Effects of dietarytrans acids on the biosynthesis of arachidonic acid in rat liver microsomes

Effects of dietarytrans acids on the interconversion of linoleic acid was studied using the liver microsomal fraction of rats fed a semipurified diet containing fat supplements of safflower oil (SAFF), hydrogenated coconut oil (HCO) at 5 and 20% levels or a 5% level of a supplement containing 50.3% linolelaidic and 24.3% elaidic acids devoid ofcis,cis-linoleic acid (TRANS). Growth rate was suppressed to a greater extent with the animals fed the 20% than the 5% level of the HCO-supplemented diets and still further by the TRANS diet compared to the groups fed the SAFF diets. Food intake was greater in the groups fed the HCO than the SAFF-supplemented diets, demonstrating the marked effect of an essential fatty acid (EFA) deficiency on feed efficiency. In contrast to an EFA deficiency produced by the HCO supplement, which stimulated the in vitro liver microsomal biosynthesis of arachidonic acid, diets containing the TRANS supplement exacerabated the EFA deficiency and depressed 6-desaturase activity of the liver microsomal fraction. The liver microsomal fraction of the animals receiving this supplement also was more sensitive to fatty acid inhibition of the desaturation of linoleic acid than those obtained from animals fed either the SAFF or HCO diets. It is suggested that dietarytrans acids alter the physical properties of the 6-desaturase enzyme system, suppressing its activity, which increases the saturation of the tissue lipids and, in turn, the requirement for EFA or polyunsaturated fatty acids.     

The in vivo incorporation of labeled linoleic acid, α-linolenic acid and arachidonic acid into rat liver lipids

The incorporation of 1-¹⁴C-linoleic acid, 1-¹⁴C-α-linolenic acid and 1-¹⁴C-arachidonic acid into rat liver lipids was measured and the per cent distribution of radioactivity into the different lipid fractions determined. Normal rats were injected into the portal vein with the labeled solutions during a one minute period. Livers were quickly frozen, pulverized, and the lipids extracted and fractioned by thin layer chromatography. No significant differences were observed in the amounts of labeled fatty acids incorporated per gram of rat liver. While 1-¹⁴C-linoleic acid and 1-¹⁴C-α-linolenic acid were found in appreciable amounts in the 1,2 diacylglycerol fraction, about one fifth as much 1-¹⁴C-arachidonic acid was esterified in this fraction. 1-¹⁴C-arachidonic acid was the leading acid esterified in the phospholipid fractions.     

Dietary cod liver oil decreases arachidonic acid in rat gastric mucosa and increases stress-induced gastric erosions

The purpose of this study was to examine the influence of long-term feeding of dietary fat rich in either n−3 or n−6 fatty acids on the availability of arachidonic acid (20∶4n−6) in major phospholipids of gastric mucosa in rats. Three groups of male Wistar rats were fed either a standard diet, a cod liver oil-enriched diet (10% by weight), or a corn oil-enriched diet (10% by weight) for 8 mon. Dietary cod liver oil significantly reduced the level of 20∶4n−6 in phosphatidylcholine (PC) and in phosphatidylethanolamine (PE) of gastric mucosa. The loss of 20∶4n−6 was compensated for by eicosapentaenoic acid (20∶5n−3) in PC, whereas the decrease in 20∶4n−6 in PE corresponded to the increase in three n−3 fatty acids: 20∶5n−3, docosapentaenoic acid (22∶5n−3), and docosahexaenoic acid (22∶6n−3). The level of 20∶5n−3 was higher than the level of 22∶6n−3 both in PC and PE of mucosa in rats fed cod liver oil. Diets supplemented with corn oil increased the level of 18∶2n−6 but decreased the monoene fatty acids 16∶1 and 18∶1n−7 in PC but not in PE of gastric mucosa. The 20∶4n−6 levels of both PC and PE were markedly reduced by dietary cod liver oil, to about one-third of control levels. Similar changes were also observed in the stomach wall. Gastric erosions were observed in all rats exposed to restriction stress, but this form of stress induced twice the number of erosions in rats fed fish oil compared to control rats or rats fed corn oil. We conclude that a diet rich in fish oil altered the balance between n−6 and n−3 fatty acids in major gastric mucosal phospholipids, markedly reduced the availability of 20∶4n−6, and increased the incidence of gastric erosions induced by restriction or emotional stress.     

Regulation of the metabolism of linoleic acid to arachidonic acid in rat hepatocytes

When 5×10⁶ hepatocytes were incubated for 40 min with from 0.15 to 0.60 mM [1-¹⁴C]linoleic acid, [1-¹⁴C]6,9,12-octadecatrienoic acid, or [1-¹⁴C]8,11,14-eicosatrienoic acid, there was a concentration-dependent acylation of radioactive metabolites into both triglycerides and phospholipids. When the concentration of either [1-¹⁴C]linoleic acid or [1-¹⁴C]8,11,14-eicosatrienoic acid exceeded 0.3 mM, there was no further increase in the metabolism of either fatty acid to other (n−6) metabolites. When the concentration of [1-¹⁴C]6,9,12-octadecatrienoic acid exceeded 0.15 mM, there was an apparent substrate-induced inhibition in its metabolism to 8,11,14-eicosatrienoic acid. With all three substrates (0.3 mM), there was time-dependent metabolism to other (n−6) acids. Cells then were incubated simultaneously with 0.3 mM [1-¹⁴C]linoleic acid along with 0.15 to 0.45 mM 6,9,12-octadecatrienoic acid or 8,11,14-eicosatrienoic acid. These exogenous nonradioactive (n−6) acids suppressed but did not abolish the conversion of [1-¹⁴C]linoleate to radioactive arachidonate. These findings suggest that some linoleate is converted to arachidonate without intracellular mixing of 6,8,12-octadecatrienoic or 8,11,14-eicosatrienoic acids. This hypothesis is supported by the finding that exogenous linoleate did not markedly affect the metabolism of [1-¹⁴C]6,9,12-octadecatrienoic or [1-¹⁴C]8,11,14-eicosatrienoic acid by microsomal chain elongating or desaturating enzymes.     

Effect of dietary fatty acids on Δ5 desaturase activity and biosynthesis of arachidonic acid in rat liver microsomes

The effect of different fatty acids supplemented to a fat-free diet on the activity of Δ5 desaturase was studied. Fat-free diet produces a reduction in the conversion of eicosa-8,11,14-trienoic acid to arachidonic acid. The addition of thecis-ω6 acids, linoleic, γ-linolenic or arachidonic to the diet produces an increase of eicosatrienoic acid desaturation, shifting Δ5 desaturase activity towards the controls on a balanced diet. This reactivation is apparently produced by induction of enzyme biosynthesis since linoleate effect was suppressed by simultaneous cycloheximide injection. On the contrary, no changes in Δ5 desaturation activity were found when the diet was supplemented with palmitic or 9-trans,12-trans-linoleic acid. The changes on the activity of Δ5 desaturase were compared with the fatty acid composition of plasma and liver microsomes.     

Changes in liver fatty acid unsaturation after partial hepatectomy in the rat

The purpose of this study was to assess changes in the degree of fatty acid unsaturation in rat liver after partial hepatectomy. This is the first study in which liver fatty acid unsaturation has been analyzed over a long period of regeneration until day 28 after operation. The relationship between changes in unsaturation and fatty acid composition in the regenerating liver were also investigated in this study. Proton nuclear magnetic resonance spectroscopy revealed significantly elevated levels of unsaturation with a maximum on day 5 after partial hepatectomy, compared with untreated controls (11.72±0.55 vs. 11.05±0.26%, P<0.05). No significant changes in unsaturation were found in day 1 regenerating liver, which is rich in absolute amounts of fatty acids. Based on gas-liquid chromatography, the relative amounts of oleic acid (18∶1n−9) and linoleic acid (LA; 18∶2n−6) were increased, while polyunsaturated fatty acids such as arachidonic acid (20∶4n−6) and docosahexaenoic acid (DHA; 22∶6n−3) were decreased on day 1. On the other hand, on day 5 of regeneration, while most fatty acids were returning to their preoperative control levels, only DHA was higher than the control value (7.69±0.58 vs. 5.57±0.37%, P<0.001). The high levels of unsaturation on day 5 were found to be partly due to the increase in DHA. The findings suggest that some significant signals are transmitted during the regeneration process owing to alterations in the membrane structure by the high levels of fatty acid unsaturation and the increase in DHA levels on day 5 after partial hepatectomy.     

Dose-dependent inhibition of cell proliferation induced by lipid peroxidation products in rat hepatoma cells after enrichment with arachidonic acid

Polyunsaturated fatty acids (PUFA) are important constituents of membrane phospholipids, whose levels are decreased in some tumor cells. This deficiency may cause alterations in signal transduction and an interruption of normal cellular events. The enrichment of tumor cells with PUFA may stimulate or inhibit tumor growth, probably depending on the type of PUFA and the cellular concentration of aldehydes derived from restored lipid peroxidation. We examined the effect of several doses of prooxidant on the growth of hepatoma cells with different aldehyde dehydrogenase activities, enriched with arachidonic acid. Two doses of prooxidant were sufficient to reduce growth of hepatoma cells with low aldehyde dehydrogenase activity, whereas three doses were necessary for those with high enzyme activity. In both cases, lipid peroxidation products blocked the cells in the S phase.     

Effect of maternal dietary arachidonic or linoleic acid on rat pup fatty acid profiles

Rapidly growing neonatal mammals accrete relatively large quantities of long chain (≥C₂₀) polyunsaturated fatty acids (LCP) in membrane phospholipids. We have examined accumulation of ω6 LCP in suckling neonatal rat pups during the first 14 d of life when their dams received essential fatty acids in the form of triglycerides containing linoleic acid or arachidonic acid. Dietary levels of these fatty acids were either 1 or 5% of total dietary fatty acids. The fatty acid profile of pup stomach contents (composed solely of the dams' milk) and plasma lipids, as well as liver and brain phospholipids, were determined. Stomach linoleic and arachidonic acid levels reflected the diet of the dams. Pup plasma and liver arachidonic acid levels increased progressively from the group receiving 1% linoleic acid to 5% linoleic acid and from 1% arachidonic acid to 5% arachidonic acid. Interestingly, brain phosphatidylethanolamine and phosphatidylcholine arachidonic acid levels were more stable than plasma or liver levels. These results suggest that the brain may be capable of either selective transport of ω6 LCP or chain elongation/desaturation of linoleic acid. These data indicate that care must be exercised when adding LCP to infant formula since widely divergent accretion rates of arachidonic acid may occur in various tissues.     

Relative incorporation of linoleic and arachidonic acid in phospholipids and triglycerides of different rat tissues

Fat-deficient rats were fed different amounts of methyl linoleate for increasing periods of time. The fatty acid composition of triglycerides and phospholipids of epididymal fat pad, epirenal fat depot, intestinal fat depot, liver, and the pool of heart, kidney, lungs and pancreas was determined. The distribution of the total amount of linoleic and arachidonic acid incorporated into phospholipids and triglycerides per rat was calculated. Phospholipids and triglycerides of depot tissues presented different fatty acid compositions. Although the phospholipids of liver and the pool of heart, kidney, lung and pancreas specifically incorporated linoleic acid at the beginning they very rapidly attained a rather steady composition, whereas triglycerides went on incorporating the acid. The amount of linoleic acid incorporated into the phospholipids of depot tissues was rather small. The triglycerides undoubtedly contributed in the highest proportion to the total pool of linoleic acid. However, the highest proportion of arachidonic acid was found in the total pool of phospholipids. The total amount of linoleic acid incorporated into the phospholipids was an approximately lineal function of the amount of phospholipids independent of period of administration and doses of methyl linoleate. Besides presenting two lineal functions of the amount of phospholipids, arachidonic acid showed a vertical increase coincident with a vertical decrease of the amount of eicosa-5,8,11-trienoic acid. At this period no change in the amount of the phospholipid was shown. This phenomenon is explaioned as a possible direct replacement of eicosatrienoic acid by arachidonic acid.     

Dietary arachidonic acid reduces fatty liver,increases diet consumption and weight gain in ethanol-fed rats

We fed young male Sprague-Dawley rats for 4 wk ad libitum liquid diets containing 34% of the calories as ethanol and 35% as fat with (AA+) and without (AA−) arachidonic acid (20∶4). Additional rats in the control groups were fed similar diets made isocaloric with dextrose with (CA+) and without (CA−) 20∶4. The liver triglyceride (TG) content of rats in the AA+ group was reduced ca. 3-fold over that of rats in the AA-group. The diet consumption and body wts of rats in the AA+ group were significantly greater than those of rats fed alcohol without the 20∶4 supplement (AA−). Also livers from rats in the AA+ group were as large as those from rats in control groups (CA+, CA−) and ca. twice as large as those from rats in the AA-group. The fatty acid composition of liver TG in rats fed the alcohol diet was similar to that of dietary fat. Levels of 20∶4 and docosatetraenoic acid (22∶4) in liver TG fatty acids from rats fed diets without arachidonate (AA−, CA−) were low (trace to 1.6%). After ingestion of arachidonic acid, 20∶4 increased to ca. 10% and 22∶4 to ca. 5%. The content of liver phospholipids was higher in livers of rats fed ethanol (AA−) than in those of controls (CA−). Presented at the ISF/AOCS World Congress, April 27-May 1, 1980, New York City.     

Effects of dietary 9-trans, 12-trans linoleate on arachidonic acid metabolism in rat platelets

In order to determine the minimal amount of dietary 9-trans,12-trans-linoleate which can decrease endoperoxide metabolites synthesized and their precursor in rat platelets, graded amounts (0, 0.1, 0.5, 1.0, 2.5%) of thetrans-linoleate were fed to rats with a constant amount of all-cis-linoleate (2.5%) for 12 weeks. Arachidonic acid levels in platelet phospholipids of groups receiving thetrans-linoleate at 2.5 and 1.0% were significantly (p<0.01) lower than that of the control receiving notrans-linoleate. Concentrations of TXB₂ and PGF_2α in sera of the group receiving 2.5%trans-linoleate were significantly (p<0.05) lower than those of the control; however, there was no difference between the group receiving 1.0%trans-linoleate and the control. To determine whether the difference in serum concentrations of endoperoxide metabolites could be manifested if rats were fed for longer period of time, 2 groups of rats were again fed diets containing 0 and 1.0%trans-linoleate, respectively, for 16 weeks. Arachidonic acid in platelet phospholipids of the group receiving thetrans-linoleate was again significantly (p<0.01) lower than that of the control group. Concentrations of TXB₂ and PGF_2α, and 12-hydroxyeicosatetraenoic acid formed in platelets, were smaller in the group receivingtrans-linoleate than the control group; however, the difference was not statistically significant. These results indicated that all-trans-linoleate can reduce arachidonic acid metabolites formed in rat platelets when its dietary level is equal to or exceeds the level of all-cis-linoeate.     

Fish oil prevents change in arachidonic acid and cholesterol content in rat caused by dietary cholesterol

Rats were fed diets high in either saturated fat (beef tallow) or α-linolenic acid (linseed oil) or eicosapentaenoic and docosahexaenoic acids (fish oil) with or without 2% cholesterol supplementation. Consumption of linseed oil and fish oil diets for 28 days lowered arachidonic acid content of plasma, liver and heart phospholipids. Addition of 2% cholesterol to diets containing beef tallow or linseed oil lowered 20∶4ω6 levels but failed to reduce 20∶4ω6 levels when fed in combination with fish oil. Feeding ω3 fatty acids lowered plasma cholesterol levels. Addition of 2% cholesterol to the beef tallow or linseed oil diet increased plasma cholesterol concentrations but not when fish oil was fed. Feeding the fish oil diet reduced the cholesterol content of liver, whereas feeding the linseed oil diet did not. Dietary cholesterol supplementation elevated the cholesterol concentration in liver in the order: linseed oil > beef tallow > fish oil (8.6-, 5.5-, 2.6-fold, respectively). Feeding fish oil and cholesterol apparently reduced 20∶4ω6 levels in plasma and tissue lipids. Fish oil accentuates the 20∶4ω6 lowering effect of dietary cholesterol and appears to prevent accumulation of cholesterol in plasma and tissue lipids under a high dietary load of cholesterol.     

Vaccenic acid metabolism in the liver of rat and bovine

Hepatic metabolism of vaccenic acid (VA), especially its conversion into CLA, was studied in the bovine (ruminant species that synthesizes CLA) and in the rat (model for nonruminant) by using the in vitro technique of liver explants. Liver tissue samples were collected from fed animals (5 male Wistar rats and 5 Charolais steers) and incubated at 37°C for 17 h under an atmosphere of 95% O₂/5% CO₂ in medium supplemented with 0.75 mM of FA mixture and with 55 μM [1-¹⁴C]VA. VA uptake was about sixfold lower in bovine than in rat liver slices (P<0.01). For both species, VA that was oxidized to partial oxidation products represented about 20% of VA incorporated by cells. The chemical structure of VA was not modified in bovine liver cells, whereas in rat liver cells, 3.2% of VA was converted into 16∶0 and only 0.33% into CLA. The extent of esterification of VA was similar for both animal species (70–80% of incorporated VA). Secretion of VA as part of VLDL particles was very low and similar in rat and bovine liver (around 0.07% of incorporated VA). In conclusion, characteristics of the hepatic metabolism of VA were similar for rat and bovine animals, the liver not being involved in tissue VA conversion into CLA in spite of its high capacity for FA desaturation especially in the rat. This indicates that endogenous synthesis of CLA should take place exclusively in peripheral tissues.     

Effects of oleic,arachidonic and 5,8,11,14-nonadecatetraenoic acids on lipid secretion and ketogenesis in perfused rat liver

The effects of perfused oleic (18∶1n−9), arachidonic (20∶4n−6) and 5,8,11,14-nonadecatetraenoic (19∶4n−5) acids on triglyceride and cholesterol secretion and ketone body production were studied in isolated rat liver. As compared to oleic and 19∶4n−5 acids, both ketone body production and triglyceride secretion were significantly lowered when arachidonic acid was perfused. The concentration of triglyceride in the post-perfused liver was lower upon perfusion with arachidonic acid than upon perfusion with oleic acid or 19∶4n−5 acid. Cholesterol secretion in the liver perfused with arachidonic acid or 19∶4n−5 acid was significantly higher than with oleic acid. The concentration of cholesterol in the post-perfused liver was slightly but significantly higher with 19∶4n−5 acid than with the other fatty acids. The results suggest that 19∶4n−5 acid when compared with arachidonic acid affects lipid metabolism in liver differently.     

藻类花生四烯酸的提取工艺

花生四烯酸作为一种重要的保健、营养品,越来越受到人们的重视。从鱼油、真菌丝、微藻中都可以提取花生四烯酸,但这些工艺都有着明显的缺点。设计了一种从藻类提取花生四烯酸的新工艺过程。     

Ratio of in vivo incorporation of3H arachidonic acid and14C linoleic acid into liver lipids from normal and diabetic rats

Normal and streptozotocin diabetic rats were injected via the portal vein with a labeled solution containing³H arachidonic acid and¹⁴C linoleic acid (³H/¹⁴C ratio, 0.5) during a 1 min period. Livers were quickly frozen, pulverized, and the lipids extracted and fractioned by thin layer chromatography. The incorporation of³H and¹⁴C into liver lipids was measured and the percentage distribution of radioactivity into the different lipid fractions was determined. The incorporation of¹⁴C linoleic acid and³H arachidonic acid into liver lipids is apparently reduced in rats with severe diabetes. The higher³H/^14C ratio found in the 1,2 diglycerides from diabetic rats may be explained by the apparently smaller incorporation of¹⁴C linoleic acid or by an isotopic dilution attributable to the great availability of this acid in diabetic rats. On the other hand, the higher³H/¹⁴C ratio observed in triglycerides and phospholipids from diabetic rats, due to a relatively large incorporation of³H arachidonic acid into this fraction, may be explained by the affinity of the enzymes involved in their synthesis for some 1,2-diglyceride units. Insulin was unable to correct the changes observed in the diabetic rats.     

Inhibition of acid esterase in rat liver by 4,4′-diethylaminoethoxyhexestrol

The effect of 4,4′-diethylaminoethoxyhexestrol (DH) on acid esterase in rat liver was studied in vivo and in vitro. The acid esterase activity in the livers of rats treated with 0.125% DH for 1 week was found to decrease more than 60% as compared with that in untreated rats. The addition of DH to the incubation medium caused considerable inhibition of the acid esterase activity in lysosome from untreated rat liver, and the inhibition type appears to be noncompetitive. The acid lipase activity in rat liver lysosome was also inhibited by DH. Some antihistamic agents and chloroquine also inhibited the acid esterase activity in rat liver lysosome.     

Lipid composition and erucic acid in rat liver cells in culture

Erucic acid (Δ¹³-docosenoic acid) was added to fetal calf serum, then fed to rat liver epithelial cells in culture, and uptake measured at intervals over 24 hr. During the first 6 hr. of incubation, uptake of the docosenoic acid was 21 nmoles/hr/mg protein in 7-day cells, and 15 mmoles/hr/mg protein in 14-day cells. Of¹⁴C-labeled erucic acid taken up by the cells in 24 hr, radioactivity measurements showed 60% of the total lipid¹⁴C activity derived from [1-¹⁴C] 22∶1 in neutral lipid (NL) and 40% in phospholipid (PL); whereas 55% of lipid¹⁴C activity was in NL and 45% in PL when the substrate was [14-¹⁴C] 22∶1. Within the NL fraction, 75% of¹⁴C activity derived from [1-¹⁴C] 22∶1 was in triglyceride (TG) and 11% in cholesterol (CHL), while 79% was in TG and 6.5% in CHL when the substrate was [14-¹⁴C] 22∶1. Triglycerides and cholesteryl esters accumulated in the cells during incubation with erucic acid. Among phospholipids separated by thin layer chromatography, 75% of¹⁴C activity was in lecithin (PC), 10% in phosphatidylethanolamine (PE), 5% in sphingomyelin (SPH), and 1% or less in cardiolipin (DPG). The highest specific activity (SA) was in PC, followed by SPH and PE. Incubation with erucic acid altered fatty acid composition of PC, PE, and SPH, although amounts of phospholipids were unaffected. Gas liquid chromatography analyses detected 18% erucic acid in PC, 2% in PE, and 4–5% in SPH.