Recent biochemical nutrition knowledge in relation to metabolism and the significance of essential fatty acids and n-3-fatty acids contained in fish

A survey is given on some newer knowledge about metabolism and about the importance of the essential fatty acids and of the n-3 fatty acids (eicosapentaenic, docosapentaenic, docosahexaenic acids) which occur in fish oils. In the body the linoleic acid via intermediate steps can be transformed into the arachidonic acid, from which various prostaglandins and leucotriens as well as the thromboxane A2 can be formed. The transformation of the linolenic acid into the eicosapentaenic acid is slight in man. The docosahexaenic acid is necessary for the construction of phospholipids in the brain and in the retina. The uptake of fish fatty acids inhibits the formation of thromboxane A2 and of leukotriens from the arachidonic acid. The fish fatty acids further in the liver in the peroxisomas the activity of the enzymes for the beta-oxidation; the formation of lipoproteins of high density increases under their influence: the triacylglyceride content, the cholesterol as well as the lipoprotein content of very low and low density decreases, when there is an adequate part of fish fatty acids in the nutrition.     

Cardiovascular disease-related genes and regulation by diet

Diets rich in omega-3 polyunsaturated fatty acids (n-3 PUFAs) such as α-linolenic acid, eicosapentaenoic acid, and docosahexaenoic acid are associated with decreased incidence and severity of cardiovascular disease (CVD). At least some of the beneficial effects of these dietary fatty acids are mediated by metabolites such as prostaglandins, leukotrienes, thromboxanes, and resolvins. The effects of n-3 PUFAs often differ from those of other fatty acids with very similar structures, such as linoleic acid and arachidonic acid (n-6 PUFAs) and their corresponding metabolites. This article reviews the evidence that specific receptors exist for fatty acids or their metabolites that are able to regulate gene expression and coordinately affect metabolic or signaling pathways associated with CVD. Four nuclear receptor subfamilies that respond to dietary and endogenous ligands and have implications for CVD are emphasized in this article: peroxisome proliferator-activated receptors, retinoid X receptors, liver X receptors, and the farnesoid X receptor.     

Development of alcoholic fatty liver and fibrosis in rhesus monkeys fed a low n-3 fatty acid diet

BACKGROUND: The amount and type of dietary fat seem to be important factors that modulate the development of alcohol-induced liver steatosis and fibrosis. Various alcohol-feeding studies in animals have been used to model some of the symptoms that occur in liver disease in humans. METHODS: Rhesus monkeys (Macaca mulatta) were maintained on a diet that had a very low concentration of alpha-linolenic acid and were given free access to an artificially sweetened 7% ethanol solution. Control and ethanol-consuming animals were maintained on a diet in which the linoleate content was adequate (1.4% of energy); however, alpha-linoleate represented only 0.08% of energy. Liver specimens were obtained, and the fatty acid composition of the liver phospholipids, cholesterol esters, and triglycerides of the two groups were compared at 5 years and histopathology of tissue samples were compared at 3 and 5 years. RESULTS: The mean consumption of ethanol for this group over a 5-year period was 2.4 g.kg.day. As a consequence of the ethanol-dietary treatment, there were significantly lower concentrations of several polyunsaturated fatty acids in the liver phospholipids of the alcohol-treated group, including arachidonic acid and most of the n-3 fatty acids and particularly docosahexaenoic acid, when compared with dietary controls. Liver specimens from animals in the ethanol group at 5 years showed a marked degree of steatosis, both focal and diffuse cellular necrosis, and an increase in the development of fibrosis compared with specimens obtained at 3 years and with those from dietary controls, in which there was no evidence of fibrotic lesions. CONCLUSION: These findings suggest that the advancement of ethanol-induced liver disease in rhesus monkeys may be modulated by the amount and type of dietary essential fatty acids and that a marginal intake of n-3 fatty acids may be a permissive factor in the development of liver disease in primates.     

Prostaglandin synthesis in goldfish heart, Carassius auratus

Potential precursors for prostaglandin (PG) synthesis were measured in goldfish heart and skeletal muscle by gas chromatography. Heart tissue contained docosahexaenoic, arachidonic, eicosapentaenoic, and eicosatrienoic acids in concentrations of 3223 +/- 128, 1216 +/- 7.8, 260 +/- 72.8, and 250 +/- 14 ng/mg wet wt, respectively. 14C-Labeled substrates were examined for their ability to be converted to prostaglandins. Eicosatrienoic and docosahexaenoic acid were not synthesized into prostaglandins, with 66 and 72% of the substrate remaining as free fatty acids, respectively. In contrast, both arachidonic and eicosapentaenoic acids were converted predominantly to PGFs and PGIs. The conversion was time dependent and complete by 30 min. The conversion patterns with eicosapentaenoic acid and arachidonic acid were essentially the same. The data suggest that goldfish cyclooxygenase can utilize two of the four potential substrates for prostaglandin synthesis. As fatty acid levels in fish vary with environmental temperature, substrate availability rather than cyclooxygenase preference may dictate the types of prostaglandins which are produced.     

Arachidonic acid is responsible for the smooth muscle-contracting activity of G-acid.

G-acid, a smooth-muscle-contracting substance isolated from human blood plasma, was previously identified as 3-octadecenoic acid. Synthetic cis- and trans-3-octadecenoic acid, however, are biologically inactive. G-acid actually consists of a mixture of fatty acids, characteristic of animal tissues. The biological activity is caused by arachidonic acid, which contracts smooth muscle upon in situ conversion to prostaglandins.     

Influence of different dietary fatty acid sources on erythrocyte lipids and plasma and liver essential fatty acids in hamsters fed ethanol

Hamsters fed ethanol were given three different dietary sources of essential fatty acids; safflower oil, evening primrose oil (both mainly n-6 fatty acids) or linseed oil (mainly n-3 fatty acids). After 7 weeks, plasma, erythrocyte and liver lipids and fatty acids were analyzed. Plasma and liver lipids were not significantly different in the ethanol-fed hamsters compared to the controls. Erythrocyte total phospholipid was increased only in the ethanol-fed groups given n-6 but not n-3 fatty acids. Some fatty acid changes induced by ethanol were predictable, e.g. lower 20:4 n-6 in hamsters fed n-6 fatty acids, but others were not predictable, e.g. higher 22:6 n-3 in all the ethanol-fed groups. The effect of ethanol on hamster lipids and fatty acid composition appears dependent on the predominant class of dietary fatty acids.     

Platelet function, thromboxane formation and blood pressure control during supplementation of the Western diet with cod liver oil

Epidemiologic and experimental data suggest an antiatherothrombotic potential of omega-3 polyunsaturated fatty acids. Therefore, the Western diet, which supplies predominantly omega-6 polyunsaturated fatty acids, was supplemented with 40 ml/day of cod liver oil, which provides about 10 g of omega-3 polyunsaturated fatty acids daily, for 25 days in eight volunteers. The omega-3 polyunsaturated fatty acids were incorporated in platelet and erythrocyte membrane phospholipids at the expense of omega-6 polyunsaturated fatty acids. Bleeding time increased (p less than 0.01) and platelet count (p less than 0.05), platelet aggregation upon ADP and collagen (p less than 0.01-0.05), and associated thromboxane B2 formation (p less than 0.01) decreased. Blood pressure (p less than 0.05) and blood pressure response to norepinephrine (p less than 0.01) and angiotensin II (NS) fell, without major changes in plasma catecholamines, renin, urinary aldosterone, kallikrein, prostaglandins E2 and F2 alpha and red cell cation fluxes. Biochemical and functional changes were reversed 4 weeks after cod liver oil was discontinued. Formation of prostaglandins derived from eicosapentaenoic acid and interference of eicosapentaenoic acid with formation and action of prostaglandins derived from arachidonic acid were evident in vitro. Whatever the mechanism, this moderate supplement of omega-3 polyunsaturated fatty acids markedly changed membrane phospholipids, which was associated with a shift toward less reactive platelets and a blunted circulatory response to pressure hormones.     

Long-term ethanol consumption in the hamster: effects on tissue lipids, fatty acids and erythrocyte hemolysis

Male Golden Syrian hamsters at 1 year of age were given a basal diet and either distilled water or 10% absolute ethanol in distilled water to drink for 1 year in order to determine the influence of prolonged ethanol intake on tissue long chain fatty acid, lipid composition and erythrocyte hemolysis in response to osmotic stress. Total lipids were extracted from liver, heart, plasma and erythrocytes. Individual lipid fractions were quantitated and the percentage fatty acid composition of the lipid fractions analyzed by gas-liquid chromatography. Although no significant changes in tissue lipid content or erythrocyte hemolysis were attributable to ethanol intake, fatty acid changes were marked in the ethanol-fed hamsters. The primary fatty acid changes were increased oleic acid (40-50%) and decreased linoleic acid (25-60%) which were observed in all tissues. Arachidonic acid was decreased only in triacylglycerol fractions. The results suggest that in the hamster long-term voluntary ethanol intake alters specific long chain fatty acids, but that erythrocyte membrane integrity and tissue lipid composition were not compromised.     

Alterations in the lipids and prostaglandins in mouse spleen following the ingestion of menhaden oil

Two groups of male mice were fed for 2 weeks with a semisynthetic diet supplemented with either 10% hydrogenated coconut oil or 10% menhaden oil. The spleen from animals fed with menhaden oil contained significantly higher amounts of polyunsaturated n-3 fatty acids. The n-3 fatty acids reciprocally replaced arachidonic acid in the phospholipids. The synthesis of 6-keto prostaglandin F1 alpha and prostaglandin E2 by spleen tissues were significantly depressed (70-80%) in mice consuming menhaden oil. These studies indicated that n-3 fatty acids can effectively displace arachidonic acid from spleen lipids and thereby affect the synthesis of prostaglandins. The implications of these observations are discussed.     

实验性脂肪肝肝纤维化组织中弹性蛋白表达的研究

目的探讨弹性蛋白表达与脂肪肝肝纤维化的关系。方法分别建立高脂饮食、低脂酒精饮食、高脂酒精饮食和四氯化碳大鼠实验性脂肪肝肝纤维化模型,用免疫组织化学方法观察造模肝组织中弹性蛋白的表达。结果低脂饮食组、高脂饮食组和低脂饮食酒精组未见弹性蛋白的表达;高脂饮食酒精组有部分表达,而四氯化碳组均见弹性蛋白的表达;窦周肝细胞表达强度明显高于肝细胞。结论在肝纤维化的发展过程中,弹性蛋白的表达见于肝纤维化后期,与肝纤维化程度相关,可反映肝纤维化的病程;窦周肝细胞（主要是肝星状细胞）在肝纤维化进程中起十分重要的作用。     

High-level expression of rat class I alcohol dehydrogenase is sufficient for ethanol-induced fat accumulation in transduced HeLa cells

The mechanisms by which ethanol causes fatty liver are complex. Reducing equivalents generated during ethanol oxidation inhibit tricarboxylic acid cycle activity and fatty acid oxidation. In addition, ethanol inhibits lipoprotein export and increases fatty acid uptake and lipid peroxidation. To test the role that alcohol metabolism by alcohol dehydrogenase (ADH) has on cellular lipid metabolism, a cell line expressing rat ADH was generated by transducing HeLa cells with an ADH-expressing retrovirus. The cells expressed high levels of ADH protein and had ADH activity similar to that of liver. Exposure of the cells to 20 mmol/L ethanol for 24 hours led to substantial accumulation of free fatty acids and triacylglycerol in the transduced, but not wild-type, HeLa cells. The rate of synthesis of saponifiable lipid was increased significantly by ethanol under these conditions. Ethanol exposure also promoted triacylglycerol accumulation when the cells were incubated with linoleic acid. This was associated with a decrease in the rate at which the cells oxidized 1-[14-C]-linoleic acid. Fat accumulation was not prevented by including alpha-tocopherol in the medium, arguing against a role for lipid peroxidation. However, the presence of methylene blue completely prevented the fat accumulation. This was associated with a return of the elevated lactate/pyruvate ratio toward normal. These data suggest that generation of reducing equivalents by ADH was sufficient to cause fat accumulation in this cell model.     

Vitamin E intake affects serum thromboxane and tissue essential fatty acid composition in the rat

The influence of dietary vitamin E on the composition of essential fatty acids in rat tissue and plasma lipids as well as serum thromboxane B2 was studied. Diets containing deficient (0 mg/kg diet), adequate (100 mg/kg) or supplemental (1,000 mg/kg) vitamin E were fed to young male rats for 10 weeks. The ratio of dihomo-gamma-linolenic acid to arachidonic acid in phospholipids of plasma, liver, and testes was increased in vitamin-E-supplemented rats. Serum thromboxane B2 was increased in vitamin-E-deficient rats. The data support a role for vitamin E in affecting both metabolism of long chain fatty acids, i.e. dihomo-gamma-linolenic acid, and conversion of arachidonic acid to thromboxane A2.     

Effect of ethanol on liver triglycerides and fatty acid composition in the golden Syrian hamster

In choice situations, the golden Syrian hamster shows a strong preference to drink an ethanol solution to water alone. Because of the known effects of ethanol on lipid metabolism, its effects on liver fatty acids were therefore studied in the hamster. After a period of 8 weeks during which hamsters had consumed water containing 10% ethanol (4 weeks) and then 15% ethanol (4 weeks), the liver was significantly increased in weight, an effect which was shown to be due to fat accumulation as triglyceride. The fatty acid composition of the liver triglycerides and phospholipids was altered such that monounsaturated fatty acids were very significantly increased and both n-6 and n-3 essential fatty acids were proportionally decreased.     

Contrasting effects of low or high copper intake on rat tissue lipid essential fatty acid composition

The effects of low copper intake or copper supplementation on the metabolism of stearic acid have been studied previously, but their effects on essential fatty acids have not been reported. Male Sprague-Dawley rats were fed for 12 weeks on pelleted semi-synthetic diets containing less than 1 mg/kg copper (low copper), 6 mg/kg (copper control), or 250 mg/kg copper (copper supplemented). The fatty acid composition of the total phopholipids and triglycerides of plasma, liver, heart and adipose tissue was analyzed by gas liquid chromatography. In low copper rats compared to controls, palmitic and oleic acids were decreased but stearic acid and docosahexaenoic acid were increased in plasma, liver and heart phopholipids. Arachidonic acid was also increased in plasma and liver phospholipids in low copper rats. In liver triglycerides, linoleic and arachidonic acids were increased but palmitic and oleic acid were decreased in low copper rats. Copper supplementation had the opposite effect; palmitic and oleic acids were increased in phospholipids and triglycerides whereas essential fatty acids were generally decreased. Hence, copper not only has a direct effect on the desaturation of stearic acid but also has significant effects on the tissue lipid composition of essential fatty acids.     

Effects of long-chain monounsaturated and n-3 fatty acids on fatty acid oxidation and lipid composition in rats

Long-chain n-3 fatty acids and fat fish are reported, among multiple physiological properties, to enhance peroxisomal beta-oxidation and effect triacylglycerol status. Long-chain n-3 and monounsaturated fatty acids are the main portion of fatty acids in fat fish. The individual effect of long-chain monounsaturated fatty acids on beta-oxidation and fatty acid composition was tested and compared to the effect of n-3 polyunsaturated and saturated fatty acids in a 3-week feeding experiment of rats. To explore the contribution from long-chain monounsaturated fatty acids in these aspects, the effect of long-chain n-3 and monounsaturated fatty acids on mitochondrial and peroxisomal beta-oxidation was compared, as well as fatty acid composition of adipose tissue, liver and serum. Fatty acid oxidase, palmitoyltransferase I and II activities, the amount of serum lipids, and the fatty acid composition of lipid fractions from the organs were analysed. The peroxisomal beta-oxidation was enhanced by the n-3 fatty acids, whereas a small, significant increase with the monounsaturated fatty acids was observed. There was a stimulation of the mitochondrial oxidation with the n-3 fatty acids, but monounsaturated fatty acids gave a small, nonsignificant decrease. With n-3 fatty acids there was a considerable decrease in the levels of serum triacylglycerol, phospholipids, free fatty acids and total cholesterol, while there were only minor effects of monounsaturated fatty acids. As judged from the fatty acid composition data, there was a mobilization on n-3 fatty acids from the adipose tissue to liver and plasma with the n-3 diet. This observation was also seen with the monounsaturated fatty acid-enriched diet. In conclusion, monounsaturated fatty acids seemed to stimulate peroxisomal beta-oxidation and to increase plasma triacylglycerol, whereas the mitochondrial oxidation was slightly decreased.     

Evidence for a role in stimulus--secretion coupling of prostaglandins derived from release of arachidonoyl residues as a result of phosphatidylinositol breakdown.

That stimulation of secretion in exocrine and endocrine glands is associated with increased turnover of phosphatidylinositol and phosphatidic acid has been known for many years. In the present work, mouse pancreases were prelabeled with [14C]arachidonic acid in the presence of the secretogogue carbamoylcholine. They were then incubated in media containing atropine and 1% albumin. The atropine causes the tissue to revert to the resting state, and the albumin binds free [14C]arachidonic acid. The tissues were finally incubated in media containing no stimulant or the stimulant caerulein, which is not blocked by atropine. Stimulation with caerulein, which is not blocked by atropine. Stimulation with caerulein led to a 44% loss of [1-14C]arachidonic acid from phosphatidylinositol. About half of this released arachidonic acid ended up in phosphatidic acid. The remainder of the loss could not be accounted for in any other lipid. No other phospholipids showed statistically significant changes on stimulation. Several lines of evidence indicated that the missing arachidonic acid was converted to prostaglandins, which play a role in stimulus--secretion coupling. Four nonsteroidal anti-inflammatory drugs inhibited secretogogue-induced amylase secretion from pancreases, and their potencies paralleled their potencies in inhibiting cyclooxygenase, which converts arachidonic acid to prostaglandins. Amylase secretion was stimulated by arachidonic acid, and this stimulation was blocked by the nonsteroidal anti-inflammatory drug indomethacin. Other fatty acids failed to elicit amylase secretion. At concentrations of 3--10 nM, prostaglandins I2, E1, E2, D2, and F2 alpha gave statistically significant stimulations of secretion. Other prostaglandins tested gave no significant stimulation.     

The effect of anoxia on lipid metabolism in isolated adult rat cardiac myocytes

In ischemic myocardium abnormal lipid metabolism results in accumulation of compounds that are deleterious to membrane structural integrity and membrane dependent functions. In this study isolated adult rat ventricular myocytes were used to investigate anoxia-induced alterations in cellular lipid composition and metabolism. Myocyte phospholipid content declined 19% on average during 60 min anoxia and intracellular arachidonic acid increased 3-fold, without affecting myocyte ATP content. Anaerobic incubation in the absence of glucose depleted cellular ATP to 2 nmol/mg protein, elicited a 23% decrease in phospholipids, and reduced triacylglycerol content by 51%. Intracellular levels of C16-C22 fatty acids were significantly elevated, especially palmitic and arachidonic acids. Myocytes presented with 0.08 mM [1-14C]-palmitic or arachidonic acid acylated 85% (25-26 nmol/mg) of the fatty acid taken up into triacylglycerols. Anoxia decreased this esterification by 46-60%. Formation of [14C]-CO2 was also depressed 70-90% by anaerobiosis. The results demonstrate that anoxia stimulates degradation of complex lipids, with a concomitant increase in non-esterified fatty acids, especially arachidonic acid.     

Fatty Acid ω- and (ω-1)-Hydroxylation in Experimental Alcoholic Liver Disease: Relationship to Different Dietary Fatty Acids

Arachidonic acid concentrations in liver are decreased in response to ethanol administration. In addition, the oxygenated products of arachidonic acid metabolites could affect the severity of alcoholic liver injury. Selective utilization of arachidonic acid by the cyto-chrome P-450 system could, in part, account for the decrease in arachidonic acid. To evaluate this pathway further, male Wistar rats were fed different dietary fats: medium chain triglycerides, palm oil, and corn oil or fish oil with either ethanol or isocaloric amounts of dextrose. Histopathology, cytochrome P-45ME1 (CYP2E1) and cy-tochrome P-4504A (CYP4A), and ω- and (ω-1)-hydroxylation products of lauric and arachidonic acids were evaluated. Ethanol induction of CYP2El was related to the concentration of polyunsaturated fatty acids in the diet; induction of CYP4A by ethanol was seen in all groups. The highest levels of 11-hydroxy-lauric acid and 19-hydroxyarachidonic acid (ω-1) were seen in rats fed ethanol with palm oil and corn oil. Highly significant correlations were seen between the (ω-1)-hydroxylation products and CYP2E1 activity. No correlation was seen between the ω-hydroxylation products and CYP2E1 activity. In contrast, the levels of ω-hydroxylation products correlated with CYP4A. The overall results showed a significant increase in (ω-1)-hydroxylation products in rats fed diets containing significant amounts of linoleic acid (i.e., palm oil and corn oil).     

Mononuclear phagocytes and eicosanoids: Aspects of their synthesis and biological activities

Summary Mononuclear phagocytes convert arachidonic acid and other unsaturated fatty acids from intracellular sources to a variety of oxygenated metabolites such as prostaglandins and leukotrienes which are secreted into the surrounding medium. Other oxidative products such as hydroxylinoleic acids are reacylated into cellular constituents. The underlying metabolic pathways are activated by numerous stimuli of exogenous or endogenous origin. Depending on the state of activation and cell differentiation, the organ of origin and the nature of the stimulus used, macrophages elaborate a distinct spectrum of oxidative arachidonic acid metabolites. The contribution of these metabolites to the proinflammatory properties of macrophages is twofold: As autocrine signals they modulate the synthesis of diverse macrophage products and they influence cellular functions of other cells such as T-lymphocytes.     

Polyunsaturated fatty acid deficiency reverses effects of alcohol on mitochondrial energy metabolism

BACKGROUND/AIMS: Polyunsaturated fatty acids (PUFA) deficiency is common in patients with alcoholic liver disease. The suitability of reversing such deficiency remains controversial. The aim was to investigate the role played by PUFA deficiency in the occurrence of alcohol-related mitochondrial dysfunction. METHODS: Wistar rats were fed either a control diet with or without alcohol (control and ethanol groups) or a PUFA deficient diet with or without alcohol (PUFA deficient and PUFA deficient+ethanol groups). After 6 weeks, liver mitochondria were isolated for energetic studies and fatty acid analysis. RESULTS: Mitochondria from ethanol fed rats showed a dramatic decrease in oxygen consumption rates and in cytochrome oxidase activity. PUFA deficiency showed an opposite picture. PUFA deficient+ethanol group roughly reach control values, regarding cytochrome oxidase activity and respiratory rates. The relationship between ATP synthesis and respiratory rate was shifted to the left in ethanol group and to the right in PUFA-deficient group. The plots of control and PUFA deficient+ethanol groups were overlapping. Phospholipid arachidonic over linoleic ratio closely correlated to cytochrome oxidase and oxygen uptake. CONCLUSIONS: PUFA deficiency reverses alcohol-related mitochondrial dysfunction via an increase in phospholipid arachidonic over linoleic ratio, which raises cytochrome oxidase activity. Such deficiency may be an adaptive mechanism.