Arachidonic acid maldistribution in obesity

Arachidonic acid is an important regulator of cellular function via its effects on the physical properties of membranes, in its free form, or as a substrate for eicosanoids. Dietary studies indicate that its production is regulated, but the mechanisms of this regulation and factors influencing arachidonate distribution from the site of production remain to be determined. In particular, whether there is a nonoxidative fate for arachidonate once it has been released from phospholipid has yet to be determined. Variations in the arachidonate content of serum, liver, and muscle lipid fractions have been correlated with alterations in lipogenesis and insulin action, implying a role for arachidonate in fuel partitioning. Evidence for this mechanism acting systemically has been found in genetic models of obesity in rodents and also in humans. This review proposes that variation in the distribution of arachidonate between phospholipid and cholesteryl ester fractions participates in the abnormal fuel partitioning associated with some forms of genetic obesity.     

Arachidonic acid and eicosapentaenoic acid stimulate type II pneumocyte surfactant secretion

Arachidonic acid and its leukotriene metabolites have been shown to stimulate surfactant secretion by alveolar type II cells. The present study was undertaken to determine the effects of various unsaturated fatty acids, including eicosapentaenoic acid, on surfactant secretion. Surfactant secretion was expressed as the percent of [³H]choline-derived phospholipids released into culture medium by type II pneumocytes of adult rats. Consistent with the earlier findings, arachidonic acid stimulated secretion in a concentration-dependent fashion (3.5–21 μM), doubling baseline secretion at 21 μM. Eicosapentaenoic acid was found to be equally effective as arachidonic acid in stimulating secretion. A comparison with palmitic, oleic and linoleic acids revealed that highly unsaturated fatty acids stimulated secretion to the greatest extent. This was supported by a positive correlation between degree of unsaturation (i.e., 0, 1, 2, 4 and 5 double bonds) and stimulation of surfactant secretion. In the present study, exogenous leukotriene E₄ (10⁻¹³–10⁻⁶) did not stimulate surfactant secretion. Neither nordihydroguaiaretic acid (0.1μM) nor indomethacin (0.1μM) affected arachidonic acid-stimulated secretion. The stimulatory effects of arachidonic acid and eicosapentaenoic acid on surfactant secretion were related to the highly unsaturated nature of the fatty acids and were not mediated by increased levels of cyclic adenosine monophosphate or calcium.     

The metabolism of polyunsaturated fatty acids in rat sertoli and germinal cells

The metabolism of [1-¹⁴C]linoleic, [1-¹⁴C]arachidonic and [3-¹⁴C]docosa-4,7,10,13,16-pentaenoic acids was investigated after intratesticular injection of the labeled compounds and isolation of rat Sertoli and germinal cells. Following injection of either¹⁴C-linoleate or¹⁴C-arachidonate, the specific activity (sp act) of docosa-4,7,10,13,16-pentaenoic acid of Sertoli cells was greater than that of the germinal cells. The data suggest that the Sertoli cells are more active in the biosynthesis of the 22-carbon pentaene than the germinal cells. Differences between these 2 cell types were also noted in the distribution of the incorporated¹⁴C among the various lipid classes. Following intratesticular injection of¹⁴C-docosapentaenoic acid, a greater proportion of the recovered¹⁴C in Sertoli cells than in germinal cells was present in 20-carbon fatty acids, suggesting a greater activity in Sertoli cells in the metabolism of the pentaene. The major portion of the recovered¹⁴C in both cell types was present in triacylglycerols during early time periods and in phospholipids after 24 hr. The possibility of transfer of biosynthesized docosapentaenoic acid from Sertoli to germinal cells is discussed.     

Pharmacological modulation of fatty acid desaturation and of cholesterol biosynthesis in THP-1 cells

In THP-1 cells, simvastatin decreases, in a concentration-dependent manner, cholesterol synthesis and increases linoleic acid (LA) conversion to its long-chain derivatives, in particular to arachidonic acid, activating Δ6 and Δ5 fatty acid (FA) desaturases. The intermediates in cholesterol synthesis, mevalonate and geranylgeraniol, partially reverse the effects of simvastatin on the LA conversion. The aims of this work were to evaluate: (i) the correlation between cholesterol synthesis and desaturase activity and (ii) the possible involvement of protein isoprenylation in desaturase activity, assessed through pharmacological treatments. THP-1 cells were incubated with [1-¹⁴C]LA or with [1-¹⁴C]di-homo-γ-linolenic acid (DHGLA) and treated with simvastatin or with curcumin and nicardipine, inhibitors of desaturases. Curcumin was more active than nicardipine in inhibiting LA and DHGLA conversion: 20 μM curcumin, alone or with simvastatin, totally inhibited Δ6 and Δ5 desaturation steps; 10 μM nicardipine only partially inhibited the enzymes, being more active on Δ5 desaturase. Simvastatin treatment decreased the incorporation of acetate in cholesterol (−93.8%) and cholesterol esters (−70.2%), as expected. Curcumin and nicardipine also decreased cholesterol synthesis and potentiated simvastatin. Finally, the isoprenylation inhibitors (perillic acid and GGTI-286) neither affected the conversion of LA nor inhibited the Δ5 desaturase activity. In conclusion, our results indicate that there is no direct relationship between cholesterol synthesis and desaturase activity. In fact, simvastatin decreased cholesterol synthesis and enhanced LA conversion (mainly Δ5 desaturation), whereas curcumin and nicardipin decreased Δ5 desaturation, with a limited effect on cholesterol synthesis.     

Arachidonic acid supply and metabolism in human infants born at full term

Infants need arachidonic acid (AA; C20:4n-6) for eicosanoid synthesis and deposition in growing tissues, including brain. Human milk supplies performed AA in amounts considered to meet accretion in membrane-rich tissues, but vegetable oil-based infant formulas do not contain AA. We studied two groups of ten healthy infants, each fed human milk or formula, and analyzed plasma lipid composition. Percentage contributions of AA to plasma phospholipids were stable over two months after birth in breast-fed infants, but infants fed formula developed significantly (P<0.05) lower levels at the ages of two weeks (formula 6.9% vs. breast 9.4%, w/w), one month (6.2 vs. 9.1%), and two months (5.7 vs 8.4%). In a second trial, we randomized infants to receive (from birth to age four months) formula without or with both AA and docosahexaenoic acid (DHA; C22:6n-3) at levels typical for mature human milk. Infants fed conventional formula showed a continuous decrease of phospholipid AA over time, whereas feeding of formula supplemented with AA and DHA led to significantly higher AA levels, similar to those in breast-fed infants (two months: supplemented 9.6% vs. unsupplemented 7.1%; four months; 8.7 vs. 6.6%). In order to estimate infantile capacity for endogenous synthesis of AA, we fed four term neonates with newly diagnosed phenylketonuria (mean age 18 d) a formula with all fat contributed by corn oil, which has a higher natural¹³C-enrichment than European human milk or formula. Analysis of¹³C-enrichment in plasma fatty acids over four days allowed us to estimate infantile AA synthesis. We found an increased¹³C-value in plasma AA of all infants, which indicates that term neonates can synthesize AA. However, with a simplified isotope balance equation, we estimate that endogenous synthesis contributed only about 23% of total plasma arachidonic acid by day four. We conclude that full-term infants fed formula may require a dietary supply of some preformed AA if the biochemical status of breast-fed infants is to be achieved. Based on a presentations at the AOCS Annual Meeting & Expo in San Antonio, Texas, May 7‐11, 1995.     

Regulation of fatty acid biosynthesis in Ehrlich cells by ascites tumor plasma lipoproteins

Fatty acid biosynthesis in Ehrlich cells in vitro was reduced when very low density lipoproteins (VLDL) isolated from the ascites tumor plasma were added to the incubation medium. The degree of inhibition was dependent on the VLDL concentration. At the VLDL concentrations usually present in the ascites plasma, there was a 30% decrease in biosynthesis as measured by³H₂O incorporation into fatty acids. Analysis of the labeled fatty acids by gas liquid chromatography indicated that this decrease was due to a reduction in fatty acid de novo biosynthesis and that chain elongation actually was increased when VLDL were present. Although ascites plasma low- and high density lipoproteins also produced a concentration-dependent inhibition of fatty acid biosynthesis, their effects were much smaller than those of the VLDL. Studies employing VLDL and radioactive free fatty acids indicated that the cells took up and utilized fatty acids derived from these lipoproteins. When VLDL were present, labeled free fatty acid incorporation into cell phospholipids, cholesteryl esters, and CO₂ decreased, whereas its incorporation into the cell free fatty acid pool increased. By contrast, the cells incorporated only very small amounts of fatty acid from either low- or high density lipoproteins. This suggests that the VLDL exert their inhibitory effect on fatty acid synthesis by supplying exogenous fatty acids to the cells. Presented in part at the AOCS Spring Meeting, Dallas, April 1975.     

Arachidonic acid to eicosapentaenoic acid ratio in blood correlates positively with clinical symptoms of depression

In this study of 20 moderately to severely depressed patients, diagnosed using current research diagnostic criteria and excluding known bipolar affective disorder and reactive depression, we investigated relationships between severity of depression and levels and ratios of n-3 and n-6 long-chain polyunsaturated fatty acids (PUFA) in plasma and erythrocyte phospholipids (PL). Severity of depression was measured using the 21-item Hamilton depression rating scale (HRS) and a second linear rating scale (LRS) of severity of depressive symptoms that omitted anxiety symptoms. There was a significant correlation between the ratio of erythrocyte PL arachidonic acid (AA) to eicosapentaenoic acid (EPA) and severity of depression as rated by the HRS (P<0.05) and the LRS for depression (P< 0.01). There was also a significant negative correlation between erythrocyte EPA and the LRS (P<0.05). The AA/EPA ratio in plasma PL and the ratio of erythrocyte long-chain (C₂₀ and C₂₂ carbon) n-6 to long-chain n-3 PUFA were also significantly correlated with the LRS (P<0.05). These findings do not appear to be simply explained by differences in dietary intake of EPA. We cannot determine whether the high ratios of AA/EPA in both plasma and erythrocyte PL are the result of depression or whether tissue PUFA change predate the depressive symptoms. We suggest, however, that our findings provide a basis for studying the effect of the nutritional supplementation of depressed subjects, aimed at reducing the AA/EPA ratio in tissues and severity of depression.     

Effect of various steroids on the biosynthesis of arachidonic acid in isolated hepatocytes and HTC cells

The effect of various steroids on the incorporation and desaturation of eicosa-8,11,14-trienoic acid in normal hepatocytes and HTC cells was investigated. After 3 hr incubation with 11-deoxycorticosterone, both kinds of cells showed an increase in the incorporation of eicosatrienoic acid. In contrast, progesterone, cortexolone, 17-β-estradiol, testosterone, estriol, aldosterone, corticosterone, dexamethasone, dehydroepiandrosterone, 11-β-hydroxyandrosterone, 11-ketoaetiocholanolone, epiaetiocholanolone and 5-β-pregnane-3α,20α-diol, provoked no significant changes in the uptake of the exogenous acid. Of all the steroids tested, only 11-dexycorticosterone, dexamethasone and 17-β-estradiol evoked a significant inhibition on the arachidonate biosynthesis in both kinds of cells. Testosterone, estriol, aldosterone and corticosterone provoked a significant inhibition of Δ5-desaturase in HTC cells. In dexamethasone, this effect was dose-dependent (0 to 10⁻⁴ M). Simultaneous incubation with 17-β- estradiol or 11-deoxycorticosterone with dexamethasone led to an extent of inhibition on arachidonate biosynthesis that did not surpass the effect of each drug. Pretreatment of isolated hepatocytes with the antiglucocorticoid, cortexolone, prevented the dexamethasone-induced inhibition of arachidonate biosynthesis. Normal rat liver microsomes preincubated in vitro with dexamethasone, 11-deoxycorticosterone, 17-β-estradiol, corticosterone or estriol (10⁻⁶ or 10⁻⁴ M concentration), showed no significant changes in the Δ5-desaturase activity. The results obtained suggest that the effect of the steroids on arachidonic acid biosynthesis in normal hepatocytes and HTC cells requires receptor occupancy and probably is mediated through a common biochemical mechanism.     

Arachidonic acid uptake by phospholipids and triacylglycerols of rat brain subcellular membranes

In the presence of ATP, MgCl₂ and CoASH, different subcellular membrane fractions isolated from rat cerebral cortex exhibited characteristic profiles for the incorporation of [1-¹⁴C]arachidonic acid into phospholipids and triacylglycerols. In general, uptake of label by phosphatidylcholines was higher in the synaptic membranes, and that by phosphatidylinositols was higher in the microsomes and somal plasma membranes. A substantial amount of the labeled arachidonate was also incorporated into triacylglycerols, especially in the somal plasma membranes and microsomes. Enzymes mediating the transfer of arachidonic acid to phospholipids were unstable with respect to sample storage and exposure to elevated temperatures. In contrast, the acyltransferase for triacylglycerols was more stable to these factors. Washing the membranes with bovine serum albumin resulted in an enhancement of the incorporation of label into phosphatidylinositols without affecting that of phosphatidylcholines, but the incorporation into triacylglycerols was inhibited. Treatment of synaptosomes and plasma membranes with saponin resulted in an enhancement in the labeling of phospholipids, but the labeling of triacylglycerols was inhibited. Thus, although labeled arachidonic acid was incorporated into phospholipids and triacylglycerols in brain subcellular membranes, these two types of acyltransferases exhibited different properties and responded differently to membrane perturbing agents.     

重组大肠杆菌全细胞用于苯乙酮酸的绿色生物合成

苯乙酮酸是化学合成中重要的合成砌块,可用于合成多种药物中间体,探索苯乙酮酸的绿色合成工艺具有重要的意义。以包含D-扁桃酸脱氢酶LhDMDH编码基因的重组大肠杆菌全细胞为催化剂,考察了它在无辅酶和辅底物添加的条件下对D-扁桃酸生物转化的效果,并对催化产物进行了纯化和鉴定。结果表明,本研究成功实现了在无辅酶和辅底物添加条件下苯乙酮酸的生物合成,产物的得率和纯度分别为45%和99%左右。成果也为外消旋扁桃酸的手性拆分及苯乙酮酸的生物合成奠定了基础。     

A comparative study of the lipid composition of isolated rat sertoli and germinal cells

The lipid composition of enriched preparations of sertoli cells and of germinal cells, isolated from the testes of mature rats, has been investigated. Sertoli cells contained a much lower content of phospholipids (in particular, much less phosphatidylcholine and phosphatidylethanolamine) and a higher content of triacylglycerols than did germinal cells. In addition, the Sertoli cells had a higher ratio of esterified to unesterified cholesterol than did germinal cells. Total lipids of Sertoli cells contained considerably lower levels of palmitic and docosa-4,7,10,13,16-pentaenoic acids and higher levels of stearic and oleic acids than did the total lipids of germinal less palmitic and docosa-4,7,10,13,16-pentaenoic acids, more stearic and oleic acids and also more arachidonic acid than did the corresponding lipid classes of the germinal cells. Minor differences between cell types were also noted for the content of palmitoleic, linoleic, docosa-7,10,13,16-tetraenoic, docosa-4,7,10,13,16,19-hexaenoic and tetracosa-9,12,15,18-tetraenoic acids.     

Arachidonic acid autoxidation in an aqueous media effect of α-tocopherol,cysteine and nucleic acids

The autoxidation of arachidonic acid dispersed in aqueous media was evaluated simultaneously with and without different agents, e.g., α-tocopherol at different concentrations, cysteine, DNA and RNA. The autoxidation rate of arachidonic acid was evaluated by quantitative gas liquid chromatography (GLC) determination of the unoxidized acid and by spectrophotometric measurement of conjugated dienes. α-Tocopherol exhibited a prooxidant activity at concentrations of 1.25 × 10⁻⁴ M and 1.25 × 10⁻⁵ M and a weak antioxidant activity at a concentration of 1.25 × 10⁻⁶ M. Cysteine showed antioxidant activity and greatly reduced the prooxidant activity of α-tocopherol. DNA and RNA had no effect in either case. α-Tocopherol oxidation was followed by high pressure liquid chromatography (HPLC). The prooxidant effect was accompanied by a rapid oxidation of α-tocopherol, except in the presence of cysteine, which prevented the oxidation of α-tocopherol.     

生物法合成5-氨基乙酰丙酸的研究进展

对国内外生物法合成5-氨基乙酰丙酸(ALA)的进展进行综述,重点比较了诱变育种法和代谢工程技术在ALA生物合成研究中的应用,揭示了新兴的代谢工程技术在ALA的生产开发中极具应用前景,为生物法合成ALA的进一步发展指明了方向,有利于促进ALA生物合成的产业化.     

Lipid and fatty acid biosynthesis by Rhodotorula minuta

Demand for fatty acids is increasing at an annual rate of 17%, due to their increased use in the oleochemical and transport industries. Presently, vegetable oils are the major source of fatty acids, whereas lipids with fatty acids similar to those of some vegetable oils have been reported to be synthesized by oleaginous microorganisms. In the present study, the culturing conditions for the oleaginous yeast Rhodotorula minuta IIP-33 have been optimized. In contrast to the lipid accumulation characteristics of most oleaginous yeasts, a carbon-to-nitrogen ratio of 30 was favorable for maximal accumulation of lipids (48%) with 22.5% conversion of glucose as carbon substrate. The lipids contained fatty acids in the C₇–C₁₈ range, the relative composition of which varied with culture temperature.     

Fatty acid biosynthesis in the developing endosperm ofCocos nucifera

Endosperm tissue of developing coconut endosperm incorporated [¹⁴C] acetate and [¹⁴C]-malonate into [¹⁴C]C₈-C₁₈ fatty acids. The distribution of [¹⁴C] label into the various fatty acids mimicked the distribution of endogenous fatty acids at early and middle stages of endosperm development. Although [¹⁴C] C₈-C₁₈ fatty acids were taken up rapidly by endosperm tissue slices, no elongation occurred; [¹⁴C] stearic acid was not desaturated to oleic. Cell free preparations have also been obtained from this tissue that readily incorporated [¹⁴C] malonyl-CoA into a range of [¹⁴C] fatty acids in the presence of ACP and NADH at pH 7.0. Employing this system, a number of experiments were designed to determine the mechanism of chain length termination. In contrast to intact tissue slice experiments, cell-free extracts yielded as principal products palmitic and stearic acid, although up to 20% were shorter chain acids. A number of possible mechanisms for chain length termination were proposed and tested. Supported in part by NSF Grant No. PCM76 01495.     

Arachidonic acid intestinal absorption: Mechanism of transport and influence of luminal factors on absorption in vitro

The mechanism and characteristics of intestinal absorption of arachidonic acid were studied in vitro using everted intestinal sacs of the rat. Arachidonic acid absorption was studied at concentrations of 5 μM to 8.36 mM. The plot of absorption rate vs. concentration fitted best to a rectangular hyperbola at low μM concentrations and to a straight linear relationship in the mM range of concentrations. Metabolic inhibitors and uncouplers did not change absorption in either range of concentrations. The absorption of arachidonic acid increased with thinning of the unstirred water-layer, decrease in the pH, or the substitution of sodium taurocholate by Pluronic F 68 or Tween 80. Absorption decreased following the equimolar additions of oleic, linoleic, and linolenic acids. Absorption rate did not change when the taurocholate concentration was varied from 5–15 mM or following the additions of butyric or glutamic acids, leucine, lysine, or dextrose. It was concluded that arachidonic acid is absorbed by a concentration-dependent dual mechanism of transport which is not energy dependent. At the low μM range of concentrations, facilitated diffusion is predominant, while at mM concentrations, simple diffusion is the dominant mechanism of absorption. Changes in the intestinal fluid composition, flow rate, and pH can modify the rate of absorption of arachidonic acid.     

Arachidonic acid supplementation enhances synthesis of eicosanoids without suppressing immune functions in young healthy men

This study was conducted to determine the effects of arachidonic acid (AA) supplementation on human immune response (IR) and on the secretion of prostaglandin E2 (PGE2) and leukotriene B4 (LTB4). Ten healthy men (20–38 yr) participated in the study and lived at the Metabolic Suite of the Western Human Nutrition Research Center. They were fed a basal diet (57, 27, and 16 energy percentage from carbohydrate, fat, and protein, respectively, and AA 200 mg/d) for the first 15 d of the study. Additional AA (1.5 g/d) was added to the diet of six men from day 16 to 65, while the remaining four subjects remained on the basal diet. The diets of the two groups were crossed-over from day 66 to 115. In vitro indices of IR were examined using blood drawn on days 15, 58, 65, 108, and 115. Influenza antibody titers were determined in the sera prepared from blood drawn on days 92 and 115 (23 d postimmunization). AA supplementation caused significant increases in the in vitro secretion of LTB4, and PGE2, but it did not alter the in vitro secretion of tumor necrosis factor α; interleukins 1β, 2, 6; and the receptor for interleukin 2. Nor did it change the number of circulting lymphocytes bearing markers for specific subsets (B, T, helper, suppressor, natural killer) and the serum antibody titers against influenza vaccine. The opposing effects of PGE2 and LTB4 may have led to the lack of change in immune functions tested.