RESUMO
Micronutrients (folates and vitamin B12) and long chain polyunsaturated fatty acids (LC-PUFAs) are linked through the one carbon cycle. We studied the effects of pre and postnatal high FA/low B12 diets (HFLB12) on hepatic fatty acid metabolism. Pregnant C57BL/6 mice were divided in two groups: control (2 mg folic acid: FA/25 µg vitamin B12/Kg food) and HFLB12 diets (8 mg FA/5 µg vitamin B12/Kg food). Offspring continued on the same diets until 60 days old. We determined hepatic fatty acid profile in dams and offspring and the expression of PPARα, Cpt-1, Acox-1 and Fas and the enzymatic activity of desaturases, all involved in lipid metabolism. In liver of dams, the HFHB12 diet decreased total fatty acids and desaturase activities; in offspring, effects were opposite, being more noticeable in females. Prenatal and postnatal unbalanced folic acid/B12 diets play a crucial role in regulating genes and enzymes involved in lipid metabolism in liver of dams and their offspring in adulthood.
Assuntos
Ácidos Graxos/metabolismo , Ácido Fólico/administração & dosagem , Metabolismo dos Lipídeos/efeitos dos fármacos , Fígado/química , Vitamina B 12/administração & dosagem , Acil-CoA Oxidase/metabolismo , Animais , Animais Recém-Nascidos , Ácidos Graxos Dessaturases/metabolismo , Feminino , Ácido Fólico/farmacocinética , Regulação da Expressão Gênica/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , PPAR alfa/metabolismo , Cuidado Pós-Natal , Gravidez , Vitamina B 12/farmacocinética , Receptor fasRESUMO
Remains unknown if dietary lipids and anabolic steroids (AS) can interact to modify energy metabolism, hepatic structure and function. We investigated the impact of AS on gene expression, lipid profile, redox status and the development of nonalcoholic fatty liver disease (NAFLD) in mice treated with a diet rich in trans fatty acids. Seventy-two C57BL/6 mice were equally randomized into six groups and treated with a standard diet (SD) or high-fat diet (HFD) alone or combined with testosterone cypionate (10 or 20â¯mg/kg) for 12 weeks. When combined with a HFD, AS reduced plasma HDL cholesterol levels. It also upregulated SREBP-1, PPARα, SCD-1 and ACOX1 gene expression; plasma and hepatic triglyceride levels; oxidative stress; circulating hepatic transaminase levels and NAFLD severity. Our finding indicated that the activity of antioxidant enzymes such as catalase, glutathione-s-transferase and superoxide dismutase was attenuated by HFD, an effect whose implications for AS-induced hepatotoxicity requires further investigation. Increased lipid, protein and DNA oxidative damage as well as worsening NAFLD in response to the interaction of HFD and AS were also potentially associated with the ability of AS to amplify the activation of regulatory lipid metabolism genes that are also involved in the control of cellular redox balance.
Assuntos
Interações Alimento-Droga , Fígado/efeitos dos fármacos , Hepatopatia Gordurosa não Alcoólica/induzido quimicamente , Hepatopatia Gordurosa não Alcoólica/fisiopatologia , Congêneres da Testosterona/toxicidade , Ácidos Graxos trans/toxicidade , Triglicerídeos/metabolismo , Acil-CoA Oxidase/genética , Alanina Transaminase/sangue , Animais , Antioxidantes/metabolismo , Aspartato Aminotransferases/sangue , Composição Corporal , Catalase/sangue , Dieta Hiperlipídica , Regulação da Expressão Gênica , Glutationa Transferase/sangue , Fígado/metabolismo , Camundongos Endogâmicos C57BL , Hepatopatia Gordurosa não Alcoólica/genética , Hepatopatia Gordurosa não Alcoólica/metabolismo , Estresse Oxidativo/efeitos dos fármacos , PPAR alfa/genética , Estearoil-CoA Dessaturase/genética , Proteína de Ligação a Elemento Regulador de Esterol 1/genética , Superóxido Dismutase/sangue , Triglicerídeos/sangue , Regulação para CimaRESUMO
SCOPE: Dietary n-3 long-chain PUFAs (n-3 LCPUFAs) supplementation was studied in an HFD-induced (HFD is high-fat diet) steatosis and inflammation in relation to peroxisome proliferator-activated receptor alpha (PPAR-α) and nuclear factor κB (NF-κB) signaling. METHODS AND RESULTS: Male C57BL/6J mice received (i) control diet (10% fat, 20% protein, 70% carbohydrate), (ii) control diet plus n-3 LCPUFAs (daily doses of 108 mg/kg body weight of eicosapentaenoic acid plus 92 mg/kg body weight of docosahexaenoic acid), (iii) HFD (60% fat, 20% protein, 20% carbohydrate), or (iv) HFD plus n-3 LCPUFAs for 12 wk. PPAR-α, tumor necrosis factor alpha (TNF-α), and IL-1ß mRNA expression, acyl-CoA oxidase 1 (ACOX1), and carnitine-acyl-CoA transferase 1 (CAT-I) protein contents, and NF-κB DNA binding activity were measured. HFD significantly decreased liver PPAR-α, ACOX1, and CAT-I levels with NF-κB activation, higher TNF-α and IL-1ß expression, and steatosis development. These changes were either reduced or normalized to control values in animals subjected to HFD plus n-3 LCPUFAs, with establishment of an inverse association between NF-κB activation and PPAR-α mRNA expression (r = -0.66, p < 0.0001). CONCLUSION: Data presented indicate that n-3 LCPUFAs supplementation prevents liver steatosis and inflammation induced by HFD, with underlying mechanisms involving enhanced PPAR-α signaling and diminished NF-κB activation.
Assuntos
Dieta Hiperlipídica/efeitos adversos , Suplementos Nutricionais , Ácidos Graxos Ômega-3/administração & dosagem , Fígado Gorduroso/prevenção & controle , NF-kappa B/metabolismo , PPAR alfa/metabolismo , Acil-CoA Oxidase/genética , Acil-CoA Oxidase/metabolismo , Animais , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Fígado Gorduroso/etiologia , Inflamação/etiologia , Inflamação/prevenção & controle , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Fígado/enzimologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , NF-kappa B/genética , Tamanho do Órgão , PPAR alfa/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Regulação para CimaRESUMO
Estrogen deficiency accelerates the development of several disorders including visceral obesity and hepatic steatosis. The predisposing factors can be exacerbated by drugs that affect hepatic lipid metabolism. The aim of the present work was to determine if raloxifene, a selective estrogen receptor modulator (SERM) used extensively by postmenopausal women, affects hepatic fatty acid oxidation pathways. Fatty acids oxidation was measured in the livers, mitochondria and peroxisomes of ovariectomized (OVX) rats. Mitochondrial and peroxisomal ß-oxidation was inhibited by raloxifene at a concentration range of 2.5-25 µM. In perfused livers, raloxifene reduced the ketogenesis from endogenous and exogenous fatty acids and increased the ß-hydroxybutyrate/acetoacetate ratio. An increase in ¹4CO2 production without a parallel increase in the oxygen consumption indicated that raloxifene caused a diversion of NADH from the mitochondrial respiratory chain to another oxidative reaction. It was found that raloxifene has a strong ability to react with H2O2 in the presence of peroxidase. It is likely that the generation of phenoxyl radical derivatives of raloxifene in intact livers led to the co-oxidation of NADH and a shift of the cellular redox state to an oxidised condition. This change can perturb other important liver metabolic processes dependent on cellular NADH/NAD⺠ratio.
Assuntos
Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Ácidos Graxos/metabolismo , Fígado Gorduroso/induzido quimicamente , Fígado/efeitos dos fármacos , Oxidantes/efeitos adversos , Cloridrato de Raloxifeno/efeitos adversos , Moduladores Seletivos de Receptor Estrogênico/efeitos adversos , Acil Coenzima A/metabolismo , Acil-CoA Oxidase/metabolismo , Animais , Doença Hepática Induzida por Substâncias e Drogas/enzimologia , Doença Hepática Induzida por Substâncias e Drogas/fisiopatologia , Progressão da Doença , Terapia de Reposição de Estrogênios/efeitos adversos , Fígado Gorduroso/metabolismo , Fígado Gorduroso/fisiopatologia , Feminino , Peróxido de Hidrogênio/química , Fígado/enzimologia , Fígado/metabolismo , Mitocôndrias Hepáticas/efeitos dos fármacos , Mitocôndrias Hepáticas/enzimologia , Mitocôndrias Hepáticas/metabolismo , Ovariectomia/efeitos adversos , Oxidantes/química , Oxirredução , Peroxidase/metabolismo , Peroxissomos/efeitos dos fármacos , Peroxissomos/enzimologia , Peroxissomos/metabolismo , Cloridrato de Raloxifeno/química , Ratos , Moduladores Seletivos de Receptor Estrogênico/químicaRESUMO
The aim of this work was to evaluate the effects of therapeutic doses of Cimicifuga racemosa on cardiovascular parameters and on liver lipid metabolism and redox status in an animal model of estrogen deficiency associated with hypertension, a condition that could make the liver more vulnerable to drug-induced injuries. Female Wistar rats were subjected to the surgical procedures of bilateral ovariectomy (OVX) and induction of renovascular hypertension (two-kidneys, one-clip; 2K1C). These animals (OVX + 2K1C) were treated with daily doses of a C. racemosa extract, using a dose that is similar to that recommended to postmenopausal women (0.6 mg/kg), over a period of 15 days. The results were compared to those of untreated OVX + 2K1C, OVX, and control rats. The treatment with C. racemosa caused a significant reduction in blood pressure. In the liver, treatment did not prevent the development of steatosis, and it reduced the mitochondrial and peroxisomal capacity to oxidize octanoyl-CoA compared to the untreated animals. In addition, C. racemosa caused numerous undesirable effects on the liver redox status: it increased the mitochondrial reactive oxygen species generation, an event that was not accompanied by an increase in the activity of superoxide dismutase, and it induced a decrease in peroxisomal catalase activity. Although the reduced glutathione content had not been affected, a phenomenon that probably reflected the restoration of glucose-6-phosphate dehydrogenase activity by C. racemosa, oxidative damage was evidenced by the elevated level of thiobarbituric acid-reactive substances found in the liver of treated animals.
Assuntos
Anti-Hipertensivos/farmacologia , Cimicifuga/química , Ácidos Graxos/metabolismo , Hipertensão Renovascular/metabolismo , Fígado/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/farmacologia , Acil-CoA Oxidase/metabolismo , Animais , Catalase/metabolismo , Estrogênios/deficiência , Fígado Gorduroso/sangue , Fígado Gorduroso/tratamento farmacológico , Fígado Gorduroso/metabolismo , Feminino , Hipertensão Renovascular/sangue , Hipertensão Renovascular/tratamento farmacológico , Metabolismo dos Lipídeos , Lipídeos/sangue , Fígado/enzimologia , Fígado/metabolismo , Mitocôndrias Hepáticas/metabolismo , Ovariectomia , Oxirredução , Consumo de Oxigênio , Peroxissomos/enzimologia , Peroxissomos/metabolismo , Ratos , Ratos Wistar , Espécies Reativas de Oxigênio/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismoRESUMO
High dietary intake of n-6 fatty acids in relation to n-3 fatty acids may generate health disorders, such as cardiovascular and other chronic diseases. Fish consumption rich in n-3 fatty acids is low in Latin America, it being necessary to seek other alternatives to provide α-linolenic acid (ALA), precursor of n-3 LCPUFA (EPA and DHA). Two innovative oils were assayed, chia (Salvia hispanica) and rosa mosqueta (Rosa rubiginosa). This study evaluated hepatic bioconversion of ALA to EPA and DHA, expression of PPAR-α, acyl-Coenzyme A oxidase 1 (ACOX1) and carnitine acyltransferase I (CAT-I), and accumulation of EPA and DHA in plasma and adipose tissue in Sprague-Dawley rats. Three experimental groups were fed 21 days: sunflower oil (SFO, control); chia oil (CO); rosa mosqueta oil (RMO). Fatty acid composition of total lipids and phospholipids from plasma, hepatic and adipose tissue was assessed by gas-liquid chromatography and TLC. Expression of PPAR-α (RT-PCR) and ACOX1 and CAT-I (Western blot). CO and RMO increased plasma, hepatic and adipose tissue levels of ALA, EPA and DHA and decreased n-6:n-3 ratio compared to SFO (p < 0.05, One-way ANOVA and Newman-Keuls test). CO increased levels of ALA and EPA compared to RMO (p < 0.05). No significant differences were observed for DHA levels. CO also increased the expression of PPAR-α, ACOX1 and CAT-I. Only CAT-I levels were increased by RO. CO and RMO may be a nutritional alternative to provide ALA for its bioconversion to EPA and DHA, and to increase the expression of PPAR-α, ACOX1 and CAT-I, especially CO-oil.
Assuntos
Acil-CoA Oxidase/genética , Carnitina O-Palmitoiltransferase/genética , Ácidos Graxos Ômega-3/metabolismo , PPAR alfa/genética , Óleos de Plantas/metabolismo , Rosa/química , Salvia/química , Ácido alfa-Linolênico/metabolismo , Acil-CoA Oxidase/metabolismo , Animais , Biotransformação , Carnitina O-Palmitoiltransferase/metabolismo , Humanos , PPAR alfa/metabolismo , Óleos de Plantas/administração & dosagem , Ratos , Ratos Sprague-Dawley , Regulação para Cima , Ácido alfa-Linolênico/administração & dosagemRESUMO
Acyl-coenzyme A oxidase 1 (ACOX1) is the first enzyme in peroxisomal fatty acid ß-oxidation; it is rate-limiting and plays a key role in fatty acid metabolism and fat deposition. ACOX1 is an important candidate gene for meat quality selection through marker-assisted selection. Genomic structural analysis showed that bovine ACOX1 shares 86% identity with human ACOX1. Using PCR-SSCP technology, we discovered a single nucleotide polymorphism (SNP) (A1865C) in exon 13 of the ACOX1 gene. Allele frequencies of this SNP were investigated and evaluated with the χ(2) test in 641 cattle populations; only the Jiaxian red population was not in Hardy-Weinberg equilibrium. Gene heterozygosity, effective allele numbers and polymorphism information content of the bovine ACOX1 locus in seven populations varied from 0.2778 to 0.4954, 1.3846 to 1.9817 and 0.2392 to 0.3727, respectively. We also looked for a potential association of this SNP with ultrasound traits in 327 individuals and found a significant effect on ultrasound backfat thickness and ultrasound marbling score (P < 0.05). Meat quality traits were analyzed in another 71 Qinchuan individuals to determine associations with genotype. Animals with genotype AA had higher mean values of backfat thickness than those with genotypes AC and CC. A represents the base before mutation and C represents the base after mutation. We conclude that this SNP of the ACOX1 gene has potential as a genetic marker for meat quality traits in cattle reproduction and breeding.
Assuntos
Acil-CoA Oxidase/genética , Bovinos/genética , Produtos da Carne , Carne , Acil-CoA Oxidase/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Ácidos Graxos/metabolismo , Frequência do Gene , Marcadores Genéticos , Variação Genética , Genótipo , Heterozigoto , Dados de Sequência Molecular , Polimorfismo de Nucleotídeo Único , Característica Quantitativa Herdável , Alinhamento de Sequência , Análise de Sequência de DNA , UltrassonografiaRESUMO
BACKGROUND & AIMS: Bile acid (BA) pool size remains unchanged after cholecystectomy (XGB) but it circulates faster, exposing the enterohepatic system to an increased flux of BA. Triglyceride (TG) and BA metabolisms are functionally inter-related. We investigated whether ablation of the gallbladder (GB) modifies hepatic TG metabolism. METHODS: Male mice were subjected to XGB and fed a normal diet. In some experiments, mice received a 1% nicotinic acid diet to block lipolysis. Parameters of BA and TG metabolism, and microsomal triglyceride transfer protein (MTTP) activity were measured 1-2 months after XGB. Serum parameters, hepatic lipids and mRNA expression of genes of lipid metabolism were determined. RESULTS: BA pool size and synthesis were normal, but biliary BA secretion doubled during the diurnal light phase in XGB mice. Serum and hepatic TG concentrations increased 25% (P<0.02), and hepatic very-low-density lipoproteins (VLDL)-TG and apoB-48 productions increased 15% (P<0.03) and 50% (P<0.01), respectively, after XGB. Feeding a 1% nicotinic acid did normalize VLDL production. MTTP activity increased 15% (P<0.005) after XGB. Hepatic free fatty acid (FFA) synthesis and content, and mRNA levels of lipid metabolism-related genes remained normal in XGD mice. CONCLUSIONS: XGB increased serum and hepatic TG levels, and VLDL production, which were restored to normal by nicotinic acid. The results suggest that FFA flux from adipose tissue to the liver is increased in XGB mice. They support the hypothesis that the GB has a role in the regulation of hepatic TG metabolism and that XGB may favour the accumulation of fat in the liver.
Assuntos
Colecistectomia , Lipoproteínas VLDL/metabolismo , Fígado/metabolismo , Triglicerídeos/metabolismo , Acil-CoA Oxidase/genética , Animais , Apolipoproteína B-100 , Apolipoproteínas B/metabolismo , Ácidos e Sais Biliares/metabolismo , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Colesterol/metabolismo , Citocromo P-450 CYP3A/genética , Ácidos Graxos não Esterificados/metabolismo , Regulação Enzimológica da Expressão Gênica , Lipólise/efeitos dos fármacos , Lipoproteínas VLDL/sangue , Fígado/efeitos dos fármacos , Fígado/enzimologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Niacina/administração & dosagem , RNA Mensageiro/metabolismo , Proteína de Ligação a Elemento Regulador de Esterol 1/genética , Fatores de Tempo , Triglicerídeos/sangue , Regulação para CimaRESUMO
Transgenic tobacco plants capable of over-expressing Xenopus PPARα (xPPARα), a transcription factor known to be required for peroxisome proliferation in animals, were recently generated. These plants (herewith referred to as PPAR-OE) were found to have increased peroxisome abundance, higher peroxisomal acyl-CoA oxidase and catalase activity and modified fatty acid metabolism. Further characterization of PPAR-OE plants revealed a higher susceptibility to virulent and a partial loss of resistance to avirulent Pseudomonas syringae pathogens, whereas the basal resistance response remained unaffected. Biochemical- and defense-related gene expression analyses showed that increased susceptibility to bacterial invasion coincided with the generalized reduction in H(2)O(2) and salicylic acid (SA) levels observed within the first 24 h of bacterial contact. Decreased H(2)O(2) levels were correlated with modified activity levels of catalase and other antioxidant enzymes. A correspondence between a rapid (within 1-24 hpi; ACCO and AOC) and sustained increase (up to 6 days pi; ACCO) in the expression levels of ethylene (ACCO) and jasmonic acid (AOC) biosynthetic genes and a higher susceptibility to virulent bacterial invasion was also observed in PPAR-OE plants. Conversely, no apparent differences in the short- and/or long-term expression levels of markers for the hypersensitive-response, oxidative burst and systemic-acquired resistance were observed between wild type and PPAR-OE plants. The results suggest that peroxisome proliferation could lead to increased susceptibility to bacterial pathogens in tobacco by altering the redox balance of the plant and the expression pattern of key defense signaling pathway genes.
Assuntos
Nicotiana/metabolismo , Nicotiana/microbiologia , PPAR alfa/metabolismo , Peroxissomos/metabolismo , Doenças das Plantas/genética , Pseudomonas syringae/patogenicidade , Acil-CoA Oxidase , Animais , Ascorbato Peroxidases , Biomarcadores/metabolismo , Catalase/metabolismo , Ciclopentanos/análise , Suscetibilidade a Doenças , Regulação da Expressão Gênica de Plantas , Peróxido de Hidrogênio/análise , Oxirredutases/metabolismo , Oxilipinas/análise , PPAR alfa/genética , Peroxidases/metabolismo , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Imunidade Vegetal , Folhas de Planta/metabolismo , Folhas de Planta/microbiologia , Plantas Geneticamente Modificadas/metabolismo , Plantas Geneticamente Modificadas/microbiologia , Ácido Salicílico/análise , Superóxido Dismutase/metabolismo , Fatores de Tempo , Nicotiana/genética , Proteínas de Xenopus/genética , Proteínas de Xenopus/metabolismoRESUMO
AIMS: Maternal diabetes impairs placental development and metabolism. Peroxisome proliferator-activated receptors (PPARs) are ligand-activated nuclear receptors relevant in metabolic homeostasis. We investigated the concentrations of PPARdelta and its endogenous agonist prostacyclin (PGI2), as well as the effects of carbaprostacylin (cPGI(2,) a PPARdelta agonist) on lipid metabolism in placentas from control and streptozotocin-induced diabetic rats on day 13.5 of gestation. MAIN METHODS: The placentas were explanted to evaluate PPARdelta expression and PGI2 concentrations, and cultured with cPGI2 for further analysis of lipid metabolism (concentrations and (14)C-acetate derived synthesis of triglycerides, cholesteryl esters, phospholipids, cholesterol and free fatty acids; release of glycerol and lipid peroxidation). KEY FINDINGS: Reduced PGI2 concentrations were found in the placentas from diabetic rats when compared to controls. cPGI2 additions reduced the concentrations and synthesis of several lipid species, increased lipid catabolism and reduced lipid peroxidation in the placenta. These effects were more marked in diabetic tissues, which presented alterations in the lipid metabolic parameters evaluated. cPGI2 additions increased placental PPARdelta and acyl-CoA oxidase expression, which are changes possibly involved in the catabolic effects observed. SIGNIFICANCE: The present study reveals the capability of cPGI2 to regulate placental lipid metabolism and PPARdelta expression, and suggests that preserving appropriate PGI2 concentrations in the placenta may help to metabolize maternal derived lipid overload in diabetic gestations.
Assuntos
Epoprostenol/análogos & derivados , Lipídeos/análise , PPAR delta/agonistas , Placenta/efeitos dos fármacos , Gravidez em Diabéticas/tratamento farmacológico , Acil-CoA Oxidase/metabolismo , Animais , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/tratamento farmacológico , Epoprostenol/análise , Epoprostenol/farmacologia , Feminino , Glicerol/análise , Metabolismo dos Lipídeos/efeitos dos fármacos , Peroxidação de Lipídeos/efeitos dos fármacos , PPAR delta/análise , Placenta/química , Doenças Placentárias/tratamento farmacológico , Doenças Placentárias/etiologia , Gravidez , Gravidez em Diabéticas/metabolismo , Ratos , Ratos WistarRESUMO
The experiments performed in this report were designed to investigate the mechanisms involved in the metabolic alterations associated with orotic acid-induced hepatic steatosis and the effect of fenofibrate, a stimulant of peroxisome proliferators-activated receptor alpha (PPARalpha), on these alterations. Male Wistar rats were divided into three experimental groups: 1) fed a balanced diet (C); 2) fed a balanced diet supplemented with 1% orotic acid (OA); 3) fed OA diet containing 100 mg.kg(-1) bw.day(-1) fenofibrate (OA+F), for 9 days. Administration of OA to rats induced significant increase in the hepatic total lipids content, marked microvesicular steatosis and decrease in plasma lipids concentrations compared to control group. Fenofibrate treatment prevented fatty liver induction, caused an additional reduction on plasma lipids concentrations and caused a 40% decrease in the lipogenic rate in adipose tissue. The results also showed a 40% increase in lipoprotein lipase (LPL) activity in adipose tissue from OA treated group and fenofibrate administration induced a 50% decrease in LPL activity. The liver mRNA expression of PPARalpha and ACO (acyl CoA oxidase) were 85% and 68% decreased in OA group when compared to control, respectively. Fenofibrate treatment increased the PPARalpha and ACO expressions whereas the CPT-1 (carnitine palmitoyl transferase-1) expression was not altered. Our results have shown that fenofibrate treatment decreases the hepatic lipid content induced by OA which is mediated by an important increase in fatty acid oxidation consequent to an increase in hepatic mRNA expression of PPARalpha and ACO.
Assuntos
Fenofibrato/uso terapêutico , Insuficiência Hepática/induzido quimicamente , Insuficiência Hepática/prevenção & controle , Hipolipemiantes/uso terapêutico , Ácido Orótico/antagonistas & inibidores , Ácido Orótico/toxicidade , Acil-CoA Oxidase/biossíntese , Adipócitos/efeitos dos fármacos , Adipócitos/metabolismo , Agonistas Adrenérgicos beta/farmacologia , Animais , Separação Celular , Dieta , Insuficiência Hepática/patologia , Hipoglicemiantes/farmacologia , Insulina/farmacologia , Isoproterenol/farmacologia , Metabolismo dos Lipídeos/efeitos dos fármacos , Lipídeos/biossíntese , Lipólise/efeitos dos fármacos , Lipase Lipoproteica/metabolismo , Fígado/patologia , Masculino , PPAR alfa/biossíntese , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
We examined the in vivo contribution of insulin, T090137 (T09), agonist of liver X receptor (LXR), fenofibrate, agonist of peroxisome proliferator activated receptor (PPAR-alpha) and sterol regulatory element binding protein-1c (SREBP-1c) on the unsaturated fatty acid synthesis controlled by Delta6 and Delta5 desaturases, compared with the effects on stearoylcoenzyme A desaturase-1. When possible they were checked at three levels: messenger RNA (mRNA), desaturase protein and enzymatic activity. In control rats, only fenofibrate increased the insulinemia that was maintained by the simultaneous administration of T09, but this increase has no specific effect on desaturase activity. T09 enhanced SREBP-1 in control animals and the mRNAs and activity of the three desaturases in control and type-1 diabetic rats, demonstrating a LXR/SREBP-1-mediated activation independent of insulin. However, simultaneous administration of insulin and T09 to diabetic rats led to a several-fold increase of the mRNAs of the desaturases, suggesting a strong synergic effect between insulin and LXR/retinoic X receptor (RXR). Moreover, this demonstrates the existence of an interaction between unsaturated fatty acids and cholesterol metabolism performed by the insulin/SREBP-1c system and LXR/RXR. PPAR-alpha also increased the expression and activity of the three desaturases independently of the insulinemia since it was equivalently evoked in streptozotocin diabetic rats. Besides, PPAR-alpha increased the palmitoylcoenzyme A elongase, evidencing a dual regulation in the fatty acid biosynthesis at the level of desaturases and elongases. The simultaneous administration of fenofibrate and T09 did not show additive effects on the mRNA expression and activity of the desaturases. Therefore, the results indicate a necessary sophisticated interaction of all these factors to produce the physiological effects.
Assuntos
Proteínas de Ligação a DNA/fisiologia , Ácidos Graxos Dessaturases/metabolismo , Ácidos Graxos Insaturados/biossíntese , Insulina/fisiologia , PPAR alfa/fisiologia , Receptores Citoplasmáticos e Nucleares/fisiologia , Acil-CoA Oxidase/genética , Acil-CoA Oxidase/metabolismo , Animais , Sequência de Bases , Colesterol/sangue , Primers do DNA , Ativação Enzimática , Ácidos Graxos Dessaturases/genética , Fenofibrato/farmacologia , Insulina/sangue , Receptores X do Fígado , Masculino , Receptores Nucleares Órfãos , RNA Mensageiro/genética , Ratos , Ratos Wistar , Proteína de Ligação a Elemento Regulador de Esterol 1/metabolismoRESUMO
Beauveria bassiana produces acyl-Co oxidase (ACO) in the P(20000 g) fraction of glucose and alkane-grown cultures that catalyze the oxidation of acyl-CoAs of different chain length. The activity was measured indirectly over the formation of H2O2 via the oxidative-coupled assay system. ACO activity was assessed spectrophotometrically in the P(20000 g) fraction of glucose-grown (FS0) and n-alkane grown cultures (FS(alk)), employing acyl-CoAs of 16 to 24 carbons as substrates. A significant increment in the activity was observed in FS(alk) as compared to that of controls (FS0) in all conditions tested. Tetracosane-grown cultures showed the highest activity with lignoceroyl-CoA. The reaction conditions were optimized employing lignoceroyl-CoA as substrate. A variable lag phase was observed when the activity was measured as a function of time. In the presence of 3-amino-1,2,4-triazole (AT) to prevent H2O2 consumption by endogenous catalase, the lag phase became shorter and disappeared when AT concentrations were raised from 40 to 200 mM, thus enhancing acyl-CoA oxidation. Enzyme activity reached its maximal value in the presence of 240 microg peroxidase, 0.08% Triton X-100 and 36 microM bovine serum albumin. The apparent Km using lignoceroyl as substrate was estimated 2.5 microM. ACO showed high activity and stability between 30 and 40 degrees C, as well as between 7.0 and 9.0 pH, for 120 min, being 7.0 the optimum pH.
Assuntos
Acil-CoA Oxidase/metabolismo , Beauveria/enzimologia , Acil-CoA Oxidase/antagonistas & inibidores , Alcanos/metabolismo , Amitrol (Herbicida)/farmacologia , Beauveria/metabolismo , Inibidores Enzimáticos/farmacologia , Concentração de Íons de Hidrogênio , Cinética , Espectrofotometria UltravioletaRESUMO
Both insulin and PPAR-alpha up-modulate hepatic Delta9, Delta6 and Delta5 desaturating enzymes involved in the biosynthesis of mono- and polyunsaturated fatty acids. Currently, we have examined for 9 days the independent and simultaneous effects of daily glargine insulin and fenofibrate administration on the insulinemia, glycemia, hepatic acyl-CoA oxidase activity and mRNAs and enzymatic activities of stearoyl-CoA desaturase-1 (SCD-1) and Delta5 desaturase in streptozotocin diabetic rats. Glargine insulin depressed the hyperglycemia of diabetic rats at 4h, but not after 24h of injection. Fenofibrate increased the radioimmunoreactive insulinemia in non-diabetic rats without changing the glycemia. Insulin increased the mRNAs and activities of SCD-1 and Delta5 desaturase depressed in diabetic rats. Fenofibrate increased acyl-CoA oxidase activity, and the mRNAs and activities of both desaturating enzymes in non-diabetic, diabetic and insulin-treated diabetic rats, but was less effective in the mRNAs modification of diabetic animals. Therefore, insulin, and fenofibrate through PPAR-alpha activation, enhance liver mRNAs and activities of SCD-1 and Delta5 desaturases independently and synergistically through different mechanisms. Insulin and fenofibrate independently increased the 18:1/18:0 ratio in liver lipids, increasing the fluidity of the membranes. The 20:4/18:2 ratio was maintained. Fenofibrate increased palmitic acid, but decreased stearic acid percentage in liver lipids.
Assuntos
Diabetes Mellitus Experimental/sangue , Ácidos Graxos Insaturados/biossíntese , Fenofibrato/administração & dosagem , Insulina/administração & dosagem , Insulina/sangue , Acil-CoA Oxidase/efeitos dos fármacos , Acil-CoA Oxidase/metabolismo , Animais , Glicemia/efeitos dos fármacos , Glicemia/metabolismo , Dessaturase de Ácido Graxo Delta-5 , Diabetes Mellitus Experimental/induzido quimicamente , Diabetes Mellitus Experimental/enzimologia , Modelos Animais de Doenças , Ácidos Graxos Dessaturases/efeitos dos fármacos , Ácidos Graxos Dessaturases/metabolismo , Insulina/análogos & derivados , Lipídeos/química , Fígado/química , Fígado/enzimologia , Fígado/metabolismo , Masculino , RNA Mensageiro/efeitos dos fármacos , Ratos , Ratos Wistar , Estearoil-CoA Dessaturase/efeitos dos fármacos , Estearoil-CoA Dessaturase/metabolismo , EstreptozocinaRESUMO
The fomesafen and 2,4-D amine herbicide induce cytotoxic effects at hepatic level in rats, such as: hepatomegaly, hyperplasia and increase in the enzymes activity which participate in the processes of peroxisomal beta-oxidation of fatty acids. In this work, the effect of vitamin E and C was evaluated, as well as, the dexamethasone in the modulation of these hepatotoxic effects. Sprague-Dawley rats were treated with the herbicides and with the agents to be evaluated. The different treatments were given during 15 days orally route. The herbicides combined with the dexamethasone and antioxidant agents were administrated only and simultaneously with the herbicides. Once concluded the different treatment, the rats were weighed and sacrificed. It was evaluated the liver size and liver fragments were obtained to determine the enzymatic activity of Fatty Acyl CoA-oxidase (FACO) and cellular number. The results showed that the hepatomegaly induced by fomesafen was inhibited by the vitamins and by the dexamethasone, while any effect was not observed in the group of rats treated with 2,4-D amine. None of the agents modulated the FACO activity induced by herbicides in treated rats. However, the dexamethasone showed a protective effect in the hyperplasia induced by two herbicides. The hepatotoxic effects induced by the herbicides responded to a different mechanism due to the differences of the effects observed at the antioxidant agents. On the other hand, the inhibition of the cellular proliferation by the dexamethasone does not keep relation with the responsible mechanisms of inducing the oxidant stress into FACO activity. Under experimental conditions of this study, the use of these agents does not guarantee protection against the hepatotoxic effects induced by the herbicides.
Assuntos
Ácido 2,4-Diclorofenoxiacético/antagonistas & inibidores , Antioxidantes/farmacologia , Benzamidas/antagonistas & inibidores , Doença Hepática Induzida por Substâncias e Drogas , Dexametasona/farmacologia , Dimetilaminas/antagonistas & inibidores , Herbicidas/antagonistas & inibidores , Vitaminas/farmacologia , Ácido 2,4-Diclorofenoxiacético/toxicidade , Acil-CoA Oxidase , Animais , Ácido Ascórbico/farmacologia , Benzamidas/toxicidade , Dimetilaminas/toxicidade , Hepatomegalia/induzido quimicamente , Herbicidas/toxicidade , Hiperplasia/induzido quimicamente , Fígado/efeitos dos fármacos , Fígado/enzimologia , Masculino , Oxirredutases/efeitos dos fármacos , Oxirredutases/metabolismo , Ratos , Ratos Sprague-Dawley , Vitamina E/farmacologiaRESUMO
Fenofibrate, the hypolipidemic drug and peroxisome proliferator, was given to mice (0.23% w/w in the diet) during 1-3 weeks and enzyme activities, H2O2 concentration, and H2O2 production rate were determined. A maximal increase of 150% in liver/body weight ratio was observed after 3 weeks of treatment. Acyl-CoA oxidase, catalase and uricase activities were increased by 712%, 506% and 41% respectively by treatment with fenofibrate. Se- and non Se-glutathione peroxidase and Mn-superoxide dismutase activities were increased by 331%, 188% and 130% respectively in the liver of 2 weeks-treated mice. Cu-Zn superoxide dismutase activity was not affected by fenofibrate treatment. H2O2 steady-state concentration showed an increase of 89% after 2 weeks of treatment. H2O2 production rates, and the steady-state concentrations of the intermediates HO, R and ROO, calculated using experimental data, were higher in the liver of fenofibrate-treated mice than in control animals. According to our findings, the imbalance between H2O2 production and its degradation by its metabolizing enzymes during peroxisome proliferation, would result in an increased level of H2O2 steady-state concentration, with the resulting oxidative stress which may lead to the generation of oxidative damage and to the induction of liver carcinogenesis.
Assuntos
Fenofibrato/farmacologia , Peróxido de Hidrogênio/metabolismo , Hipolipemiantes/farmacologia , Fígado/metabolismo , Microcorpos/metabolismo , Acil-CoA Oxidase , Animais , Antioxidantes/metabolismo , Antioxidantes/farmacologia , Peso Corporal , Catalase/metabolismo , Dieta , Feminino , Fenofibrato/administração & dosagem , Glutationa Peroxidase/metabolismo , Cinética , Peroxidação de Lipídeos/efeitos dos fármacos , Fígado/efeitos dos fármacos , Camundongos , Microcorpos/efeitos dos fármacos , Oxirredutases/metabolismo , Superóxido Dismutase/metabolismo , Urato Oxidase/metabolismoRESUMO
Immunohistochemical studies with antisera against four peroxisomal enzymes, catalase and beta-oxidation enzymes (acyl-coenzyme A oxidase, bifunctional protein, and 3-ketoacyl-CoA thiolase), were performed on brain, liver, and kidney specimens from patients with peroxisomal disorders, as well as specimens from three control subjects, by using conventional paraffin-embedded autopsy material. The patients included eight with Zellweger syndrome and one with neonatal adrenoleukodystrophy. In the liver and kidney specimens from all patients, except one with Zellweger syndrome, diffuse immunostaining with all antisera in the cytoplasm of hepatocytes and renal tubular epithelium suggested an absence of peroxisomes but the presence of peroxisomal enzymes. Examination of brain specimens indicated a weak or negative reaction of neurons in the cerebral cortex and a weak reaction of glial cells in the white matter, which suggested maturational delay compared with control subjects. The delayed immunoreactive pattern of peroxisomal enzymes in Zellweger syndrome and neonatal adrenoleukodystrophy may be related to the significant neuropathologic features of polymicrogyria and dysmyelinogenesis. One patient with Zellweger syndrome had a unique finding of a positive granular catalase reaction and a negative reaction with antisera to 3-ketoacyl-coenzyme A thiolase, which suggested a diagnosis of pseudo-Zellweger syndrome. This study validates the application of these immunohistochemical methods to the study of peroxisomal enzymes. Use of these methods improves the accuracy of diagnosis of peroxisomal disorders.
Assuntos
Adrenoleucodistrofia/diagnóstico , Encéfalo/patologia , Rim/patologia , Fígado/patologia , Microcorpos/enzimologia , Síndrome de Zellweger/diagnóstico , Acetil-CoA C-Aciltransferase/análise , Acil-CoA Oxidase , Catalase/análise , Pré-Escolar , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Neuroglia/patologia , Neurônios/patologia , Oxirredutases/análiseRESUMO
We describe an infant girl with a clinical, chemical, and pathologic syndrome remarkably similar to Zellweger cerebrohepatorenal syndrome but whose liver parenchymal cells contained abundant peroxisomes. Peroxisomal L-alpha hydroxy acid oxidase, catalase, and the plasmalogen synthesizing enzyme dihydroxy acetone phosphate-acyl transferase activities were normal; other peroxisomal enzymatic activities, including fatty acyl-CoA oxidase and D-amino acid oxidase, were reduced by 80% to 85%. Oxidation of bile acids and pipecolic acid was also deficient. Autopsy revealed the presence of neuronal heterotopia, renal cortical cysts, adrenal atrophy, and accumulation of very long chain fatty acids. The clinical and pathologic features of this case of "pseudo-Zellweger syndrome" reflect a deficiency in multiple peroxisomal activities rather than a defect in peroxisomal biogenesis. The deficient enzymatic activities require flavin adenine dinucleotide, and the underlying defect may be in the utilization of this cofactor.