Gamma-linolenic acid provides additional protection against ventricular fibrillation in aged rats fed linoleic acid rich diets

Ligation of the coronary artery in rats produces severe ventricular fibrillation (VF) and malignant cardiac arrhythmia. Mortality increases with the age of the animal. Diets rich in saturated fatty acids (SF) but low in linoleic acid (LA) increase, but diets high in LA and low in SF decrease the severity of VF and mortality in older animals. The effects of an LA enriched diet can be blocked by inhibition of cyclooxygenase suggesting that conversion of LA to eicosanoids is central to the development of VF. Conversion of LA to gamma-linolenic acid (GLA) via delta-6 desaturase is the first step in the process. The activity of delta-6 desaturase declines with age. Thus inclusion of GLA in the diet of older animals may provide an additional benefit over LA alone. Dietary supplements of evening primrose oil (EPO) to one year old rats reduced ischaemic VF more than a supplement of sunflower seed oil (SSO) without GLA. Substitution of borage oil (more GLA than EPO but less LA than either EPO or SSO) was without additional benefit.     

Enhanced prostaglandin synthesis as a mechanism for inhibition of melanoma cell growth by ascorbic acid

Both ascorbic acid and the 1-series prostaglandins have been reported to be important regulators of cell growth and since ascorbic acid also increases the synthesis of the 1-series prostaglandins, it is possible that the effects of ascorbic acid on cell growth might be mediated by changes in 1-series prostaglandin synthesis induced by ascorbic acid. This study attempted to examine this possible relationship. The effects of ascorbic acid, prostaglandin E1 and the essential fatty acid precursors of the prostaglandins, linoleic acid and gamma-linolenic acid on the in vitro growth of transformed BL6 murine melanoma cells and untransformed monkey kidney (LLCMK) cells was determined. The effects of ascorbic acid addition on the growth inhibitory effect of the essential fatty acids and on the activity of delta-6-desaturase, a key enzyme in 1-series prostaglandin synthesis were also examined. Addition of ascorbic acid, prostaglandin E1 and both essential fatty acids was found to reduce BL6 growth while PGE1 and to a lesser extent the essential fatty acids reduced LLCMK cell growth. The growth inhibitory effect of the essential fatty acids was enhanced by ascorbic acid which was also found to stimulate delta-6-desaturase activity in BL6 cells. The growth inhibitory effect of ascorbic acid on BL6 cells may thus be mediated by changes in prostaglandin synthesis through an association with the metabolism of the essential fatty acid precursors of the prostaglandins.     

The effects of evening primrose oil, safflower oil and paraffin on plasma fatty acid levels in humans: Choice of an appropriate placebo for clinical studies on primrose oil

In a number of diseases, plasma levels of linoleic acid are normal or elevated while those of gamma-linolenic acid (18:3n-6, GLA) and further metabolites are below normal. Evening primrose oil (EPO), similar to safflower oil (SFO) except that it contains 8-9% of 18:3n-6, has been proposed as a therapeutic agent in these diseases, such as atopic eczema. There is argument as to whether an appropriate placebo for clinical studies on EPO should be an inert material such as paraffin, or a linoleic acid--containing oil such as SFO. We have therefore compared in normal humans the effects on plasma fatty acids of administering EPO, SFO and paraffin for 10 days. Paraffin had no effect on any fatty acid in any fraction. EPO raised the level of 20:3n-6 (dihomo-gamma-linolenic acid, DGLA) the immediate metabolite of GLA but had no significant effect on arachidonic acid. In surprising contrast, SFO raised the levels of linoleic and of arachidonic acids, without raising those of DGLA. This suggests that linoleic acid may be rapidly converted to arachidonic acid by a tightly linked enzyme sequence: GLA, in contrast, may be rapidly converted to DGLA but then only slowly on to arachidonic acid. These results are consistent with recent in vitro observations by others on rat hepatocytes and human fibroblasts.     

Delta-6 desaturase from borage converts linoleic acid to gamma-linolenic acid in HEK293 cells

Gamma-linolenic acid (GLA, 18:3 n6) is an essential polyunsaturated fatty acid of the omega-6 family and is found to be effective in prevention and/or treatment of various health problems. In this study, we evaluated the possibility of increasing γ-linolenic acid contents in mammalian cells using the delta-6 gene from Borago officinalis. The borage Δ6-desaturase gene (sDelta-6) was codon-optimized and introduced into HEK293 cells by lipofectin transfection. Co-expression of GFP with sDelta-6 and RT-PCR analysis indicated that sDelta-6 could be expressed in mammalian cells. Subsequently, the heterologous expression of borage Δ6-desaturase was evaluated by fatty acid analysis. Total cellular lipid analysis of transformed cells fed with linoleic acid (LA 18:2 n6) as a substrate showed that the expression of sDelta-6 resulted in an 228–483% (p < 0.05) increase of GLA when compared with that in the control cells. The highest conversion efficiency of LA into GLA in sDelta-6⁺ cells was 6.9 times higher than that in the control group (11.59% vs. 1.69%; p < 0.05). Our present work demonstrated that the sDelta-6 gene from borage could be functionally expressed in mammalian cells, and could convert LA into GLA. Furthermore, this study may pave the way to generate transgenic livestock that can synthesise GLA.     

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.     

Beneficial effects of gamma linolenic acid supplementation on nerve conduction velocity, Na+, K+ ATPase activity, and membrane fatty acid composition in sciatic nerve of diabetic rats

Metabolic and vascular abnormalities are implicated in the pathogenesis of diabetic neuropathy. Two principal metabolic defects are altered lipid metabolism resulting from the impairment of delta-6-desaturase, which converts linoleic acid (LA) into gamma linolenic acid (GLA), and reduced nerve Na+, K+ ATPase activity. This reduction may be caused by a lack of incorporation of (n-6) fatty acids in membrane phospholipids. Because this ubiquitous enzyme maintains the membrane electrical potential and allows repolarization, disturbances in its activity can alter the process of nerve conduction velocity (NCV). We studied the effects of supplementation with GLA (260 mg per day) on NCV, fatty acid phospholipid composition, and Na+, K+ ATPase activity in streptozotocin-diabetic rats. Six groups of 10 rats were studied. Two groups served as controls supplemented with GLA or sunflower oil (GLA free). Two groups with different durations of diabetes were studied: 6 weeks with no supplementation and 12 weeks supplemented with sunflower oil. To test the ability of GLA to prevent or reverse the effects of diabetes, two groups of diabetic rats were supplemented with GLA, one group for 12 weeks and one group for 6 weeks, starting 6 weeks after diabetes induction. Diabetes resulted in a 25% decrease in NCV (P < 0.0001), a 45% decrease in Na+, K+ ATPase activity (P < 0.0001), and an abnormal phospholipid fatty acid composition. GLA restored NCV both in the prevention and reversal studies and partially restored Na+, K+ ATPase activity in the preventive treatment group (P < 0.0001). These effects were accompanied by a modification of phospholipid fatty acid composition in nerve membranes. Overall, the results suggest that membrane fatty acid composition plays a direct role in NCV and confirm the beneficial effect of GLA supplementation in diabetic neuropathy.     

Effect of L-arginine-nitric oxide system on the metabolism of essential fatty acids in chemical-induced diabetes mellitus

Several studies have shown that the activities of delta-6-desaturase and delta-5-desaturase are depressed in experimental diabetes and in humans with insulin- and non-insulin-dependent diabetes mellitus (type I and type II diabetes mellitus respectively). Furthermore, treatment with insulin is known to correct the defects in desaturases in rats and humans with diabetes, especially in type I. In a recent study, we demonstrated that L-arginine and nitric oxide can prevent alloxan-induced beta cell damage, and the severity of diabetes, and restore the antioxidant status to near normal levels. But, no information is available as to the relationship between L-arginine-nitric oxide system and the metabolism of essential fatty acids in diabetes mellitus. In the present study, it was noted that the plasma levels of saturated fatty acids: stearic and palmitic were increased where as unsaturated fatty acids such as oleic, linoleic, gamma-linolenic and eicosapentaenoic acids (OA, LA, GLA and EPA respectively) were decreased in alloxan-induced diabetic rats. In the liver phospholipid (PL) fraction, GLA, DGLA (dihomo-GLA) and alpha-linolenic acid (ALA) were decreased in the alloxan-treated group; in the muscle PL fraction, LA, GLA and DGLA were low, whereas an increase in the saturated fatty acid content was noted. L-arginine (the precursor of nitric oxide) and sodium nitroprusside (a nitric oxide donor) treatment of alloxan-induced diabetic rats enhanced the levels of LA, GLA and DGLA. Further, nitric oxide synthase inhibitor, L-NMMA, prevented this beneficial action of L-arginine-nitric oxide system on essential fatty acid metabolism. The abnormalities in the essential fatty acid metabolism observed also reverted to normalcy following control of diabetes with insulin. These results indicate that nitric oxide can enhance the activities of delta-6- and delta-5 desaturases, which are depressed in diabetes, and suggests that there is a close interaction between L-arginine-nitric oxide system and the metabolism of essential fatty acids.     

Effects of Triterpenes on the Mutagenicities of Various Mutagens toward Salmonella

The effects of different dietary levels of gamma-linolenic acid (GLA) on lipid metabolism was studied in rats using a combination of evening primrose oil (EPO) and palm oil (PLO). EPO compared to PLO significantly reduced liver cholesterol and triglyceride after 4 weeks of feeding, and the effect remained even when EPO was mixed with PLO at the same weight basis. The serum triglyceride level also tended to be low on feeding EPO. Neither liver Δ6-desaturase and phospholipase A₂ activities nor aortic production of prostacyclin and thromboxane A₂ production by platelets were influenced significantly by the fat type, suggesting a peculiar effect of PLO. The percentage of arachidonic acid in liver, serum, and aortic phosphatidylcholine depended on the dietary level of GLA. A more distinct increase in arachidonic acid was observed in tissue triglycerides of rats fed EPO. GLA appears to exert favorable effects on lipid metabolism even when the P/S ratio was lowered from 13.7 of EPO to 1.8 of the 1:1 mixture of EPO and PLO.     

Effects of Various Polyunsaturated Fatty Acids on Blood Cholesterol and Eicosanoids in Rats

The hypocholesterolemic efficacy of various polyunsaturated fatty acids was compared in rats given cholesterol-enriched diets with (0.004%) or without indomethacin, the cyclooxygenase inhibitor. Evening primrose oil (EPO, linoleic+ γ-linolenic), safflower oil (SFO, linoleic) or soybean oil (SBO, linoleic + α-linolenic) were added to diets at the 10% level. The serum cholesterol level of the EPO group was consistently lower than the other groups and after 3 weeks, it was significantly different from the SFO group without indomethacin and the SBO group with indomethacin. In rats fed EPO, the aorta tended to produce more prostacyclin whereas the concentration of plasma thromboxane B₂ was much lower than in rats fed SFO or SBO. The effects of indomethacin on these eicosanoids were less evident in rats fed EPO. Thus, in addition to the hypocholesterolemic action of β-linolenic acid (GLA) in preference to linoleic and possibly α-linolenic acid, GLA appears to cultivate an environment suitable for the prevention of carbiovascular disease even in the presence of excess cholesterol in the diet.     

Kinetic analysis of delta-6-desaturation in liver microsomes: influence of gamma-linoleic acid dietary supplementation to young and old rats.

Previous experiments demonstrated the ability-of a gamma-linoleic acid (GLA) dietary supplementation (as evening primrose oil--EPO) to counteract the fall off in delta-6-desaturase (D6D) activity of linoleic acid and alpha-linoleic acid in aged rats. Kinetic parameters of the D6D were determined in order to test the possibility that there may be a significant influence of GLA administration to young and aged rats on the Vm and Km values for 6-desaturation of both the substrates. In young rats GLA supplementation did not affect the kinetic parameters, while in old rats it produced an increase of Vm values of 6-desaturation for both the substrates. Thus the administration of small doses of GLA to old rats might offer substantial protection against the loss of D6D affinity observed in aging, enhancing the capacity of the enzyme itself.     

Dose-dependent effects of dietary gamma-linolenic acid on rat spleen lymphocyte functions.

Feeding rodents a diet rich in evening primrose oil (EPO), which contains 5-10 g gamma-linolenic acid (GLA)/100 g total fatty acids, has been shown to decrease lymphocyte proliferation and natural killer cell activity. However, EPO contains a very high level of linoleic acid which itself can affect lymphocyte functions and it is not clear to what extent the effects of EPO can be attributed to GLA. The current study investigated the effect of two levels of GLA in the rat diet upon immune cell functions; the level of linoleic acid was maintained below 30 g/100 g total fatty acids. Weanling rats were fed on high fat (178 g/kg) diets which contained 4.4 g or 10 g GLA/100 g total fatty acids in place of a proportion of linoleic acid. The total polyunsaturated fatty acid content and the n-6 to n-3 polyunsaturated fatty acid ratio of the diet were maintained at 35 g/100 g total fatty acids and 7, respectively. The fatty acid compositions of the serum and of spleen leukocytes were markedly influenced by that of the diet, with an increase in the proportions of GLA and dihomo-gamma-linolenic acid when the diets containing GLA were fed; these diets also increased the proportion of arachidonic acid in spleen leukocytes. Spleen lymphocyte proliferation in response to concanavalin A was significantly reduced (by 60%) by feeding the diet containing the higher level of GLA, but not by the diet containing the lower level of GLA. Spleen natural killer cell activity and prostaglandin E (PGE) production by spleen leukocytes were not significantly affected by inclusion of GLA in the diet, although there was a tendency towards decreased natural killer cell activity by cells from rats fed the high GLA diet. Thus, this study shows that dietary GLA is capable of altering the fatty acid composition of cells of the immune system and of exerting some immunomodulatory effects, but that the level of GLA in the diet must exceed 4.4 g/100 g total fatty acids for these effects to become apparent.     

The effect of chemical hepatocarcinogenesis on liver phospholipid composition in rats fed N-6 and N-3 fatty acid-supplemented diets.

The effect of dietary fats on essential fatty acid metabolism in rats subjected to chemically induced hepatocarcinogenesis was studied. Sixty male rats were fed a diet supplemented with one of the following three oil compositions: 10% hydrogenated coconut oil (HCO); 5% hydrogenated coconut oil and 5% gamma-linolenic acid (18:3n-6)-rich evening primrose oil (EPO); or 5% hydrogenated coconut oil and 5% marine oil (FO). Half of the animals in each dietary regimen were subjected to hepatocarcinogenesis induction using diethylnitrosamine and 2-acetylaminofluorene (2-AAF) followed by partial hepatectomy, whereas the other half underwent hepatectomy without receiving diethylnitrosamine and 2-acetylaminofluorene. Liver phospholipid composition was analyzed. In comparison to the HCO group, the EPO group showed raised levels of arachidonic acid (20:4n-6) and suppressed n-3 fatty acids. The FO group, on the other hand, showed suppressed levels of n-6 and increased n-3 fatty acids. Hepatocarcinogenesis suppressed the level of 20:4n-6 and this effect was greater in the FO rats. The levels of dihomo-gamma-linolenic acid (20:3n-6) were increased by the hepatocarcinogenic treatment, and this effect was further accentuated in the EPO rats. These results suggest that hepatocarcinogenesis may suppress the activity of delta-5-desaturase, which may be one of the reasons why tumor cell membranes have low levels of long chain fatty acids, especially 20:4n-6 cells, and have an impaired capacity to undergo lipid peroxidation.     

Structural characterization of triacylglycerol in several oils containing gamma-linolenic acid

The differences are reported in the triacylglycerol (TG) structures of oils containing gamma-linolenic acid (GLA) from Oenothera biennis Linn seed oil (OBLO) from the wild plant, evening primrose seed oil (EPO) from a cultured plant, and bio-GLA oil (BIO) from a mold, the physiological functions of which were ascertained by animal testing. Reverse-phase high-performance liquid chromatographic separation detected 12 TG peaks each for OBLO and EPO, and 28 TG peaks for BIO. TG-containing GLA were composed of five molecular species each in OBLO and EPO, and ten molecular species in BIO. The totals of the molecular species containing GLA were 29.8% in OBLO, 23.8% in EPO, and 56.6% in BIO. In OBLO, the GLA level at the sn-2 position of the major TG species was higher than that in EPO. In BIO, the GLA level at the sn-2 position of the major TG species was lower than those in OBLO and EPO.     

Lipid formation and γ-linolenic acid production by Mucorales fungi grown on sunflower oil

Lipid formation and γ-linolenic acid (GLA) production by 48 species of Mucorales fungi grown on sunflower oil (which consists of 70% linoleic acid ; LA) were studied. The strains accumulated 42·7–65·8% lipid in biomass (7·66–13·39 g l⁻¹). Eight cultures produced more than 200 mg GLA l⁻¹. Highest GLA yields exhibited Mucor mucedo CCF-1384 and Cunninghamella echinulata CCF-103 (379 and 373 mg l⁻¹, respectively). Mortierella alpina CCF-185 synthesized 465 mg l⁻¹ arachidonic acid. While the decrease of LA utilization index (ratio of LA content of cell lipid/LA content of oil source) was accompanied with growth of delipidized biomass and with reduction of lipid accumulation within the cells, high lipid yield was as a consequence of the direct oil source incorporation into intracellular lipid.     

The effects of hydrogenated coconut oil, safflower oil, and evening primrose oil on development of hypertension and sodium handling in spontaneously hypertensive rats

Spontaneously hypertensive rats (SHR) were fed a basal regular diet (BD) or three different fat-supplemented diets which contained 10% hydrogenated coconut oil (HCO), 10% safflower oil (SFO), or 10% evening primrose oil (EPO). The rats received these four different diets from 4 weeks to over 24 weeks of age. The development of hypertension in SHR was significantly retarded in the EPO-supplemented animals. The blood pressure was lower in the SFO group animals as compared with the BD and HCO groups, but this did not reach significance. Sodium excretion rate in young SHR was increased in the EPO group compared with the HCO and SFO groups, and the urinary K/Na ratio was decreased in the EPO group compared with the HCO and EPO groups. Water intake and urine volume were increased in the SFO group as compared with the HCO and EPO groups. Sodium concentration in erythrocytes was decreased in the rats receiving SFO. Pressor responses to norepinephrine and angiotensin II were enhanced in the EPO and SFO groups as compared with the basal chow group. These data suggest that a dietary supplementation of EPO which contains a substantial amount of gamma-linolenic acid consistently lowers blood pressure in SHR. The mechanism is uncertain, but the effects on sodium handling may in part be responsible for the retardation of the development of hypertension. There was a difference between the EPO and the SFO groups in sodium--water handling, and to some extent in the blood pressure development in SHR.     

Fermentation process development of recombinant Hansenula polymorpha for gamma-linolenic acid production

Development in the strain and the fermentation process of Hansenula polymorpha was implemented for the production of gamma-linolenic acid (GLA, C18:3 delta 6,9,12), which is an n-6 polyunsaturated fatty acid (PUFA) and has been reported to possess a number of health benefits. The mutated delta 6-desaturase (S213A) gene of Mucor rouxii was expressed in H. polymorpha under the control of the methanol oxidase (MOX) promoter. Without utilization of methanol a high cell-density culture of the yeast recombinant carrying the delta 6-desaturase gene was achieved by fed-batch fermentation using glycerol-limited conditions. The delta 6-desaturated products, octadecadienoic acid (C18:2 delta6,9), GLA and stearidonic acid (C18:4 delta6,9,12,15), accumulated at high levels under the derepression condition. The GLA production was also optimized by adjusting specific growth rates. The results show that the specific growth rate affected both lipid content and fatty acid composition of the GLA-producing recombinant. Among the various specific growth rates studied, the highest GLA concentration, which was at of 697 mg/l, was obtained in the culture with the specific growth rate of 0.08 /h. Interestingly, the fatty acid profile of the yeast recombinant bearing the Mucor delta 6-desaturase gene was similar to that of blackcurrant oil with both containing similar proportions of n-3 and n-6 essential fatty acids.     

Delta-6-desaturase and delta-5-desaturase in human Hep G2 cells are both fatty acid interconversion rate limiting and are upregulated under essential fatty acid deficient conditions

Essential fatty acids are interconverted by desaturases and elongases to eicosanoid precursors. In essential fatty acid deficiency (EFAD) an increased hepatic interconversion of linoleic acid (18:2) to arachidonic acid (20:4n-6) has been demonstrated in vivo. We now cultured Hep G2 cells under EFAD conditions. 20:3n-6 appeared in EFAD cells, but also in controls. After adding ¹⁴C-18:2 to the medium, interconversion products and their distribution in different lipids were studied by HPLC. When trace amounts 18:2 were incubated, 38% were converted by the EFAD cells after 21 h, vs 6% by controls. 20% was converted to 20:4 by EFAD cells vs 14% by controls. EFAD cells preferentially distributed more 18:2 and conversion products to neutral fats and to phosphatidyl ethanolamine, but less to cardiolipin than controls did, when incubated with trace amount 18:2, but not with 1 mM 18:2. A relative accumulation of radioactivty in 20:3 was observed. In conclusion; in EFAD Hep G2 cells delta-6- and delta-5-desaturase both were found to be upregulated and eicosanoid precursors were distributed more into phosphatidyl ethanolamine. Delta-5-desaturase had a rate limiting property as well as delta-6-desaturase.     

Fatty acid changes in liver choline and ethanolamine glycerophospholipids in aspirin-treated rats fed linoleate, gamma-linolenate and fish oil

The effects of dietary linoleic acid, gamma-linolenic acid and marine fatty acids on the development of aspirin-induced gastric hemorrhage and the distribution of liver glycerophospholipid fatty acids in fat-deficient growing rats were studied. Aspirin (100 mg/day)-treated and nontreated rats were fed for 7 days, a mixed diet of 2.5% safflower oil and 7.5% hydrogenated coconut oil (SFO/HCO) or 7.5% fish oil (SFO/FO), or 2.5% gamma-linolenate concentrate and 7.5% fish oil (GLA/FO). Gastric hemorrhage was induced in animals by aspirin treatment to various extents. It was not affected by FO feeding, but was significantly alleviated by GLA feeding. Aspirin treatment reduced the proportions of 20:4n-6 in liver phosphatidylcholine. FO feeding (in SFO/FO and GLA/FO rats) further reduced the 20:4n-6 level and replaced it by n-3 fatty acids. GLA feeding, on the other hand, elevated the proportion of 20:4n-6. As a result, the reduction of 20:4n-6 by fish oil feeding, was less significant in GLA/FO rats than in SFO/FO rats. The degree of gastric hemorrhage appeared to relate negatively to the levels of 20:4n-6 in liver phosphatidylcholine, and to the sum of 20:4n-6 and 20:5n-3 when FO was included in the diet. It is suggested that long-chain polyunsaturated fatty acids (20:4n-6 and 20:5n-3) per se in addition to being precursors of prostaglandins, may also affect the development of gastric hemorrhage, possibly by modulating the permeability of cell membranes in the gastric mucosa.     

Sex differences in n-3 and n-6 fatty acid metabolism in EFA-depleted rats

We studied the effect of sex on the distribution of long-chain n-3 and n-6 fatty acids in essential fatty acid-deficient rats fed gamma-linolenate (GLA) concentrate and/or eicosapentaenoate and docosahexaenoate-rich fish oil (FO). Male and female weanling rats were rendered essential fatty acid deficient by maintaining them on a fat-free semisynthetic diet for 8 weeks. Thereafter, animals of each sex were separated into three groups (n = 6) and given, for 2 consecutive days by gastric intubation, 4 g/kg body wt per day of GLA concentrate (containing 84% 18:2n-6), n-3 fatty acid-rich FO (containing 18% 20:5n-3 and 52% 22:6n-3), or an equal mixture of the two oil preparations (GLA + FO). The fatty acid distributions in plasma and liver lipids were then examined. GLA treatment increased the levels of C-20 and C-22 n-6 fatty acids in all lipid fractions indicating that GLA was rapidly metabolized. However, the increases in 20:3n-6 were less in females than those in males, while those in 20:4n-6 were greater, suggesting that the conversion of 20:3n-6 to 20:4n-6 was more active in female than in male rats. FO treatment increased the levels of 20:5n-3 and 22:6n-3 and reduced those of 20:4n-6. The increase in n-3 fatty acids was greater in females than that in males and the reduction in 20:4n-6 was smaller. Consequently, the sum of total long-chain EFAs incorporated was greater in females than that in males. The administration of n-3 fatty acids also reduced the ratio of 20:4n-6 to 20:3n-6 in GLA + FO-treated rats indicating that n-3 fatty acids inhibited the activity of delta-5-desaturase. However, this effect was not affected by the sex difference.     

Molecular cloning and expression analysis of a delta 6-fatty acid desaturase gene from Rhizopus stolonifer strain YF6 which can accumulate high levels of gamma-linolenic acid

The delta 6-desaturase gene was cloned from Rhizopus stolonifer, which could accumulate up to 49% of gamma-linolenic acid (GLA, C18:3 Δ^6,9,12) to the total fatty acids. The cloned DNA contains a 1,380 bp open reading frame encoding a protein of 460 amino acids, which showed high similarity to those of fungal delta 6-desaturases with three conserved histidine-rich motifs and HPGG motif. Notably, this deduced sequence had a shorter C-terminus. Results demonstrated that the cDNA sequence exhibited delta 6-desaturase activity by accumulation of about 22.4 % of GLA to the total fatty acids in the recombinant Pichia pastoris strain GS115.