implications for recommendations for fat and fatty acid intake in humans.

Role of hepatic desaturases in obesity-related metabolic disorders

This review aims to address the latest research on hepatic desaturases and metabolic disorders, with focus on stearoyl-CoA desaturase-1 (SCD-1) indices in observational studies. In animal studies, SCD-1 inhibition protects against features of the metabolic syndrome and is associated with improved hepatic insulin resistance and decreased steatosis. In human observational studies, higher estimated hepatic SCD-1 and Δ6-desaturase activities predict the metabolic syndrome, insulin resistance and mortality whereas Δ5-desaturase index is often inversely related. However, because the desaturase activities in the liver and adipose tissue may not be regulated in parallel, it is important to define used lipid fractions when comparing studies. It is also important to take the background diets of the populations into account when comparing studies. Moreover, there may be a divergence in desaturase regulation depending on glycaemic control among individuals. Increased SCD-1 indices reflecting liver desaturase activity have been associated with insulin resistance, fatty liver, the metabolic syndrome and mortality. However, it remains to be determined if high hepatic SCD-1 activity plays a direct role in the development of metabolic disorders or rather is a marker for an unfavourable diet or hepatic insulin resistance.

Fatty acid composition of milk from multiparous Holstein cows treated with bovine somatotropin and fed n-3 fatty acids in early lactation

Multiparous cows (n=59) were blocked by expected calving date and previous milk yield and assigned randomly to treatments to determine effects of bovine somatotropin (bST; Posilac, Monsanto Animal Agricultural Group, St. Louis, MO) and source of dietary fat on milk fatty acid composition during the first 140 d in milk. Diets were provided from calving and included whole, high-oil sunflower seeds (SS; 10% of dietary dry matter; n-6/n-3 ratio of 4.6) as a source of linoleic acid or a mixture of Alifet-High Energy and Alifet-Repro (AF; Alifet USA, Cincinnati, OH; 3.5 and 1.5% of dietary dry matter, respectively; n-6/n-3 ratio of 2.6) as a source of protected n-3 fatty acids (15.7% 18:3, 1.3% 20:5, and 1.3% 22:6). Treatments were derived from a 2×2 combination of supplemental fat source (SS, AF) and with 0 (SSN, AFN) or 500 (SSY, AFY) mg of bST administered every 10 d from 12 to 70 d in milk and at 14-d intervals thereafter. Milk fatty acid composition was determined in samples collected from 32 cows (8 complete blocks) during wk 2, 8, and 20 of lactation. Data were analyzed as repeated measures using mixed model procedures to determine the effects of diet, bST, week of lactation, and their interactions. Proportions of 18:3 (4.02 vs. 3.59±0.16%), 20:5 (0.52 vs. 0.41±0.02%), and 22:6 (0.11 vs. 0.02±0.02%) were greater and the n-6/n-3 fatty acid ratio (7.40 vs. 8.80±0.30) was reduced in milk from cows fed AF compared with SS. Proportions of de novo-synthesized fatty acids increased and preformed fatty acids decreased as lactation progressed, but bST administration delayed this shift in origin of milk fatty acids. Transfer efficiency of 18:3, 20:5, and 22:6 from AF to milk fat averaged 36.2, 4.9, and 5.2%, respectively. These efficiencies increased as lactation progressed, but were delayed by bST. Apparent mammary Δ9-desaturase activity and milk conjugated linoleic acid (cis-9, trans-11 conjugated linoleic acid) content increased through the first 8 wk of lactation. Based on the product-to-substrate ratio of 14:1/14:0 fatty acids in milk, there was an interaction of diet and bST because bST decreased apparent Δ9-desaturase activity in SSY cows but increased it in AFY cows (0.10, 0.09, 0.08, and 0.09±0.01 for SSN, SSY, AFN, and AFY, respectively). Feeding Alifet-Repro increased n-3 fatty acids in milk and bST prolonged the partitioning of dietary fatty acids into milk fat.

Effect of Cyclosporine and Sirolimus on Fatty Acid Desaturase Activities in Cultured HEPG 2 Cells

The aim of the present work was to evaluate the influence of cyclosporine (CsA) and sirolimus (SRL) on fatty acid (FA) desaturase activities. These enzymes (named Δ9, Δ6, and Δ5 desaturases) catalyze reactions leading to the biosynthesis of n-9, n-6, and n-3 FA families. n-3 FA family, derived from alpha-linolenic acid, is involved in the prevention of vascular events, which appear after successful kidney transplantation. Five groups of HepG 2 cells in culture were treated with either CsA (1 μg/μL and 2 μg/μL) or SRL (10 ng/mL and 20 ng/mL) for 3 days, including a control group without immunosuppressive treatment. We studied the incorporation and metabolic conversion of radioactive [1- 14C]palmitic, linoleic, and eicosatrienoic acids. We also analyzed fatty acid composition. The distribution of radioactive metabolic products after incubation of these cells with [1- 14C]palmitic acid revealed a decrease in Δ9 desaturase activity in the presence of each immunosuppressive drug: CsA = 0.61 ± 0.01; SRL = 0.59 ± 0.04 versus control = 0.79 ± 0.05 ( P < .01). We observed a significant increase in Δ6 and Δ5 desaturase activities under the influence of the immunosuppressive drugs: radiolabeled linoleic acid (CsA: 0.93 ± 0.04; SRL: 1.02 ± 0.03 vs control 0.60 ± 0.03; P < .01) and eicosatrienoic acid (CsA: 1.12 ± 0.02; SRL: 1.07 ± 0.01 vs control 0.75 ± 0.01; P < .01). In conclusion, CsA and SRL modulated the biosynthesis of polyunsaturated FAs, decreasing Δ9 desaturase and increasing Δ6 and Δ5 desaturase activities.