RESUMO
OBJECTIVES: The objective of this study was to examine whether statin therapy is associated with enhanced endothelium-dependent vascular function, improved pulmonary function and reduced systemic inflammation in patients with chronic obstructive pulmonary disease (COPD). DESIGN AND SETTING: This randomized, placebo-controlled, double-blind, parallel trial including patients with COPD was performed at two University hospitals in Norway. SUBJECTS, INTERVENTION AND MEASUREMENTS: Patients with stable COPD (n = 99) were assigned randomly to receive rosuvastatin 10 mg (n = 49) or matching placebo (n = 50) once daily for 12 weeks. The primary outcome measure was change in endothelium-dependent vascular function measured using peripheral arterial tonometry and expressed as the reactive hyperaemia index. Secondary end-points were change in pulmonary function, as assessed by forced expiratory volume in 1 s (FEV1) and FEV1/forced vital capacity (FVC), and change in the circulating levels of the inflammatory markers interleukin-6 (IL6) and high-sensitivity C-reactive protein (hsCRP). RESULTS: In the overall study population, no significant between-group difference in change in endothelium-dependent vascular or pulmonary function was observed. Rosuvastatin therapy was associated with a reduction in hsCRP (-20% vs. 11%, P = 0.017) and an attenuation of the rise in IL6 concentration (8% vs. 30%, P = 0.028) compared with placebo. In a prespecified subgroup analysis of patients with a supra-median circulating hsCRP concentration (>1.7 mg L(-1) ), rosuvastatin was associated with improved endothelium-dependent vascular function (13% vs. 2%, P = 0.026). CONCLUSIONS: In stable COPD patients without the standard indications for statin therapy, rosuvastatin treatment is associated with a significant attenuation of systemic inflammation and improvement in endothelial-dependent vascular function in patients with evidence of systemic inflammation.
Assuntos
Endotélio Vascular/fisiopatologia , Fluorbenzenos/uso terapêutico , Inibidores de Hidroximetilglutaril-CoA Redutases/uso terapêutico , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Doença Pulmonar Obstrutiva Crônica/fisiopatologia , Pirimidinas/uso terapêutico , Sulfonamidas/uso terapêutico , Idoso , Biomarcadores/sangue , Proteína C-Reativa/metabolismo , Método Duplo-Cego , Feminino , Volume Expiratório Forçado , Humanos , Inflamação/fisiopatologia , Interleucina-6/sangue , Pulmão/fisiopatologia , Masculino , Doença Pulmonar Obstrutiva Crônica/sangue , Rosuvastatina Cálcica , Capacidade VitalRESUMO
BACKGROUND: Climate therapy (heliotherapy) of psoriasis is an effective and natural treatment. Ultraviolet radiation (UVB) from the sun improves psoriasis and induces vitamin D(3) synthesis. OBJECTIVE: The aim of the study was to investigate the effect of climate therapy on vitamin D(3) synthesis, blood glucose, lipids and vitamin B12 in psoriasis patients. METHODS: Twenty Caucasian patients (6 women and 14 men; mean age, 47.2 years; range, 24-65) with moderate to severe psoriasis [mean Psoriasis Area and Severity Index (PASI) score 9.8; range, 3.8-18.8] received climate therapy at the Gran Canarias for 3 weeks. Blood samples were drawn before and after 15 days of sun exposure. In addition, the patients' individual skin UV doses based on UV measurements were estimated. RESULTS: Sun exposure for 15 days lead to a 72.8% (+/- 18.0 SD) reduction in the PASI score in psoriasis patients. Although no direct correlation was observed between PASI score improvement and UVB dose, the sun exposure improved the vitamin D, lipid and carbohydrate status of the patients. The serum concentrations of 25-hydroxyvitamin D [25(OH)D] increased from 57.2 +/- 14.9 nmol/L before therapy to 104.5 +/- 15.8 nmol/L (P < 0.0001) after 15 days of sun exposure; the serum levels of 1,25-dihydroxyvitamin D [1,25(OH)(2)D] increased from 146.5 +/- 42.0 to 182.7 +/- 59.1 pmol/L (P = 0.01); the ratio of low-density lipoprotein cholesterol and high-density lipoprotein cholesterol decreased from 2.4 to 1.9 (P < 0.001); and the haemoglobin A(1)c (HbA(1)c) levels decreased from 5.6 +/- 1.7% to 5.1 +/- 0.3% (P < 0.0001). CONCLUSION: Climate therapy with sun exposure had a positive effect on psoriasis, vitamin D production, lipid and carbohydrate status.
Assuntos
Glicemia/análise , Helioterapia , Lipídeos/sangue , Psoríase/terapia , Vitamina D/biossíntese , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Psoríase/sangue , Raios Ultravioleta , Vitamina D/análogos & derivados , Vitamina D/sangueRESUMO
INTRODUCTION: The CellaVision™DM96 is a digital pattern recognition system that classifies white blood cells. The aim of this study was to evaluate whether the CellaVision preclassification feature, without a subsequent re-classification, was a sufficient approach to follow up flags reported by Sysmex XE-5000. METHODS: Pairs of blood smears from 400 samples reported with suspect flags were examined using conventional microscopy and the CellaVision features. The effect of pre- vs. re-classification, and intersmear and between-technologist variation, on blast counts was assessed using generalized linear mixed models (GLMM). RESULTS: The GLMM analysis showed a significant difference between the blast counts of preclassification vs. re-classification (P = 0.009). The analysis showed no significant difference between duplicate smears (P = 0.621) or between technologists (P = 0.542). Preclassification showed blasts in 105 samples (26%), where the re-classification feature did not detect any blasts. Not a single sample that was re-classified as positive for blasts was preclassified as negative. Compared to manual microscopy, the sensitivity and specificity of the preclassification feature were 0.83 and 0.66, respectively. CONCLUSION: The preclassification feature alone is sufficient to verify the absence of blasts in flagged samples. When the preclassification feature detects blasts, the finding has to be verified or reclassified by an experienced technologist. However, the use of CellaVision™DM96 in the follow-up of blast reports has to be questioned due to the finding of false-negative samples in the preclassification feature, but also after re-classification, compared to manual slide review.
Assuntos
Contagem de Leucócitos/métodos , Contagem de Leucócitos/normas , Leucócitos/patologia , Humanos , Contagem de Leucócitos/instrumentação , Microscopia , Curva ROC , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
The intracellular localization of the oxidation of [2-14C]adrenic acid (22:4(n-6)) and [1-14C]docosahexaenoic acid (22:6(n-3)) was studied in isolated liver cells. The oxidation of 22:4(n-6) was 2-3-times more rapid than the oxidation of 22:6(n-3), [1-14C]arachidonic acid (20:4(n-6)) or [1-14C]oleic acid (18:1). (+)-Decanoylcarnitine and lactate, both known to inhibit mitochondrial beta-oxidation, reduced the oxidation of 18:1 distinctly more efficiently than with 22:4(n-6) and 22:6(n-3). In liver cells from rats fed a diet containing partially hydrogenated fish oil, the oxidation of 22:6(n-6) and 22:6(n-3) was increased by 30-40% compared with cells from rats fed a standard pellet diet. With 18:1 as substrate, the amount of fatty acid oxidized was very similar in cells from animals fed standard pellets or partially hydrogenated fish oil. Shortened fatty acids were not produced from [5,6,8,9,11,12,14,15-3H]arachidonic acid. In hepatocytes from rats starved and refed 20% fructose, a large fraction of 14C from 22:4 was recovered in 14C-labelled C14-C18 fatty acids. Oxidation of 22:4 thus caused a high specific activity of the extramitochondrial pool of acetyl-CoA. The results suggest that 22:4(n-6) and to some extent 22:6(n-3) are oxidized by peroxisomal beta-oxidation and by this are retroconverted to arachidonic acid and eicosapentaenoic acid.
Assuntos
Ácidos Erúcicos/metabolismo , Ácidos Graxos Insaturados/metabolismo , Fígado/citologia , Microcorpos/metabolismo , Animais , Carnitina/análogos & derivados , Carnitina/farmacologia , Gorduras na Dieta/administração & dosagem , Ácidos Docosa-Hexaenoicos , Jejum , Lactatos/farmacologia , Ácido Láctico , Fígado/metabolismo , Masculino , Ácido Oleico , Ácidos Oleicos/farmacologia , Ratos , Ratos EndogâmicosRESUMO
When [14C]linoleic acid (18:2(n-6)) or [14C]dihomogammalinolenic acid (20:3(n-6)) was incubated with isolated liver cells from rats fed an essential fatty acid deficient diet, delta 6- and delta 5-desaturation, chain elongation and synthesis of 14C-labelled C14-C18 fatty acids (from [14C]acetate) were enhanced in female cells compared with male ones. No sex difference in total secretion of very low density lipoproteins (VLDL) was observed. However, VLDL secreted from female cells contained significantly more C16-C18 fatty acids than male cells. It is suggested that the observed sex differences, at least in part, may be related to the different content of fatty acid binding proteins in female cells compared with males.
Assuntos
Ácidos Graxos Dessaturases/metabolismo , Ácidos Graxos Essenciais/metabolismo , Fígado/metabolismo , Caracteres Sexuais , Ácido 8,11,14-Eicosatrienoico , Acetatos/metabolismo , Ácido Acético , Animais , Ácidos Graxos Essenciais/deficiência , Feminino , Ácido Linoleico , Ácidos Linoleicos/metabolismo , Ácidos Linolênicos/metabolismo , Lipoproteínas VLDL/metabolismo , Masculino , Ratos , Ratos EndogâmicosRESUMO
The triacylglycerol synthesis from exogenous linoleic acid (18:2(n-6], linolenic acid (18:3(n-3], dihomogammalinolenic acid (20:3(n-6], eicosapentaenoic acid (20:5(n-3] and oleic acid (18:1(n-9] was observed to be significantly increased in isolated liver cells from female rats compared with males. The rate of fatty acid oxidation and phospholipid biosynthesis was concomitantly more important in male cells. With the C22-polyenoic fatty acids, adrenic acid (22:4(n-6] and docosahexaenoic acid (22:6(n-3), only a minor sex-related difference in fatty acid metabolism was found.
Assuntos
Ácidos Graxos Essenciais/metabolismo , Fígado/metabolismo , Animais , Esterificação , Feminino , Técnicas In Vitro , Masculino , Oxirredução , Ratos , Ratos Endogâmicos , Fatores SexuaisRESUMO
alpha-Bromopalmitate was shown to have a far more pronounced effect on metabolism of labelled linoleic acid (18:2, n-6) and arachidonic acid (20:4, n-6) in isolated liver cells from female rats than in those from males. alpha-Bromopalmitate decreased triacylglycerol synthesis with a concomitant accumulation of fatty acid in diacylglycerol, indicating that the acylation of diacylglycerol is affected by alpha-bromopalmitate.
Assuntos
Ácidos Graxos Essenciais/metabolismo , Fígado/metabolismo , Palmitatos/farmacologia , Ácidos Palmíticos/farmacologia , Animais , Ácido Araquidônico , Ácidos Araquidônicos/metabolismo , Esterificação , Feminino , Técnicas In Vitro , Ácido Linoleico , Ácidos Linoleicos/metabolismo , Fígado/efeitos dos fármacos , Masculino , Oxirredução/efeitos dos fármacos , Ratos , Caracteres SexuaisRESUMO
The uptake and integrated intracellular metabolism of (n - 6) and (n - 3) polyunsaturated fatty acids was studied in isolated rat cardiac myocytes and in the perfused heart. Labeled linolenic acid (18:3(n - 3)) uptake and its subsequent metabolism into carbon dioxide as well as acylation into lipids was nonsaturable over a substrate range of 0.02 to 0.4 mM. [1-14C]Linoleic acid (18:2(n - 6)), dihomo-gamma-linolenic acid (20:3(n - 6)) and arachidonic acid (20:4(n - 6)) were transported into myocytes at rates similar to those for linolenic acid. Conversely both [1-14C]-gamma-linolenic acid (18:3(n - 6)) and eicosapentaenoic acid (20:5(n - 3)) were taken up at a slower rate. Oxidation of 18:3(n - 6) was 4-5-fold greater when compared with C18-C20 polyunsaturated fatty acids. When myocytes were incubated with labeled 18:2(n - 6), 18:3(n - 6), 18:3(n - 3), 20:4(n - 6) or 20:5(n - 3), it was not possible to detect any desaturation or chain-elongation products. Identical results were obtained when hearts were perfused with 1-14C-labeled linoleic acid.
Assuntos
Ácidos Graxos Insaturados/metabolismo , Miocárdio/metabolismo , Animais , Ácidos Graxos Insaturados/farmacocinética , Ácidos Linolênicos/farmacocinética , Masculino , Ratos , Ratos Endogâmicos , Relação Estrutura-Atividade , Fatores de Tempo , Ácido alfa-LinolênicoRESUMO
The desaturation and chain elongation of [1-14C]linolenic acid was studied in isolated liver cells from rats fed a diet deficient in essential fatty acids. 14C-labelled 18:4, 20:3, 20:4, 20:5, 22:5 and 22:6, all n - 3 fatty acids, were formed. In the presence of lactate relatively large amounts of 20:5, 22:5 and 22:6 were formed. 20:5 was mainly present in phospholipids, 22:5 and 22:6 were present in both phospholipids and triacylglycerols. (+)-Decanoylcarnitine and (-)-hydroxycitrate decreased the formation of 20:5, 22:5 and 22:6 and increased the recovery of 18:4. The unchanged 18:3 substrate was also initially rapidly incorporated both in the phospholipids and in the triacylglycerol fraction. During long incubation periods, continued after nearly all the [14C]linolenic acid substrate had been metabolized either by esterification or by oxidation, the phospholipid content of labelled 18:3 and 18:4 decreased while the content of 20:5, 22:5 and 22:6 increased markedly, suggesting a remodeling of the phospholipid n - 3 fatty acid content by a series of deacylations-reacylations. The n - 3 fatty acid pattern in the triacylglycerol fraction changed little. 22:5 and 22:6 appeared in the VLDL fraction secreted by the isolated liver cells.
Assuntos
Ácidos Graxos/metabolismo , Ácidos Linolênicos/metabolismo , Fígado/metabolismo , Fosfolipídeos/metabolismo , Animais , Esterificação , Ácidos Graxos Dessaturases/metabolismo , Técnicas In Vitro , Fígado/enzimologia , Masculino , Oxirredução , Ratos , Ratos EndogâmicosRESUMO
The desaturation, chain elongation and esterification of [1-14C]eicosapentaenoic acid, [1-14C]arachidonic acid, [1-14C]eicosatrienoic acid, [1-14C]linolenic acid and [1-14C]linoleic acid were studied in isolated liver cells. Rats fed diets with either 15% hydrogenated coconut oil or 15% partially hydrogenated marine oil, both deficient in essential fatty acids, 15% soybean oil or standard pellet diet with 6% fat, were used. The delta 4-desaturation of 22:5(n - 3) and 22:4(n - 6) as well as the delta 6-desaturase activity was distinctly higher in cells from animals fed coconut or marine oil than with soybean oil or standard pellet. The rate of delta 5-desaturation of 20:3(n - 6) and 20:4(n - 3) was nearly the same in cells from rats fed coconut, marine and soybean oils and higher than with standard pellet. The chain elongation of 20:5(n - 3) to 22:5(n - 3) was distinctly more pronounced than the elongation of 20:4(n - 6) with all four diets. 20:5(n - 3) was mainly esterified in the phospholipids with marine and coconut oils, and mainly in triacylglycerol with standard pellet and soybean oils. The proportion of [1-14C]20:4(n - 6) in the phospholipids to that in triacylglycerol decreased in the order marine oil greater than coconut oil greater than standard pellet greater than soybean oil. The different endogenous arachidonic acid content in the phospholipids induced by the different diets increased in the same order. 20:5(n - 3) was rapidly esterified in triacylglycerol and phospholipids, then liberated especially from the triacylglycerol fraction, chain elongated to 22:5(n - 3) and reesterified.
Assuntos
Ácidos Araquidônicos/metabolismo , Gorduras na Dieta/farmacologia , Ácidos Graxos Insaturados/metabolismo , Ácidos Linoleicos/biossíntese , Ácidos Linolênicos/biossíntese , Fígado/metabolismo , Animais , Ácido Araquidônico , Radioisótopos de Carbono , Ácido Eicosapentaenoico , Ácidos Graxos/análise , Técnicas In Vitro , Ácido Linoleico , Fígado/efeitos dos fármacos , Masculino , Fosfolipídeos/biossíntese , Ratos , Ratos Endogâmicos , Triglicerídeos/biossíntese , Ácido alfa-LinolênicoRESUMO
In the Zellweger syndrome where peroxisomes are absent, extremely long fatty acids (24:0 and 26:0) accumulate in tissues suggesting that these fatty acids are normally beta-oxidized in the peroxisomes. Previous studies with rat hepatocytes suggest that peroxisomes are also important in oxidation of C22 unsaturated fatty acids. This study shows that cultured fibroblasts from normal human controls shorten [14-14C]erucic acid (22:1(n-9)) to oleic acid (18:1(n-9)) efficiently while Zellweger fibroblasts are deficient in chain-shortening. [2-14C]Adrenic acid (22:4(n-6)) is oxidized in control fibroblasts probably by chain-shortening to arachidonic acid (20:4(n-6)). Only a little adrenic acid is oxidized in Zellweger fibroblasts. Linolenic acid (18:3(n-3)) is desaturated and chain-elongated in both control and Zellweger fibroblasts. The results support the view that peroxisomes play a normal physiological role in the shortening of C22 unsaturated fatty acids and that this function is deficient in Zellweger fibroblasts.
Assuntos
Ácidos Erúcicos/metabolismo , Ácidos Graxos Monoinsaturados/metabolismo , Erros Inatos do Metabolismo Lipídico/metabolismo , Microcorpos/metabolismo , Células Cultivadas , Ácidos Graxos/metabolismo , Ácidos Graxos Insaturados , Humanos , Técnicas In Vitro , Oxirredução , SíndromeRESUMO
The partitioning between peroxisomal and mitochondrial beta-oxidation of [1-14C]eicosapentaenoic acid (20:5(n-3] and [1-14C]arachidonic acid (20:4(n-6)) was studied. In hepatocytes from fasted rats approximately 70% of the fatty acid substrate was oxidized with oleic, linoleic, eicosapentaenoic and docosahexaenoic (22:6(n-3)) acid, even more with adrenic (22:4(n-6)) and less with arachidonic acid. When the mitochondrial oxidation was suppressed by fructose refeeding and by (+)-decanoylcarnitine, the fatty acid oxidation in per cent of that in cells from fasted rats was with 18:1(n-9) 7%, 18:2(n-6) 8%, 20:4(n-6) 12%, 20:5(n-3) 20%, 22:4(n-6) 57% and for 22:6(n-3) 29%. The fraction of 14C recovered in palmitate and other newly synthesized fatty acids after fructose refeeding decreased in the order 22:4(n-6) greater than 22:6(n-3) greater than 20:5(n-3) greater than 20:4(n-6) and was very small with 18:1(n-9) and 18:2(n-6). In cells from both fed and fructose-refed animals 20:5(n-3) was efficiently elongated to 22:5(n-3) and 22:6(n-3). 20:5(n-3) and 20:4(n-6) were not elongated after fasting. The phospholipid incorporation with [1-14C]20:5(n-3) decreased during prolonged incubations while it remained stable with [1-14C]arachidonic acid. The results suggest that peroxisomes contribute more to the oxidation of 20:5(n-3) than with 20:4(n-6) although both substrates are probably oxidized mainly in the mitochondria.
Assuntos
Ácidos Araquidônicos/metabolismo , Ácido Eicosapentaenoico/metabolismo , Fígado/metabolismo , Microcorpos/metabolismo , Mitocôndrias Hepáticas/metabolismo , Mitocôndrias/metabolismo , Animais , Ácido Araquidônico , Radioisótopos de Carbono , Carnitina/análogos & derivados , Carnitina/farmacologia , Técnicas In Vitro , Cinética , Fígado/citologia , Fígado/efeitos dos fármacos , Masculino , Ratos , Ratos EndogâmicosRESUMO
In order to study the effect of n-3 fatty acids on the physical state of the erythrocyte membrane, measured as osmotic fragility, rats were fed a diet supplemented in n-3 fatty acids (1.5 ml/day, 35% 20:5, 30% 22:6) for 21 days. With salt concentrations ranging from 0.37% to 0.44%, osmotic resistance was increased by 25% to 45% in cells from n-3-fed animals compared to controls. No change was observed in either phospholipid or cholesterol content in the membrane. A small, but still significant difference (P less than 0.05) in phospholipid sub-class distribution was observed in that the phosphatidylethanolamine fraction was decreased and the phosphatidylserine fraction increased after n-3 supplementation. The major change was, however, that the level of eicosapentaenoic acid (20:5(n-3] in phospholipids was increased from 1.5% of total fatty acids to 4.5%. This increase was mainly at the expense of linoleic acid (18:2(n-6]. No change was observed in the level of docosahexaenoic acid (22:6(n-3]. It is thus concluded that both the fatty acid composition and the nature of the phospholipid polar head group may influence the osmotic fragility of erythrocytes.
Assuntos
Membrana Eritrocítica/efeitos dos fármacos , Ácidos Graxos Insaturados/farmacologia , Fragilidade Osmótica/efeitos dos fármacos , Animais , Gorduras na Dieta/administração & dosagem , Membrana Eritrocítica/química , Ácidos Graxos Insaturados/administração & dosagem , Masculino , Pressão Osmótica , Fosfolipídeos/sangue , Ratos , Ratos EndogâmicosRESUMO
Isolated liver cells from rats fed a diet deficient in essential fatty acids were used to study the oxidation, esterification and, especially, the desaturation and chain elongation of [1-14C]linoleic acid. 14C-labelled arachidonic acid (20:4) and smaller amounts of eicosatrienoic acid (20:3) were recovered mainly in the phospholipids, while gamma-linolenic acid (18:3) was found in both the phospholipids and the triacylglycerol fraction. Lactate strongly increased the formation of arachidonic acid, which was found mainly in the phosphatidylcholine and the phosphatidylinositol fractions. Lactate reduced the amounts of gamma-linolenic acid. Glucagon and (+)-decanoylcarnitine reduced the formation of arachidonic acid, and (+)-decanoylcarnitine increased the incorporation of gamma-linolenic acid especially, in the triacylglycerol fraction. Increasing concentrations of the [1-14C]linoleic acid substrate increased the formation of arachidonic acid and of the other chain-elongated or desaturated fatty acids. Lactate also stimulated the formation of arachidonic acid in liver cells from animals fed adequate amounts of essential fatty acids. It is suggested that dietary and hormonal factors which can change the intracellular levels of malonyl-CoA may influence both the ratio of arachidonic acid/gamma-linolenic acid formed and the total amounts of desaturated and chain-elongated fatty acids formed from linoleic acid.
Assuntos
Ácidos Araquidônicos/biossíntese , Fígado/metabolismo , Animais , Ácido Araquidônico , Ácidos Graxos Insaturados/metabolismo , Glucagon/farmacologia , Lactatos/farmacologia , Ácido Láctico , Ácido Linoleico , Ácidos Linoleicos/metabolismo , Masculino , Malonil Coenzima A/metabolismo , Ratos , Ratos Endogâmicos , Fatores de TempoRESUMO
The metabolism of docosahexaenoic acid (22:6(n-3)) and adrenic acid (22:4(n-6)) was studied in cultured fibroblasts from patients with the Zellweger syndrome, X-linked adrenoleukodystrophy (X-ALD) and normal controls. It was shown that [4,5- 3H]22:6(n-3) is retroconverted to labelled eicosapentaenoic acid (20:5(n-3)) in normal and X-ALD fibroblasts, while this conversion is deficient in Zellweger fibroblasts. [U- 14C]Eicosapentaenoic acid (20:5(n-3)) is elongated to docosapentaenoic acid (22:5(n-3)) in all three cell lines. With [U- 14C]20:5(n-3) as the substrate, shorter fatty acids were not detected. With [4,5- 3H]22:6(n-3) as the substrate, labelled fatty acids were esterified in the phospholipid- and triacylglycerol-fraction to approximately the same extent in all three cell lines. [2- 14C]Adrenic acid (22:4(n-6)) was desaturated to 22:5(n-6) and elongated to 24:4(n-6) in all three cell lines and to the largest extent in the Zellweger fibroblasts. This agrees with the view that the delta 4-desaturase is not a peroxisomal enzyme. The observation that the retroconversion of 22:6(n-3) to 20:5(n-3) is deficient in Zellweger fibroblasts strongly suggest that the beta-oxidation step in the retroconversion is a peroxisomal function. Peroxisomal very-long-chain (lignoceroyl) CoA ligase is probably not required for the activation of 22:6(n-3), since the retroconversion to 20:5(n-3) is normal in X-ALD fibroblasts.
Assuntos
Ácidos Docosa-Hexaenoicos/metabolismo , Ácido Eicosapentaenoico/metabolismo , Ácidos Graxos Dessaturases/metabolismo , Síndrome de Zellweger/metabolismo , Adrenoleucodistrofia/metabolismo , Células Cultivadas , Fibroblastos , Humanos , Linoleoil-CoA Desaturase , Síndrome de Zellweger/enzimologiaRESUMO
The omega- and beta-oxidation of medium- and long-chain fatty acids (C10-C18) were studied in hepatocytes from fasted, fed and clofibrate-fed rats. The omega-oxidation systems were most active with lauric acid (12:0) and decanoic acid (10:0) as substrates and there was decreasing activity with chain lengths from 14 to 18 carbon atoms. In fed rats no omega-oxidation of fatty acids was detected unless the mitochondrial beta-oxidation was inhibited. In fasted rats the omega-oxidation was less than 2% and preincubation with (+)-decanoylcarnitine increased the omega-oxidation to 15% of the total fatty acid oxidation. Clofibrate feeding did not increase the omega-oxidation in isolated hepatocytes. Inhibition of the alcohol dehydrogenase with 4-methylpyrazole inhibited both the oxidation of omega-hydroxylated fatty acid and the initial hydroxylation of lauric acid to dicarboxylic acid, suggesting the importance of the alcohol dehydrogenase in the omega-oxidation of fatty acids. 95% of the dicarboxylic acids and 80% of the hydroxy-fatty acids were excreted from the cells in the incubations with decanoic acid (10:0). No chain-shortened dicarboxylic acids were detected with [1-14C]decanoic- or [1-14C]lauric acid as substrate, while small amounts C10 and C12 dicarboxylic acids were observed in incubations with [1-14C]myristic acid (14:0).
Assuntos
Ácidos Graxos/metabolismo , Mitocôndrias Hepáticas/metabolismo , Álcool Desidrogenase/antagonistas & inibidores , Animais , Carnitina/análogos & derivados , Carnitina/farmacologia , Hidrólise , Técnicas In Vitro , Masculino , Mitocôndrias Hepáticas/efeitos dos fármacos , Mitocôndrias Hepáticas/enzimologia , Oxirredução , Ratos , Ratos EndogâmicosRESUMO
The beta-oxidation and esterification of medium-chain fatty acids were studied in hepatocytes from fasted, fed and fructose-refed rats. The beta-oxidation of lauric acid (12:0) was less inhibited by fructose refeeding and by (+)-decanoyl-carnitine than the oxidation of oleic acid was, suggesting a peroxisomal beta-oxidation of lauric acid. Little lauric acid was esterified in triacylglycerol fraction, except at high substrate concentrations or in the fructose-refed state. With [1-14C]myristic acid (14:0), [1-14C]lauric acid (12:0), [1-14C]octanoic acid (8:0) and [2-14C]adrenic acid (22:4(n - 6] as substrate for hepatocytes from carbohydrate-refed rats, a large fraction of the 14C-labelled esterified fatty acids consisted of newly synthesized palmitic acid (16:0), stearic acid (18:0) and oleic acid (18:1) while intact [1-14C]oleic acid substrate was esterified directly. With [9,10-3H]myristic acid as the substrate, small amounts of shortened 3H-labelled beta-oxidation intermediates were found. With [U-14C]palmitic acid, no shortened fatty acids were detected. It was concluded that when the mitochondrial fatty acid oxidation is down-regulated such as in the carbohydrate-refed state, medium-chain fatty acids can partly be retailored to long-chain fatty acids by peroxisomal beta-oxidation followed by synthesis of C16 and C16 fatty acids which can then stored as triacylglycerol.
Assuntos
Ácidos Graxos/metabolismo , Fígado/metabolismo , Animais , Caprilatos/metabolismo , Carnitina/análogos & derivados , Carnitina/farmacologia , Esterificação , Jejum , Alimentos , Frutose/farmacologia , Ácidos Láuricos/metabolismo , Fígado/efeitos dos fármacos , Masculino , Ácido Mirístico , Ácidos Mirísticos/metabolismo , Ácido Oleico , Ácidos Oleicos/metabolismo , Oxirredução , Ácido Palmítico , Ácidos Palmíticos/metabolismo , Ratos , Ratos Endogâmicos , Ácidos Esteáricos/metabolismo , Triglicerídeos/metabolismoRESUMO
The effects of dietary supplementation of eicosapentaenoic acid (20:5(n-3), EPA) and docosahexaenoic acid (22:6(n-3), DHA) on the metabolism of polyunsaturated fatty acids were studied in isolated rat liver cells. Both pure EPA and pure DHA and a mixture of the two n-3 fatty acids in different doses were used. The supplementation of moderate amounts of n-3 fatty acids suppressed the activity of delta 6-desaturase (50%) and to a smaller extent of the delta 5-desaturase (60-70%) compared to controls. When higher doses of dietary purified EPA and DHA were used, this inhibitory effect on the delta 6- and delta 5-desaturase activities disappeared. The delta 4-desaturase activity seemed to be unaffected by the feeding conditions used. The supplementation of the n-3 fatty acids in the diet at all dose levels used increased the beta-oxidation of all the polyunsaturated fatty acids, especially of linoleic acid, linolenic acid and eicosapentaenoic acid. The results suggest an increase both in peroxisomal and mitochondrial beta-oxidation. The peroxisomal beta-oxidation of n-3 fatty acids seemed to be particularly increased.
Assuntos
Gorduras na Dieta/farmacologia , Ácidos Docosa-Hexaenoicos/farmacologia , Ácido Eicosapentaenoico/farmacologia , Ácidos Graxos/metabolismo , Fígado/metabolismo , Animais , Células Cultivadas , Ácido Eicosapentaenoico/isolamento & purificação , Esterificação , Fígado/citologia , Masculino , Oxirredução , Fosfolipídeos/metabolismo , Ratos , Ratos Endogâmicos , Triglicerídeos/metabolismoRESUMO
The effects of clofibrate feeding on the metabolism of polyunsaturated fatty acids were studied in isolated rat hepatocytes. Administration of clofibrate stimulated the oxidation and particularly the peroxisomal beta-oxidation of all the fatty acids used. The increase in oxidation products was markedly higher when n-3 fatty acids were used as substrate, indicating that peroxisomes contribute more to the oxidation of n-3 than n-6 fatty acids. The whole increase in oxidation could be accounted for by a corresponding decrease in acylation in triacylglycerol while the esterification in phospholipids remained unchanged. A marked stimulation of the amounts of newly synthesized C16 and C18 fatty acids recovered, was observed when 18:2(n-6), 20:3(n-6), 18:3 (n-3) and 20:5(n-3), but not when 20:4(n-6) and 22:4(n-6) were used as substrate. This agrees with the view that extra-mitochondrial acetyl-CoA produced from peroxisomal beta-oxidation is more easily used for fatty acid new synthesis than acetyl-CoA from mitochondrial beta-oxidation. The delta 6 and delta 5 desaturase activities were distinctly higher in cells from clofibrate fed rats indicating a stimulating effect.
Assuntos
Clofibrato/farmacologia , Ácidos Graxos Dessaturases/metabolismo , Ácidos Graxos Insaturados/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Fígado/metabolismo , Animais , Dessaturase de Ácido Graxo Delta-5 , Gorduras na Dieta/farmacologia , Linoleoil-CoA Desaturase , Fígado/efeitos dos fármacos , Fígado/enzimologia , Masculino , Ratos , Ratos EndogâmicosRESUMO
Retroconversion of docosahexaenoic acid (DHA, 22:6(n-3)) to eicosapentaenoic acid (EPA, 20:5(n-3)) was studied in isolated rat liver cells. 20% of the substrate was retroconverted to EPA in control cells by one cycle of beta-oxidation probably with delta 4 enoyl CoA reductase and delta 3, delta 2 enoyl CoA isomerase as auxiliary enzymes. This conversion was not stimulated by (-)-carnitine and was not inhibited by the addition of (+)-decanoylcarnitine. In hepatocytes from fasted rats little EPA was formed from DHA. These results strongly suggest that the retroconversion of DHA to EPA is a peroxisomal function. Retroconverted EPA, produced from DHA was rapidly incorporated in triacylglycerol, the phosphatidylcholine and phosphatidyletanolamine fractions. During longer incubation time EPA was partly removed from the phospholipid fractions, chain-elongated to 22:5(n-3) and incorporated in the triacylglycerol fraction.