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1.
Fatty acid carbons are well-resolved in13C nuclear magnetic resonance (NMR) spectra of lipid extracts, but application of this methodology to the metabolism of13C-labelled fatty acids has not yet been reported. In the present study,13C NMR was used to monitor the presence of 98% [U-13C]eicosapentaenoic acid (EPA) in liver and carcass lipids 24 h after it had been injected into the stomach of a rat. Natural abundance13C NMR spectra of liver total fatty acid extracts were obtained from four control rats for comparison. At 24 h post-injection, quantitative high resolution13C NMR showed13C enrichment in liver fatty acid extracts was present mainly at olefinic and at the n−1 to n−4 carbons, but13C signal intensities for C−1 to C−4 of [U-13C]EPA were markedly reduced or absent. Small13C resonances, possibly indicative of some13C incorporation into docosahexaenoic acid and saturated or monounsaturated fatty acids, were present in spectra of liver fatty acids. Liver and carcass fatty acid composition was similar in both the controls and the EPA-injected rat, suggesting little accumulation of the injected [U-13C]EPA after 24 h. We conclude that the carbon-specific data provided by13C NMR of lipid extracts may be useful in monitoring the fate of individual carbons during tracer studies using13C-labelled fatty acids.  相似文献   

2.
S. G. Miguel 《Lipids》1977,12(12):1080-1083
Slices of rat jejunum were incubated with [2-14C]pyruvate, [1-14C]acetate, or [3H]H2O to determine lipogenic activity. Under all conditions studied, pyruvate acted as a better precursor than acetate for fatty acid synthesis but not for the synthesis of sterol. Exogenous glucose significantly (P≤0.05) increased the conversion of both pyruvate and acetate to fatty acids. By contrast, fasting resulted in a decrease (p≤0.05) in lipogenic activity. The highest levels of lipogenesis were observed when [3H]H2O + glucose at a concentration of 20 mM was used. From such experiments, the absolute rate of fatty acid synthesis in the tissue preparation was calculated: 734±54 nmoles acetyl units incorporated into fatty acids/g tissue/hr.  相似文献   

3.
Kumar D. Mukherjee 《Lipids》1986,21(5):347-352
Lipids in developing seeds ofSinapis alba contain appreciable proportions of (n−7)octadecenoic (vaccenic) acid besides its (n−9) isomer (oleic acid), whereas the constituent very long chain (>C18) monounsaturated fatty acids of these lipids are overwhelmingly composed of the (n−9) isomers. Cotyledons of developingSinapis alba seed use [1-14C]acetate, [1-14C]malonate or [1,3-14C]malonyl-CoA for de novo synthesis of palmitic, stearic and oleic acids and for elongation of preformed oleic, vaccenic and stearic acids to their higher (n−9), (n−7) and saturated homologs, respectively. Moreover, elongation of preformed (n−7)palmitoleic acid to vaccenic acid is observed. Stepwise C2-additions to preformed oleoyl-CoA by acetyl-CoA or malonyl-CoA yielding (n−9)icosenoyl-CoA, (n−9)docosenoyl-CoA and (n−9)tetracosenoyl-CoA are by far the most predominant reactions catalyzed by the elongase system, which seems to have a preference for oleoyl-CoA over vaccenoyl-CoA as the primer. The pattern of14C-labeling of the very long chain fatty acids formed from either acetate or malonate shows a close analogy in the mode of elongation of monounsaturated and saturated fatty acids.  相似文献   

4.
Our objective was to develop a suitable probe to study metabolism of polyunsaturated fatty acids by 13C nuclear magnetic resonance (NMR) in the suckling rat pup. [3-13C] γ-Linolenic acid was chemically synthesized, and a 20 mg (Experiment 1) or 5 mg (Experiment 2) dose was injected into the stomachs of 6–10-day-old suckling rat pups that were then killed over a 192 h (8 d) time course. 13C NMR showed that 13C in γ-linolenate peaked in liver total lipids by 12-h post-dosing and that [5-13C]-arachidonic acid peaked in both brain and liver total lipids 48–96 h post-dosing. 13C enrichment in brain γ-linolenic acid was not detected by NMR, but gas chromatography-combustion-isotope ratio mass spectrometry showed that its mass enrichment in brain phospholipids at 48–96 h post-dosing was 1–2% of that in brain arachidonic acid. 13C was present in liver and brain cholesterol and in perchloric acid-extractable water-soluble metabolites in the brain, liver and carcass. We conclude that low but measurable amounts of exogenous γ-linolenic acid do access the suckling rat brain in vivo. The slow time course of [5-13C] arachidonic acid appearance in the brain suggests most of it was probably transported there after synthesis elsewhere, probably in the liver. Some carbon from γ-linolenic acid is also incorporated into lipid products other than n−6 long-chain polyunsaturated fatty acids.  相似文献   

5.
Hepatocytes isolated from female rats meal-fed a high-glucose diet were incubated in Krebs-Henseleit bicarbonate medium containing 16.5 mM glucose,3H2O, and14C-labeled amino acids (−)-Hydroxycitrate depressed the incorporation of3H2O and [14C] alanine into fatty acids and cholesterol. Incorporation of [U-14C] leucine into lipids was not affected but incorporation of3H2O into lipids was decreased significantly by (−)-hydroxycitrate. (−)-Hydroxycitrate depressed the incorporation of radioactivity from [2-14C]leucine into fatty acids and cholesterol by 61 and 38%, respectively, and stimulated the incorporation of radioactivity from [4,5-3H]leucine 35 and 28%. As [2-14C]leucine labels the acetyl-CoA pool and [4,5-3H]leucine labels the acetoacetate pool, it was concluded that mitochondrial 3-hydroxy-3-methylglutaryl-CoA is not incorporated intact into cholesterol, and that acetoacetate can be activated effectively in the liver cytosol for support of cholesterol and fatty acid synthesis.  相似文献   

6.
The biosynthesis of (Z,Z)-6,9-heptacosadiene, the major cuticular hydrocarbon component of the American cockroach, was examined by radiotracer and13C-nuclear magnetic resonance (NMR) techniques. Sodium [1-14C] acetate was incorporated about equally into the saturated and diunsaturated hydrocarbons, whereas [1-14C] linoleate preferentially labeled the C27 alkadiene and [9,10-3H] oleate labeled the C27 alkadiene almost exclusively.13C-NMR demonstrated that [2-13C] acetate labeled carbons 25 and 27 but not carbon 3 of the C27 alkadiene. In addition, ozonolysis of the diene labeled from [1-14C] acetate followed by radio-gas liquid chromatography showed that carbons 1–6 were not labeled, whereas the fragment containing carbons 10–27 was labeled. The data presented in this paper indicate that linoleate from the diet or synthesized de novo is elongated by the addition of acetate units and is then decarboxylated.  相似文献   

7.
The gastrointestinal handling and metabolism of [1-13C]palmitic acid given as the free fatty acid was examined in six healthy women by measuring the excretion of13C-label in stool and in breath as13CO2. The gastrointestinal handling of [1-13C]palmitic acid was compared with the apparent absorption of dietary lipid by measuring lipid losses in stool. The variation both within and between subjects was determined by repeating the study in the same individuals on separate occasions. The time course for excretion of label in stool over the five-day study period followed a common pattern, with most of the label excreted over the first two days of the stool collection.13C-Label excreted in stool over the five-day study period was 14.3±9.8% of that administered and on repeating the trial was 31.6±24.7% (not significantly different due to variability); there was poor agreement within subjects. Lipid excreted in stool expressed as a percentage of ingested lipid was 5.2±4.4% in Trial 1 and 5.9±4.0% in Trial 2, and was the same in each individual on repeating the trial. There was no clear relationship between the excretion of13C-label and lipid in stool (Trial 1:R=−0.43,P>0.40; Trial 2:R=−0.02,P>0.97). On the first occasion, 22.0±4.5% of the administered label was excreted on breath over the 15-h study period and on repeating the trial was 15.8±9.5% (not significantly different) with poor repeatability in a given individual. There was an inverse relationship between the proportion of13C-label excreted in stool and that excreted on breath in Trial 1 (R=−0.80,P>0.06) with a weaker association observed in Trial 2 (R=−0.49,P>0.32). Correcting for differences in the apparent absorption of label reduced the variability in its excretion in breath observed between subjects, particularly in Trial 2. It is concluded that although there are differences in the gastrointestinal handling of [1-13C]palmitic acid both within and between healthy adults, the postprandial oxidation of absorbed substrate was similar. The assumptions underlying these observations need to be examined by characterizing the nature of13C-label in stool.  相似文献   

8.
The additional mass associated with 13C in metabolic tracers may interfere with their metabolism. The comparative isomerization and biohydrogenation of oleic, [1-13C]oleic, and [U-13C]oleic acids by mixed ruminal microbes was used to evaluate this effect. The percent of stearic, cis-14 and- 15, and trans-9 to-16 18∶1 originating from oleic acid was decreased for [U-13C]oleic acid compared with [1-13C]oleic acid. Conversely, microbial utilization of [U-13C]oleic acid resulted in more of the 13C label in cis-9 18∶1 compared with [1-13C]oleic acid (53.7 vs. 40.1%). The isomerization and biohydrogenation of oleic acid by ruminal microbes is affected by the mass of the labeled tracer.  相似文献   

9.
R. P. Bird  H. H. Draper 《Lipids》1982,17(8):519-523
Primary cultures of rat skin fibroblasts were used as a model system to investigate the cellular uptake and oxidation of malonaldehyde (MA). The cells were grown in a medium containing 10−5 M, 10−4 M or 10−3 M concentrations of [1,3-14C]MA. There was a limited, concentration-dependent uptake of MA by 24 hr (∼4% at all concentrations). The uptake of [1,2-14C]acetate by 24 hr was ∼24%; 83–89% of the14C in the MA taken up was oxidized to14CO2 by 24 hr and ∼5% was recovered in the major lipids. Despite its low uptake and rapid oxidation to CO2, pretreatment of the cells with 10−3 M MA for 24 hr produced a latent inhibition of [14C]glucose oxidation. Limited cellular uptake of MA may explain the tolerance of cells grown in culture to relatively high MA concentrations.  相似文献   

10.
Oleate is one of the most abundant dietary fatty acids, and much remains to be learned about its metabolism in fat cells. We studied the incorporation of exogenous [1-13C]-oleate into triglycerides (TG) in differentiating 3T3L1 preadipocytes using 13C NMR spectroscopy. The quantity of oleate incorporated into TG was found to increase as preadipocytes differentiated into fat cells. The ratio of unesterified [1-13C]oleate to total stored fatty acids was higher in less differentiated cells, and declined at later stages of differentiation as cells accumulated fatty acids through de novo synthesis. When added as the only exogenous fatty acid, oleate was largely esterified at the sn-2 position. When equimolar unlabeled linoleate was co-provided at the same time, the ratio of [1-13C]oleate esterified at the sn-1,3 position increased, implying competition between linoleate and oleate for esterification, especially at the sn-2 position. When cells pre-enriched with [1-13C]oleate (esterified to TG) were treated with isoproterenol, a lipolytic agent, most of the [1-13C]oleate was still found in TG, despite a high rate of lipolysis determined by measuring glycerol release. This implies extensive re-esterification of the oleate released by lipolysis.  相似文献   

11.
Marine fish have an absolute dietary requirement for C20 and C22 highly unsaturated fatty acids. Previous studies using cultured cell lines indicated that underlying this requirement in marine fish was either a deficiency in fatty acyl Δ5 desaturase or C18–20 elongase activity. Recent research in turbot cells found low C18–20 elongase but high Δ5 desaturase activity. In the present study, the fatty acid desaturase/elongase pathway was investigated in a cell line (SAF-1) from another carnivorous marine fish, sea bream. The metabolic conversions of a range of radiolabeled polyunsaturated fatty acids that comprised the direct substrates for Δ6 desaturase ([1-14C]18∶2n−6 and [1-14C]18∶3n−3), C18–20 elongase ([U-14C]18∶4n−3), Δ5 desaturase ([1-14C]20∶3n−6 and [1-14C]20∶5n−3), and C20–22 elongase ([1-14C]20∶4n−6 and [1-14C]20∶5n−3) were utilized. The results showed that fatty acyl Δ6 desaturase in SAF-1 cells was highly active and that C18–20 elongase and C20–22 elongase activities were substantial. A deficiency in the desaturation/elongation pathway was clearly identified at the level of the fatty acyl Δ5 desaturase, which was very low, particularly with 20∶4n−3 as substrate. In comparison, the apparent activities of Δ6 desaturase, C18–20 elongase, and C20–22 elongase were approximately 94-, 27-, and 16-fold greater than that for Δ5 desaturase toward their respective n−3 polyunsaturated fatty acid substrates. The evidence obtained in the SAF-1 cell line is consistent with the dietary requirement for C20 and C22 highly unsaturated fatty acids in the marine fish the sea bream, being primarily due to a deficiency in fatty acid Δ5 desaturase activity.  相似文献   

12.
Treatment of 1-β-(4′-acetylphenyl)vinyl-3-vinyl-1,1,3,3-tetramethyldisiloxane (I) (an AB2 monomer) with dihydridocarbonyltris(triphenylphosphine)ruthenium (Ru) leads to a hyperbranched material, poly[1-β-(4′-acetylphenyl)vinyl-3-vinyl-1,1,3,3-tetramethyldisiloxane] (II). I has been prepared by a Pd catalyzed Heck reaction between 4-bromo-acetophenone and 1,3-divinyl-1,1,3,3-tetramethyldisiloxane. The structure of the soluble hyperbranched material (II) has been determined by 1H, 13C and 29Si NMR, as well as by IR and UV spectroscopy. It has also been characterized by GPC, TGA, DSC and elemental analysis. Polymerization occurs by Ru catalyzed addition of the aromatic C−H bonds which are ortho to the activating acetyl group of I across the C−C double bond of the terminal Si-vinyl group in an anti-Markovnikov manner. Received: 8 September 1997/Revised version: 19 October 1997/Accepted: 20 November 1997  相似文献   

13.
(−)-Hydroxycitrate and (+)-allo-hydroxycitrate were investigated for their effects on lipid synthesis in vivo under conditions of either high carbohydrate feeding or 24 hr fasting. Changes in rates of lipid synthesis resulting from the oral administration of these compounds were monitored with the use of radiolabeled H2O, alanine, and acetate. In the fed rat, (−)-hydroxycitrate significantly reduced the incorporation of H2O and alanine into fatty acids and cholesterol. An increased incorporation of labeled H2O into fatty acids but no change in cholesterol synthesis in the fasted rat suggested that (−)-hydroxycitrate may be an activator of acetyl CoA carboxylase. With (−)-hydroxycitrate administration, acetate incorporation into fatty acids and cholesterol was subject to pool dilution effects under fed or fasted states. (+)-allo-Hydroxycitrate was ineffective in modulating the rates of fatty acid synthesis under either nutritional condition. Both (−)-hydroxycitrate and (+)-allo-hydroxycitrate were shown to be in vitro activators of acetyl CoA carboxylase, the former being a much stronger activator than the latter. Thus, stereospecificity of the hydroxycitrate isomers was demonstrated in both the inhibition of lipid synthesis (previously shown to occur at adenosine triphosphate citrate lyase) and the stimulation of fatty acid synthesis (possibly occurring at acetyl CoA carboxylase).  相似文献   

14.
Yu-Yan Yeh  Shaw-Mei Yeh 《Lipids》1994,29(3):189-193
Prompted by the reported hypolipidemic activity of garlic, the present study was undertaken to elucidate the mechanism(s) underlying the cholesterol-lowering effects of garlic. Rat hepatocytes in primary culture were used to determine the short-term effects of garlic preparations on [1-14C]acetate and [2-3H]glycerol incorporation into cholesterol, fatty acids and glycerol lipids. When compared with the control group, cells treated with a high concentration of garlic extracts [i.e., petroleum ether- (PEF), methanol- (MEF) and water-extractable (WEF) fractions from fresh garlic] showed decreased rates of [1-14C]acetate incorporation into cholesterol (by 37–64%) and into fatty acids (by 28–64%). Kyolic containingS-allyl cysteine and organosulfur compounds inhibited cholesterogenesis in a concentration dependent manner with a maximum inhibition of 87% at 0.4 mM. At this concentration, Kyolic decreased [1-14C]acetate incorporation into fatty acids by 67%.S-allyl cysteine at 2.0 and 4.0 mM inhibited cholesterogenesis by 20–25%. PEF, MEF and WEF depressed the rates of [2-3H]glycerol incorporation into triacylglycerol, diacylglycerol and phospholipids in the presence of acetate, but not in the presence of oleate. The results suggest that the hypocholesterolemic effect of garlic stems, in part, from decreased hepatic cholesterogenesis, whereas the triacylglycerol-lowering effect appears to be due to inhibition of fatty acid synthesis. Primary hepatocyte cultures as used in the present study have been proven useful as tools for screening the anticholesterogenic properties of garlic principles.  相似文献   

15.
Liver slices, from mice fasted for one day and then refed for three days either a 15% corn oil diet or a 15% corn oil diet containing eicosa-5,8,11,14-tetraynoic acid (TYA), were incubated with [1-14C] acetate or [3H]H2O to determine lipogenic capacity. Dietary TYA produced a twofold stimulation in fatty acid and cholesterol synthesis. TYA also caused an increase in the relative proportion of linoleate (C18∶2) and a decrease in that of arachidonate (C20∶4) in liver. Thus, (a) despite high levels of C18∶2, hepatic lipogenesis can be increased, and (b) even short term feeding of TYA can alter the hepatic fatty acid composition presumably by inhibition of arachidonate synthesis from linoleate.  相似文献   

16.
Oil palm (E. guineensis) fruits at three stages of development were studied. At week 12–13 after anthesis, the endosperm had started accumulating oil and tissue slices incorporated [1-14C] acetate into fatty acids which resembled those found in the mature endosperm. The mesocarp contained very little oil and incorporated acetate into polar lipids. At week 16–17, the mesocarp started to accumulate oil; this was reflected in the [14C] lipid products from acetate incubation. At or just prior to this stage, an increase in the endogenous linoleic and linolenic acid content and the increase in fruit size indicated cellular growth in the mesocarp tissue. At week 20–21 the fruit was ripe, and both endosperm and mesocarp tissues were filled with storage oil. [14C] Fatty acids synthesized from acetate by mesocarp slices at this stage were the same as the endogenous storage fatty acids in bothE. guineensis andE. oleifera. A very weak fatty acid synthesizing activity was seen in the mature endosperm, but the products had no relationship to the storage lipid.  相似文献   

17.
Jen-sie Tou 《Lipids》1986,21(5):324-327
Docosahexaenoic acid was not only acylated into phospholipids but also into triacylglycerols by intact human neutrophils. The distribution of radiolabeled docosahexaenoic acid among individual phospholipids was dependent on the incubation time. [1-14C]Docosahexaenoic acid at all concentrations (1 to 8 μM) was acylated mainly into phosphatidic acid after 1–2 min incubation, and the radioactivity of phosphatidic acid started to decline after a longer period of incubation, suggesting the participation of docosahexaenoyl-phosphatidic acid in the synthesis of other glycerolipids. It was acylated primarily into phosphatidylcholine (PC) and phosphatidylethanolamine (PE) after a 2-hr incubation. The labeled phosphatidic acid may be rapidly deacylated and the 22∶6(n−3) moiety is then reacylated into other lysophospholipids. The low levels of [14C]22∶6(n−3) in 1,2-diacylglycerol suggest that the deacylation-reacylation cycle may be a major pathway in the formation of [14C]22∶6(n−3)-PC and-PE in intact neutrophils. This n−3 fatty acid was a relatively poor substrate for acylation into phosphatidyl-inositol as compared to arachidonic acid and eicosapentaenoic acid. However, the patterns of distribution of all three polyunsaturated fatty acids among the diacyl-and ether-linked class compositions of PC and PE were similar. These data suggest the potential of increasing the content of docosahexaenoic acid of membrane lipids in neutrophils by dietary supplement of this fatty acid.  相似文献   

18.
The biosynthesis of fatty acids in the diatomPhaeodactylum tricornutum was studied. The diatom was incubated with sodium [114C] acetate and the acids [1-14C] palmitic, [1-14C] stearic, [1-14C] linoleic and [1-14C] α-linolenic. The distribution of radioactivity in the products was determined by gas liquid radiochromatography. The diatom synthesized “de novo” not only saturated and monounsaturated fatty acids, but also linoleic, α-linolenic and other fatty acids including the highly polyunsaturated 20∶5ω3 and 22∶6ω3. When labeled acetate, stearic, α-linolenic or even linoleic acid were incubated with the diatom, the polyunsaturated C20 fatty acids synthesized belonged predominantly to the ω 3 family. The existence of Δ9, Δ6, Δ5, Δ4, ω6 and possibly ω3 desaturases inP. tricornutum is suggested. Member of the Carrera del Investigador Científico of the Comisión de Investigaciones Científicas de la Provincia de Buenos Aires. Member of the Carrera del Investigador Cientifico of the Consejo Nacional de Investigaciones Cientificas y Técnicas.  相似文献   

19.
Ves-Losada A  Maté SM  Brenner RR 《Lipids》2001,36(3):273-282
Liver nuclear incorporation of stearic (18∶0), linoleic (18∶2n−6), and arachidonic (20∶4n−6) acids was studied by incubation in vitro of the [1-14C] fatty acids with nuclei, with or without the cytosol fraction at different times. The [1-14C] fatty acids were incorporated into the nuclei as free fatty acids in the following order: 18∶0>20∶4n−6≫18∶2n−6, and esterified into nuclear lipids by an acyl-CoA pathway. All [1-14C] fatty acids were esterified mainly to phospholipids and triacylglycerols and in a minor proportion to diacylglycerols. Only [1-14C] 18∶2n−6-CoA was incorporated into cholesterol esters. The incorporation was not modified by cytosol addition. The incorporation of 20∶4n−6 into nuclear phosphatidylcholine (PC) pools was also studied by incubation of liver nuclei in vitro with [1-14C]20∶4n−6-CoA, and nuclear labeled PC molecular species were determined. From the 15 PC nuclear molecular species determined, five were labeled with [1-14C]20∶4n−6-CoA: 18∶0–20∶4, 16∶0–20∶4, 18∶1–20∶4, 18∶2–20∶4, and 20∶4–20∶4. The highest specific radioactivity was found in 20∶4–20∶4 PC, which is a minor species. In conclusion, liver cell nuclei possess the necessary enzymes to incorporate exogenous saturated and unsaturated fatty acids into lipids by an acyl-CoA pathway, showing specificity for each fatty acid. Liver cell nuclei also utilize exogenous 20∶4n−6-CoA to synthesize the major molecular species of PC with 20∶4n−6 at the sn-2 position. However, the most actively synthesized is 20∶4–20∶4 PC, which is a quantitatively minor component. The labeling pattern of 20∶4–20∶4 PC would indicate that this molecular species is synthesized mainly by the de novo pathway.  相似文献   

20.
The de novo biosynthesis of fatty acids in the wild, calanoid copepodParacalanus parvus was studied. The incubation of labeled acetate proved the de novo biosynthesis of saturated and monounsaturated even fatty acids from 14 to 20 carbons and the 22∶1 acid. Saturated and monounsaturated uneven fatty acids from 15 to 21 carbons were also synthesized. The copepod could not synthesize linoleic and α-linolenic acids. By administration of [1-14C]palmitate, [1-14C] stearate and [1-14C]oleate, it was possible to elucidate the general pattern of the de novo biosynthesis of fatty acids in the wildP. parvus.  相似文献   

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