0000000000008090
AUTHOR
Jean Bézard
Influence of cyclopropene fatty acids (Baobab seed oil) feeding on the in vitro Δ9 desaturation of stearic acid in rat liver microsomes
Abstract Δ9 desaturation of [1−14C] stearic acid to oleic acid was studied in liver microsomes of rats fed either fresh Baobab seed oil, containing cyclopropene fatty acids (malvalic acid and sterculic acids), or heated Baobab oil, containing only minor amounts of these acids. This was compared with rats fed a control oil mixture or a fat-free diet. Up to 50 nmol of substrate, Δ9 desaturation rate was the same in rats fed the three fat diets. From 50 to 150 nmol, the desaturation rates were practically constant in rats fed fresh Baobab seed oil. It increased in heated oil-fed rats at the same rate as in the control group, but more slowly than in the fat-free diet group. Fatty acid compositi…
Distribution of Δ5-olefinic acids in the triacylglycerols fromPinus koraiensisandPinus pinasterseed oils
Purified triacylglycerols (TAG) fromPinus koraiensis andP. pinaster seed oils, which are interesting and commercially available sources of Δ5-olefinic acids (i.e.,cis-5,cis-9,cis-12 18:3 andcis-5,cis-11,cis-14 20:3 acids) were fractionated by reversed-phase high-performance liquid chromatography, and each fraction was examined by capillary gas-liquid chromatography for its fatty acid composition. A structure could be assigned to more than 92% of TAG from both oils. In both instances, ca. 48% of the TAG were shown to contain at least one δ5-olefinic acid. In the great majority of TAG, our data showed that there is only one molecule of δ5-olefinic acid per molecule of TAG. This is compatible …
Age-related changes in delta 6 and delta 5 desaturase activities in rat liver microsomes.
Age-related changes in Δ6 desaturation of [1-14C]α-linolenic acid and [1-14C]linoleic acid and in Δ5 desaturation of [2-14C]dihomo-γ-linolenic acid were studied in liver microsomes from Wistar male rats at various ages ranging from 1.5 to 24 mon. Desaturase activities were expressed both as specific activity of liver microsomes and as the capacity of whole liver to desaturate by taking into account the total amount of liver microsomal protein. Δ6 Desaturation of α-linolenic acid increased from 1.5 to 3 mon and then decreased linearly up to 24 mon to reach the same desaturation capacity of liver measured at 1.5 mon. The capacity of liver to desaturate linoleic acid increased up to 6 mon and …
Pathway of alpha-linolenic acid through the mitochondrial outer membrane in the rat liver and influence on the rate of oxidation. Comparison with linoleic and oleic acids.
The movement of alpha-linolenic acid (C18:3, n-3) through the mitochondrial outer membrane to oxidation sites was studied in rat liver and compared with the movement of linoleic acid (C18:2, n-6) and oleic acid (C18:1, n-9). All differ in the degree of unsaturation, but have the same chain length and the same position of the first double bond when counted from the carboxyl end. The following results were obtained. (1) The overall beta-oxidation in total mitochondria was in the order C18:3, n-3 greater than C18:2, n-6 greater than C18:1, n-9, independent of the amount of albumin in the medium. (2) The rate of formation of acylcarnitine from acyl-CoA was higher with oleoyl-CoA than with linol…
Effect of low levels of dietary fish oil on fatty acid desaturation and tissue fatty acids in obese and lean rats
The effect of very low levels of dietary long-chain n-3 fatty acids on delta 6 desaturation of linoleic acid (18:2n-6) and alpha-linolenic acid (18:3n-3), and on delta 5 desaturation of dihomo-gamma-linolenic acid (20:3n-6), in liver microsomes and its influence on tissue fatty acids were examined in obese and lean Zucker rats and in Wistar rats. Animals fed for 12 wk a balanced diet containing ca. 200 mg of long-chain polyunsaturated n-3 fatty acids per 100 g of diet were compared to those fed the same amount of alpha-linolenic acid. Low amounts of long-chain n-3 fatty acids greatly inhibited delta 6 desaturation of 18:2n-6 and delta 5 desaturation of 20:3n-6, while delta 6 desaturation of…
Effect of simvastatin on desaturase activities in liver from lean and obese zucker rats
The effect of simvastatin, a hypocholesterolemic drug, on the biosynthesis of arachidonic acid was studied in obese and lean Zucker rats. After administration of 2 mg/kg body weight/d for 13 d, delta 6 and delta 5 desaturase activities were measured in liver microsomes at two substrate concentrations. In untreated rats, the delta 6 desaturation rate was similar in the obese and lean rats when measured at saturating substrate levels, whereas delta 5 desaturation was lower in the obese animals. Treatment with simvastatin did not change delta 6 desaturation in either phenotype but increased delta 5 desaturation in obese rats to reach the unchanged rate observed in lean animals. The changes wer…
Short term treatment by fenofibrate enhances oxidative activities towards longchain fatty acids in the liver of lean zucker rats
Lean Zucker rats were dosed orally for 1 week with fenofibrate (100 mg/kg/day). Liver weights of treated rats, expressed as per cent of body weight, were increased, while protein, DNA and triacylglycerol contents were not changed to any great extent per gram of liver, but increased when expressed per whole liver. Compared with the control animals, activities of fatty acid oxidase, of the peroxisomal fatty acid-oxidizing system and of catalase were markedly enhanced by fenofibrate, both per gram of liver and per total liver, while urate oxidase activity was slightly depressed when expressed per gram of liver. The activity of cytochrome c oxidase used as a mitochondrial marker was only higher…
EFFECTS OF CYCLOPROPENOID FATTY ACIDS (BAOBAB SEED OIL) ON THE FATTY ACID PROFILE OF LIPIDS FROM DIFFERENT TISSUES IN THE RAT
Rats were fed for 8 weeks a diet containing either Baobab seed oil (FBO) rich in cyclopropenoid fatty acids (CPFA) or heated oil (HBO) devoid of CPFA. In FBO rats, compared to controls (CO), the monounsaturated fatty acid content of adipose tissue lipids, liver triacylglycerols, liver and adrenal cholesteryl esters and liver, kidney, adrenal phospholipids were highly decreased to the benefit of saturated fatty acid content. In cholesteryl esters and phospholipids, the arachidonic acid (20:4 n-6) content was markedly decreased, especially in adrenals. In adrenal cholesteryl esters, adrenic acid (22:4 n-6) content was decreased more than its precursor. The effects of HBO on the tissue lipid f…
Analysis and fractionation of natural source diacylglycerols as urethane derivatives by reversed-phase high-performance liquid chromatography
Abstract Reversed-phase high-performance liquid chromatography on a thermostatted octadecylsilyl column was used to separate and fractionate mixtures of diacylglycerols after derivatization with 3,5-dinitrophenyl isocyanate (urethane derivatives). In addition to the separation of commercial diacylglycerol species, the separation of diacylglycerols obtained from peanut oil and cottonseed oil triacylglycerols by chemical hydrolysis is reported. Acetonitrile-acetone mixtures were used for elution of the diacylglycerol urethane derivatives. Unsaturated and saturated derivatives were detected by their refractive indices. They were then collected and their fatty acids analysed as methyl esters by…
Method for evaluating the bioconversion of radioactive polyunsaturated fatty acids by use of reversed-phase liquid chromatography
Abstract Reversed-phase high-performance liquid chromatography on a thermostatted octadecylsilyl column was used to separate mixtures of labelled polyunsaturated fatty acids (as their methyl esters) formed by successive desaturations and elongations of labelled linoleic (18:2 n - 6) or linolenic (18:3 n -3) acid by rat liver microsomes. Acetonitrile-water mixtures were used for elution of the esters. Unsaturated and saturated esters were detected by their refractive indices. The order of elution of fatty acid methyl esters in complex mixtures varies as a function of the chain length and unsaturation, analysis temperature, water concentration and solvent flow-rate. The peak areas vary as a f…
Delta 5-desaturation of dihomogammalinolenic acid (20:3(n-6)) into arachidonic acid (20:4(n-6)) by rat liver microsomes and incorporation of fatty acids in microsome phospholipids.
Abstract Liver microsomes of rats fed an essential fatty acid (EFA)-deficient diet or a commercial balanced diet were used to study the effect of incubation time on the Δ 5 -desaturation of [ 14 C] dihomogammalinolenic acid (20:3( n −6)) into arachidonic acid (20:4( n −6)) and incorporation of the two acids into microsomal phospholipids. The EFA-deficient diet highly increased the desaturation rate of 20:3( n −6). Incorporation of the formed 20:4( n −6) into microsomal phospholipids was also increased but at saturating concentration of substrate only. At early times of incubation, the precursor 20:3( n −6) was rapidly incorporated into phospholipids. Formation and incorporation of 20:4( n −…
Effect of dietary n−3 and n−6 polyunsaturated fatty acids on lipid-metabolizing enzymes in obese rat liver
This study was designed to examine whether n-3 and n-6 polyunsaturated fatty acids at a very low dietary level (about 0.2%) would alter liver activities in respect to fatty acid oxidation. Obese Zucker rats were used because of their low level of fatty acid oxidation, which would make increases easier to detect. Zucker rats were fed diets containing different oil mixtures (5%, w/w) with the same ratio of n-6/n-3 fatty acids supplied either as fish oil or arachidonic acid concentrate. Decreased hepatic triacylglycerol levels were observed only with the diet containing fish oil. In mitochondrial outer membranes, which support carnitine palmitoyltransferase I activity, cholesterol content was …
Enantiomer separation by chiral-phase liquid chromatography of urethane derivatives of natural diacylglycerols previously fractionated by reversed-phase liquid chromatography
Abstract Enantiomers of diacylglycerols such as 3,5-dinitrophenyl isocyanate (urethane) derivatives previously fractionated by reversed-phase high-performance liquid chromatography (HPLC) were separated by HPLC on a chiral phase column [N-( R )-1-(α-naphthyl)ethylamino-carbonyl-( S )-valine chemically bound to γ-aminopropylsilanized silica]. In addition to the separation of commercial monoacid-diacylglycerol isomers, separation of diacid-diacylglycerol isomers obtained from peanut oil and cottonseed oil triacylglycerols by chemical hydrolysis is reported. Hexane-ethylene dichloride-ethanol mixtures were used for elution of the diacylglycerol derivative isomers, which were detected by their …
Dietary arachidonic acid and hepatic desaturation of fatty acids in obese zucker rats
The effect of low levels of dietary arachidonic acid (20:4n-6) on Δ6 desaturation of linoleic acid (18:2n-6) and α-linolenic acid (18:3n-3), and on Δ5 desaturation of dihomo-γ-linolenic acid (20:3n-6) were studied in liver microsomes of obese Zucker rats, in comparison with their lean littermates. Fatty acid composition of serum total lipids and of phospholipids from liver microsomes and from total heart and kidney was determined to see whether modifications of desaturation rate, if any, were reflected in the tissue fatty acid profiles. Animals fed for 12 wk on a balanced diet, containing 20:4n-6 and 18:2n-6, were compared to those fed 18:2n-6 only. The low amount of dietary 20:4n-6 greatly…
Inhibition of fatty acid delta 6- and delta 5-desaturation by cyclopropene fatty acids in rat liver microsomes.
delta 6-Desaturation of linoleic acid and delta 5-desaturation of dihomo-gamma-linolenic acid were measured in liver microsomes from rats fed fresh Baobab seed oil containing cyclopropene fatty acids (malvalic acid and sterculic acid) or heated Baobab seed oil practically devoid of these fatty acids or control oil. The presence of cyclopropene fatty acids in the fresh Baobab oil diet highly depressed both desaturations, but delta 6- more than delta 5-desaturation. The decreased capacity of microsomes to desaturate was reflected in the lower arachidonic acid content in microsomal phospholipids from rats fed this oil. However it was also lower in rats fed heated oil although in vitro delta 6-…
Involvement of microsomal vesicles in part of the sensitivity of carnitine palmitoyltransferase I to malonyl-CoA inhibition in mitochondrial fractions of rat liver.
Liver mitochondrial fractions as normally isolated contain only 10-20% of total mitochondria and may not be representative of the whole mitochondrial population. This study was designed to evaluate the dependence of the sensitivity of carnitine palmitoyl-transferase I (CPT I) to malonyl-CoA inhibition in mitochondrial fractions that are not normally studied. Four fractions prepared from rat liver were found to be contaminated to different extents by microsome vesicles, on the basis of marker-enzyme activities and micrographic data. Purification of mitochondrial fractions on a Percoll gradient decreased to some extent the microsomal contamination, which was due in part to the existence of cl…
Enhancement of activities relative to fatty acid oxidation in the liver of rats depleted of l-carnitine by d-carnitine and a γ-butyrobetaine hydroxylase inhibitor
Abstract This study was designed to examine whether the depletion of l -carnitine may induce compensatory mechanisms allowing higher fatty acid oxidative activities in liver, particularly with regard to mitochondrial carnitine palmitoyltransferase I activity and peroxisomal fatty acid oxidation. Wistar rats received d -carnitine for 2 days and 3-(2,2,2-trimethylhydrazinium)propionate (mildronate), a non-competitive inhibitor of γ-butyrobetaine hydroxylase, for 10 days. They were starved for 20 hr before being sacrificed. A dramatic reduction in carnitine concentration was observed in heart, skeletal muscles and kidneys, and to a lesser extent, in liver. Triacylglycerol content was found to …
The metabolism and availability of essential fatty acids in animal and human tissues.
Essential fatty acids (EFA), which are not synthesized in animal and human tissues, belong to the n-6 and n-3 families of polyunsaturated fatty acids (PUFA), derived from linoleic acid (LA, 18:2n-6) and alpha-linolenic acid (LNA, 18:3n-3). Optimal requirements are 3-6% of ingested energy for LA and 0.5-1% for LNA in adults. Requirements in LNA are higher in development. Dietary sources of LA and LNA are principally plants, while arachidonic acid (AA, 20:4n-6) is found in products from terrestrian animals, and eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) are found in products from marine animals. EFA are principally present in dietary triacylglycerols, which should be hydrolyze…
Elongation and desaturation of arachidonic and eicosapentaenoic acids in rat liver. Effect of clofibrate feeding
The fatty acid elongation-desaturation ability of 5,8,11,14-eicosatetraenoic (20:4(n-6)) and 5,8,11,14,17-eicosapentaenoic (20:5(n-3)) acids was determined in both liver microsomal and light mitochondrial (rich in peroxisomes) fractions of untreated and clofibrate treated rats. The elongation and the subsequent desaturation steps were performed in the corresponding favorable media. 20:5(n-3) elongation was about 2-times more extensive than that of 20:4(n-6). Clofibrate feeding for 10 days resulted in a marked decrease in the elongation rate with the two substrates, while the delta 4 desaturation rate was increased. There were small differences in the elongation rate between the microsomal a…
Determination of molecular species of oil triacylglycerols by reversed-phase and chiral-phase high-performance liquid chromatography
A method is described for the determination of molecular species of oil triacylglycerols. The method is based on the analytical separation of the enantiomericsn-1,2-andsn-2,3-diacylglycerols, derived from triacylglycerols, by high-performance liquid-chromatography (HPLC) on a chiral column containing N-(R)-1-(α-naphthyl)ethylaminocarbonyl-(S)-valinecarbonyl-(S)-valine as stationary phase. Model triacyl-glycerol molecules comprising three known fatty acids were isolated from peanut oil and cottonseed oil by a combination of argentation-TLC and reversed-phase HPLC and submitted to partial chemical deacylation. The derivedsn-1,2(2,3)-diacylglycerols were analyzed and fractionated as 3,5-dinitr…
Separation of monoacylglycerols by reversed-phase high-performance liquid chromatography
Abstract Reversed-phase high-performance liquid chromatography on a thermostated octadecylsilyl column was used to separate complex mixtures of monoacyglycerols formed by chemical deacylation of natural triacyclycerols. Acetonitrile-water mixtures were used for elution of the underivatized monoacylglycerols and a different refractometer was used for their detection. The order of elution of monoacylglycerols in complex mixtures varies as a function of the chain length, unsaturation, positional isomerism, analysis temperature and eluent water concentration. The peak areas were representative of the amounts of monoacylglycerols detected. The method can be applied to compositional analysis and …
Polyunsaturated n-3 and n-6 fatty acids at a low level in the diet alter mitochondrial outer membrane parameters in Wistar rat liver
Abstract This study was designed to examine whether n-3 and n-6 polyunsaturated fatty acids (PUFA) at a very low level in the diet (about 0.2%) may alter the fatty acid composition of mitochondrial outer membranes and the characteristics of carnitine palmitoyltransferase I (CPTI) activity in the liver of normal Wistar rats. The animals were fed diets containing different oil mixtures (5% wt/wt) with the same ratio of n-6 n-3 fatty acids supplied either as fish oil or arachidonic acid concentrate. The cholesterol content of the mitochondrial outer membranes from liver was similar for all diets, while the percentage of 22:6n-3 and 20:4n-6 in phospholipids was enhanced with the diets containin…