Dietary supplementation with docosahexaenoic acid, but not eicosapentaenoic acid, dramatically alters cardiac mitochondrial phospholipid fatty acid composition and prevents permeability transition

Treatment with the ω-3 polyunsaturated fatty acids (PUFAs) docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) exerts cardioprotective effects, and suppresses Ca²⁺-induced opening of the mitochondrial permeability transition pore (MPTP). These effects are associated with increased DHA and EPA, and lower arachidonic acid (ARA) in cardiac phospholipids. While clinical studies suggest the triglyceride lowering effects of DHA and EPA are equivalent, little is known about the independent effects of DHA and EPA on mitochondria function. We compared the effects of dietary supplementation with the ω-3 PUFAs DHA and EPA on cardiac mitochondrial phospholipid fatty acid composition and Ca²⁺-induced MPTP opening. Rats were fed a standard lab diet with either normal low levels of ω-3 PUFA, or DHA or EPA at 2.5% of energy intake for 8 weeks, and cardiac mitochondria were isolated and analyzed for Ca²⁺-induced MPTP opening and phospholipid fatty acyl composition. DHA supplementation increased both DHA and EPA and decreased ARA in mitochondrial phospholipid, and significantly delayed MPTP opening as assessed by increased Ca²⁺ retention capacity and decreased Ca²⁺-induced mitochondria swelling. EPA supplementation increased EPA in mitochondrial phospholipids, but did not affect DHA, only modestly lowered ARA, and did not affect MPTP opening. In summary, dietary supplementation with DHA but not EPA, profoundly altered mitochondrial phospholipid fatty acid composition and delayed Ca²⁺-induced MPTP opening.     

Characterization of the phospholipid and fatty acid composition of Sendai virus

The lipid composition of Sendai virus, propagated in chicken eggs, was analyzed by high performance liquid chromatography (HPLC), thin-layer chromatography (TLC), and gas-liquid chromatography (GLC). Phosphatidylcholine was found to be the dominant phospholipid (37.3%) with phosphatidylethanolamine (26.8%) and phosphatidylserine (12.0%) also present in significant amounts. Analysis of the fatty acid methyl esters revealed that the dominant fatty acids in total phospholipid were: C16:0 (17.6%), C18:0 (15.4%), C18:1 (n-9) (22.0%), and C24:0 (6.0%). Cardiolipin, phosphatidylserine, and sphingomyelin contained higher levels of saturated fatty acids relative to phosphatidylinositol, phosphatidylethanolamine, and phosphatidylcholine.     

Training affects muscle phospholipid fatty acid composition in humans.

Training improves insulin sensitivity, which in turn may affect performance by modulation of fuel availability. Insulin action, in turn, has been linked to specific patterns of muscle structural lipids in skeletal muscle. This study investigated whether regular exercise training exerts an effect on the muscle membrane phospholipid fatty acid composition in humans. Seven male subjects performed endurance training of the knee extensors of one leg for 4 wk. The other leg served as a control. Before, after 4 days, and after 4 wk, muscle biopsies were obtained from the vastus lateralis. After 4 wk, the phospholipid fatty acid contents of oleic acid 18:1(n-9) and docosahexaenoic acid 22:6(n-3) were significantly higher in the trained (10.9 +/- 0.5% and 3.2 +/- 0.4% of total fatty acids, respectively) than the untrained leg (8.8 +/- 0.5% and 2.6 +/- 0.4%, P < 0.05). The ratio between n-6 and n-3 fatty acids was significantly lower in the trained (11.1 +/- 0.9) than the untrained leg (13.1 +/- 1.2, P < 0.05). In contrast, training did not affect muscle triacylglycerol fatty acid composition. Citrate synthase activity was increased by 17% in the trained compared with the untrained leg (P < 0.05). In this model, diet plays a minimal role, as the influence of dietary intake is similar on both legs. Regular exercise training per se influences the phospholipid fatty acid composition of muscle membranes but has no effect on the composition of fatty acids stored in triacylglycerols within the muscle.     

Specific phospholipid fatty acid composition of brain regions in mice. Effects of n-3 polyunsaturated fatty acid deficiency and phospholipid supplementation

This study examined the effects of dietary alpha-linolenic acid deficiency followed or not by supplementation with phospholipids rich in n;-3 polyunsaturated fatty acid (PUFA) on the fatty acid composition of total phospholipids in 11 brain regions. Three weeks before mating, mice were fed a semisynthetic diet containing both linoleic and alpha-linolenic acid or deficient in alpha-linolenic acid. Pups were fed the same diet as their dams. At the age of 7 weeks, a part of the deficient group were supplemented with n;-3 polyunsaturated fatty acids (PUFA) from either egg yolk or pig brain phospholipids for 2 months. Saturated and monounsaturated fatty acid levels varied among brain regions and were not significantly affected by the diet. In control mice, the level of 22:6 n-3 was significantly higher in the frontal cortex compared to all regions. alpha-Linolenic acid deficiency decreased the level of 22:6 n-3 and was compensated by an increase in 22:5 n-6 in all regions. However, the brain regions were affected differently. After the pituitary gland, the frontal cortex, and the striatum were the most markedly affected with 40% reduction of 22:6 n-3. Supplementation with egg yolk or cerebral phospholipids in deficient mice restored a normal fatty acid composition in brain regions except for the frontal cortex. There was a regional distribution of the fatty acids in the brain and the impact of deficiency in alpha-linolenic acid was region-specific. Dietary egg yolk or cerebral phospholipids are an effective source of n-3 PUFA for the recovery of altered fatty acid composition induced by a diet deficient in n-3 PUFA.     

Effect of lidocaine on phospholipid and fatty acid composition of bacterial membranes

The effects of lidocaine on chemical composition of membrane phospholipids and membrane fluidity of Streptococcus mutans have been studied. Increasing concentra-tions of lidocaine induced both an increase in cardiolipin and a decrease in the degree of unsaturation of its fatty acid composition. A lidocaine-dependent decrease of membrane fluidity was observed from an electron spin resonance spectroscopic study. It was considered thal bacteria grown with lidocaine below its minimum inhibitory concentration resisted the effect of the drug by modifying phospholipid and fatty acid composition resulting in a decreased membrane fluidity.     

Lipid composition, phospholipid profile and fatty acid of rat caecal mucosa.

In this study, we examined the lipid composition of rat caecal mucosa, including the fatty acid composition of major phospholipid classes. Phospholipids accounted for 90% of the total lipid, with cholesterol, triacylglycerols, diacylglycerols, fatty acids and cholesterol ester making up the remainder. Therefore, a phospholipid to neutral lipid ration of 9:1 was found. Phosphatidylethanolamine was the predominant phospholipid, with phosphatidylcholine as the second most abundant phospholipid. Cardiolipin, phosphatidylserine, phosphatidylinositol and lysophosphatidylcholine were present in lesser amounts. Sphingomyelin and lysophosphatidylethanolamine were only detected in trace amounts. The major fatty acids present in both the lipid and all phospholipid fractions were palmitate, stearate, oleate, linoleate and arachidonate. Other fatty acids of chain length greater than C20 were only detected in phospholipid fraction and accounted for < 5% of the total fatty acids in this fraction. However, 11.10% of 22:6 (n-3) and 7.17% of 24:0 were detected in phosphatidylserine and lysophosphatidylcholine, respectively. The results are discussed in terms of their possible physiological significance.     

Relationships between heat resistance and phospholipid fatty acid composition of Vibrio parahaemolyticus.

Vibrio parahaemolyticus was grown in tryptic soy broth (TSB) containing NaCl levels of 0.5, 3.0, and 7.5% (wt/vol). Cultures incubated at 21, 29, and 37 C were harvested in late exponential phases and thermal death times at 47 C (D47 c; time at 47 C required to reduce the viable population by 90%) were determined in phosphate buffer containing 0.5, 3.0, and 7.5% NaCl. At a given NaCl concentration in the growth medium, D47 c values increased with elevated incubation temperatures and with elevated levels of NaCl in the heating menstrua. Differences in thermal resistance of cells cultured at a particular temperature were greater between those grown in TSB containing 0.5 and 3.0% NaCl than between those grown in TSB containing 3.0 and 7.5% NaCl. D47c values ranged from 0.8 min (grown at 21 C in TSB with 0.5% NaCl) to 6.5 min (grown at 37 C in TSB with 7.5%, heated in 7.5% NaCl buffer). Methyl esters of major phospholipid fatty acids extracted from cells were quantitated. The ratio of saturated to unsaturated fatty acids in cells grown at a given NaCl concentration increased with elevated incubation temperature. At a particular growth temperature, however, saturated to unsaturated fatty acids ratios were lowest for cells grown in TSB containing 3.0% NaCl.     

磷脂脂肪酸法在土壤微生物群落分析中的应用

土壤微生物群落的组成一直是土壤学、微生物学和生态学研究的热点问题。我国在这方面的研究处于国际前列,越来越多的研究成果在国际重要刊物上发表。而磷脂脂肪酸(PLFA)法在土壤微生物群落分析中占有举足轻重的地位,国内外学者都热衷于使用该方法。但是PLFA法的使用仍存在一些不足的地方,需要研究学者们慎重使用。本文综述了国际上相关研究,概述了PLFA方法使用的发展历史,应用及挑战。总结了使用和数据解读时需要注意的问题,整理了PLFA法相关的生物标记以及与新方法结合的设想,方便以后研究的开展。     

Comparison of phospholipid and fatty acid composition of wild and cultured striped bass eggs

The dominant fatty acids in all neutral lipid fractions of non-water hardened eggs from two wild and one cultured stock of striped bass Morone saxatilis were the monoenes, 18 : 1n9/n7>16 : 1n7>17 : 1. The dominant fatty acids in the phospholipid fraction of all eggs, regardless of origin, were 22 : 6n3>18 : 1n9/n7>20 : 5n3>16 : 1n7>16 : 0>18 : 0. Arachadonic acid (AA, 20 : 4n6) was significantly lower (2·0%) in cultured fish eggs compared to either wild stock (5·8–6·1%). Fatty acids from the liver and eggs of wild Shubenacadie fish were similar to one another with respect to both neutral and phospholipid fractions. However, the AA and eicosapentaenoic acid (EPA, 20 : 5n3) content of the phospholipid fraction varied according to the hypothesized migration behaviour of Shubenacadie fish. The total lipid content of wild fish eggs was significantly greater than that of cultured fish. The total phospholipid content of Shubenacadie eggs was significantly higher than either Roanoke or cultured fish eggs. Phosphotidylinositol (PI) was the dominant phospholipid found ins all egg samples from all origins as opposed to phosphotidylcholine, which is usually the dominant phospholipid. These data indicate that PI and AA may have important and as yet unidentified roles in fertilization and embryonic development in these fish.     

Effects of pesticides on the fatty acid and phospholipid composition of Escherichia coli.

Cells of Escherichia coli contained an altered phospholipid and fatty acid composition when grown in the presence of some pesticides. Whereas parathion increased the concentration of all phospholipid species without changes in their polar head groups. DDT (dichlorodiphenyltrichloroethane) decreased the proportion of neutral serine-derived phosphatides and dieldrin decreased the proportion of negatively charged phospholipids. The saturated/unsaturated plus cyclopropane fatty acid ratio was increased in all cases. The changes suggested that cells adapted their membrane lipids to compensate for the presence of pesticides in the environment.     

Brain and hippocampus fatty acid composition in phospholipid classes of aged-relative cognitive deficit rats

The aim of this work was to study the composition of long chain fatty acids and the n-6 and n-3 fatty acid ratios in aged and young Wistar rats in brain and hippocampus, related to relative cognitive deficits. The aged animals showed cognitive deficits during acquisition of a memory task (delayed alternation). In brain, results showed a decrease in palmitoleic and palmitic acid percentages in all the studied phospholipid classes and in the phosphatidylserine and phosphatidylcholine classes, respectively, in old rats, compared to the young ones. There was also an increase in oleic and stearic acid amounts in the sphingomyelin, phosphatidylserine and phosphatidylinositol classes and in the phosphatidylserine and phosphatidylcholine classes, respectively. Arachidonic acid amount was decreased in old rats, compared to the young ones, in the phosphatidylserine and phosphatidylinositol classes. Total n-6 and n-3 fatty acid amounts were both decreased in all phospholipid classes, with a stable n-6/n-3 ratio. Our results confirm that arachidonic acid concentration is decreased in aged rats and that this reduction, more significant in phosphatidylserine and phosphatidylinositol classes, should be related to the fact that low concentrations of arachidonic acid are observed during activation of glutamate receptor.     

Modification of the fatty acid composition of phospholipid in measles virus-persistently infected cells

The composition of phospholipid was studied in BGM cells uninfected, persistently infected or lytically infected with measles virus, strain Hallé. In persistently infected cells, phosphatidylcholine palmitic acid content, and phosphatidylethanolamine palmitic acid and arachidonic acid contents were significantly increased. Lytically infected cells had a similar phospholipid fatty acid composition to the uninfected. Phosphatide composition showed minor modifications, but the content of total choline-derivative phospholipids was unchanged in either type of infection.     

Effect of exogenous fatty acids on growth,membrane fluidity,and phospholipid fatty acid composition in yeast

The growth response of a double-mutant fatty acid auxotroph of yeast Saccharomyces cerevisiae to exogenous saturated fatty acids of a homologous series from 12:0 to 16:0, each supplied with oleate, linoleate, linolenate, or cis-Δ¹¹- eicosenoate, cannot be explained in terms of the efficiency of incorporation of the fatty acids into phospholipids or alteration of membrane fluidity. There is, however, a negative correlation between growth and levels of 12:0 plus 13:0 in phospholipids, as well as a positive correlation between growth and levels of 14:0, 1 5:0, and 1 6:0. We, therefore, conclude that the predominant factor in these phospholipid fatty acyl chain modifications is maintenance of an optimal concentration of C14:0 through C16:0 in phospholipids of this organism.     

Temperature-dependent changes in phospholipid and fatty acid composition and membrane lipid fluidity of Yersinia enterocolitica

Temperature-dependent compositional changes of phospholipids and their fatty acids were analysed in Yersinia enterocolitica grown at 5°, 25° and 37°C. The relative amounts of the four phospholipids, phosphatidylethanolamine (75–78%), phosphatidylglycerol (10–11%), cardiolipin (<7%) and lysophosphatidylethanolamine (<5%), were essentially the same at all growth temperatures. The degree of fatty acid unsaturation of the four phospholipids increased with decrease in growth temperature, mainly due to an increase of C_16:1 and C_18:1 and a corresponding decrease of C_16;0, C_18:0 and cyclo C_17:0. An electron spin resonance spectroscopic study of the membrane lipids showed that membrane lipid fluidity was enhanced by decreasing the growth temperatures. The changes in fatty acid composition of phospholipids in response to varied temperatures were consistent with the temperature-dependent changes in the membrane lipid fluidity of Y. enterocolitica , and were similar to those reported for other bacteria.     

Influence of N-palmitoylethanolamine on the phospholipid and fatty acid composition of rat liver in ischemia

The effect of N-palmitoylethanolamine (NPE, 10(-5) M) on the lipid peroxidation (LPO) processes, phospholipid and fatty acid content in the rat liver at perfusion and ischemia during the liver preservation was estimated. As early as at the 5th min of perfusion by cooled conserving solution "Eurocollins", LPO activation was determined. Simultaneously the content of lysophosphatidylcholine (LPC) and phosphatidylserine increased, the total cholesterol level decreased. The redistribution of saturated and unsaturated fatty acids quantity was detected. The addition of NPE into "Eurocollins" solution reduced the accumulation of malonedialdehyde and LPC, modified the fatty acids content. These effects, evidently, formed the basis of the protective action of NPE on the hepatic tissues under anoxia.     

Effect of phospholipid fatty acid composition of endothelial cells on cholesterol efflux rates.

Human endothelial cells (EA.hy 926 line) were loaded with cationized low density lipoprotein (LDL) and subsequently incubated with fatty acid/bovine serum albumin complexes. The fatty acids were palmitic, oleic, linoleic, arachidonic, and eicosapentaenoic acids. The preincubations resulted in extensively modified fatty acid profiles in cell membrane phospholipids and in cellular cholesteryl esters. The cholesterol efflux from these fatty acid-modified cells was measured using 0.2 mg high density lipoprotein3 (HDL3)/ml medium. The efflux was significantly higher for the palmitic acid-treated cells, compared to all other fatty acid treatments. These differences in efflux rates were not caused by changes in the binding of HDL3 to high affinity receptors on the EA.hy 926 cells. Efflux mediated by dimethyl suberimidate-treated HDL3, which does not interact with high affinity HDL receptors, was similar to efflux induced by native HDL3 after all fatty acid treatments. Our results indicate that high affinity HDL receptors are not important for HDL-mediated efflux of cell cholesterol. The fatty acid composition of the cell membrane phospholipids may be an important determinant.     

Effect of membrane fluidity and fatty acid composition on the prothrombin-converting activity of phospholipid vesicles.

Vesicles composed of phospholipids with different fatty acyl side chains have been utilized to examine the importance of the nonpolar membrane region for the prothrombin-converting activity of procoagulant phospholipid vesicles. Membranes composed of phosphatidylserine (PS) and phosphatidylcholine (PC) with unsaturated fatty acyl side chains were more active in prothrombin activation than membranes composed of phospholipids with saturated fatty acyl chains. This phenomenon was observed above the phase transition temperature, i.e., on membranes in the liquid-crystalline state. The prothrombin-converting activity of saturated phospholipids approached the activity of unsaturated phospholipids at high factor Va concentrations, which is indicative for a less favorable equilibrium constant for prothrombinase assembly on membrane surfaces composed of saturated phospholipids. The difference between saturated and unsaturated phospholipids was annulled on membranes with high mole percentages of PS. This may result from a compensating contribution of electrostatic forces to the binding equilibria involved in prothrombinase assembly. Additional effects on the prothrombin-converting activity were observed when membranes containing saturated phospholipids were studied below their phase transition temperature. In agreement with Higgins et al. [(1985) J. Biol. Chem. 260, 3604-3612], we found that the time required for the assembly of prothrombinase from membrane-bound factors Xa and Va is considerably prolonged on solid membranes. However, we also observed an effect of membrane fluidity on the steady-state rate of prothrombin activation. Kinetic experiments at saturating factor Va concentrations showed that the transition from the liquid-crystalline to the gel state caused a more than 9-fold decrease of the kcat of prothrombin activation without affecting the Km for prothrombin.(ABSTRACT TRUNCATED AT 250 WORDS)