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1.
Food Funct ; 15(11): 6147-6163, 2024 Jun 04.
Artigo em Inglês | MEDLINE | ID: mdl-38767501

RESUMO

Scope: fructose consumption from added sugars correlates with the epidemic rise in MetS and CVD. Maternal fructose intake has been described to program metabolic diseases in progeny. However, consumption of fructose-containing beverages is allowed during gestation. Cholesterol is also a well-known risk factor for CVD. Therefore, it is essential to study Western diets which combine fructose and cholesterol and how maternal fructose can influence the response of progeny to these diets. Methods and results: a high-cholesterol (2%) diet combined with liquid fructose (10%), as a model of an unhealthy Western diet, was administered to descendants from control and fructose-fed mothers. Gene (mRNA and protein) expression and plasma, fecal and tissue parameters of cholesterol metabolism were measured. Interestingly, progeny from fructose-fed dams consumed less liquid fructose and cholesterol-rich chow than males from control mothers. Moreover, descendants of fructose-fed mothers fed a Western diet showed an increased cholesterol elimination through bile and feces than males from control mothers. Despite these mitigating circumstances to develop a proatherogenic profile, the same degree of hypercholesterolemia and severity of steatosis were observed in all descendants fed a Western diet, independently of maternal intake. An increased intestinal absorption of cholesterol, synthesis, esterification, and assembly into lipoprotein found in males from fructose-fed dams consuming a Western diet could be the cause. Moreover, an augmented GLP2 signalling seen in these animals would explain this enhanced lipid absorption. Conclusions: maternal fructose intake, through a fetal programming, makes a Western diet considerably more harmful in their descendants than in the offspring from control mothers.


Assuntos
Colesterol , Dieta Ocidental , Frutose , Animais , Frutose/efeitos adversos , Frutose/administração & dosagem , Feminino , Masculino , Ratos , Dieta Ocidental/efeitos adversos , Gravidez , Colesterol/metabolismo , Colesterol/sangue , Efeitos Tardios da Exposição Pré-Natal , Ratos Wistar , Fenômenos Fisiológicos da Nutrição Materna , Fígado/metabolismo , Hipercolesterolemia/metabolismo , Hipercolesterolemia/etiologia
2.
Artigo em Inglês | MEDLINE | ID: mdl-19594420

RESUMO

Peroxisome proliferator-activated receptor alpha (PPARalpha) regulates transcription of genes involved both in lipid and glucose metabolism as well as in inflammation. Fibrates are PPARalpha ligands used to normalize lipid and glucose parameters and exert antiinflammatory effects. In fact, fibrates have already been demonstrated to benefit metabolic syndrome, type 2 diabetes and cardiovascular diseases. This article reviews the mechanism of action and the functional roles of fibrates, emphasizing the factors modulating their capacity to activate PPARalpha and affecting their effectiveness. These factors may possibly explain the findings obtained in animal studies and clinical trials with fibrates which showed either untoward effects and/or inefficient hypolipidemic action of PPARalpha activation. We also discuss briefly the natural and synthetic agonists of PPARalphawhich are currently being developed and supposedly display greater effectiveness and fewer adverse effects than fibrates.


Assuntos
Ácido Clofíbrico/uso terapêutico , PPAR alfa/agonistas , Animais , Ácido Clofíbrico/farmacologia , Regulação da Expressão Gênica , Glucose/metabolismo , Humanos , Inflamação/etiologia , Metabolismo dos Lipídeos , PPAR alfa/análise , PPAR alfa/fisiologia , Processamento de Proteína Pós-Traducional , Transdução de Sinais
3.
J Physiol Biochem ; 62(3): 189-98, 2006 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-17451160

RESUMO

The present study was addressed to determine whether the high expression of peroxisome proliferator-activated receptor-alpha (PPAR-alpha) in rat liver during the perinatal stage plays a role in the induction of liver lipoprotein lipase (LPL) expression and activity. Parallel increases in liver mRNA PPAR-alpha and LPL activity were found in newborn rats, and after a slight decline, values remained elevated until weaning. Anticipated weaning for 3 days caused a decline in those two variables as well as in the mRNA LPL level, and a similar change was also found in liver triacylglycerol concentration. Force-feeding with Intralipid in 10-day-old rats or animals kept fasted for 5 h showed high mRNA-PPARalpha and -LPL levels as well as LPL activity with low plasma insulin and high FFA levels, whereas glucose and a combination of glucose and Intralipid produced low mRNA-PPARalpha and -LPL levels as well as LPL activity. Under these latter conditions, plasma insulin and FFA levels were high in those rats receiving the combination of glucose and Intralipid, whereas plasma FFA levels were low in those force-fed with glucose. It is proposed that the hormonal and nutritional induction of liver PPAR-alpha expression around birth and its maintained elevated level throughout suckling is responsible for the induction of liver LPL-expression and activity during suckling.


Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Regulação Enzimológica da Expressão Gênica , Lipase Lipoproteica/genética , Fígado , PPAR alfa/genética , Animais , Animais Recém-Nascidos , Animais Lactentes , Emulsões Gordurosas Intravenosas/farmacologia , Ácidos Graxos/sangue , Feminino , Glucose/farmacologia , Insulina/sangue , Fígado/embriologia , Fígado/crescimento & desenvolvimento , Fígado/fisiologia , Masculino , Gravidez , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Triglicerídeos/metabolismo
4.
Oncology ; 62(3): 286-90, 2002.
Artigo em Inglês | MEDLINE | ID: mdl-12065877

RESUMO

OBJECTIVES: The uPA-PAI system has been shown to play a role in the development of a more aggressive tumor phenotype. The PAI-1 promoter 4G/5G polymorphism, furthermore, regulates free plasma PAI-1 levels in patients with myocardial infarction. Our aim was to verify if the different polymorphisms in the PAI-1 promoter are also associated with alterations in the intracellular accumulation of uPA-PAI complexes in human breast cancer. METHODS: Accumulation of uPA-PAI complexes inside the tumor cells was determined by means of immunohistochemistry, as previously described by our own group, and two extremely different sets of tumors were chosen, one of them with strong uPA-PAI complex reactivity inside more than 50% of tumor cells, the other with no demonstrable reactivity at all. Finally, the 4G/5G polymorphism of the PAI-1 promoter was studied in all of them by means of DNA extraction, PCR amplification of the PAI promoter sequence, and restriction polymorphism typing. RESULTS: Absence of intracellular uPA-PAI complex accumulation was significantly associated with the prevalence of the 4G allele and, conversely, the presence of uPA-PAI complexes inside the tumor cells was significantly associated with 5G/5G homozygosity (logistic regression, p = 0.0128). Furthermore, none of the 7 5G/5G homozygous tumors showed histological grade 3, as did 6/21 tumors in the group where the 4G allele was present. In spite of the low case number, this association of the 5G/5G polymorphism with a less aggressive phenotype almost reached statistical significance (Spearman's correlation test, p = 0.118). CONCLUSIONS: The 4G/5G polymorphism of the PAI-1 promoter seems indeed to be associated with different rates of uPA-PAI complex internalization by breast cancer cells. Complex accumulation inside the tumor cells is significantly related to 5G/5G homozygosity, and this shows a trend towards an association with a less aggressive, better-differentiated tumor phenotype.


Assuntos
Neoplasias da Mama/genética , Inibidor 1 de Ativador de Plasminogênio/genética , Inibidor 1 de Ativador de Plasminogênio/metabolismo , Polimorfismo Genético , Ativador de Plasminogênio Tipo Uroquinase/metabolismo , Neoplasias da Mama/enzimologia , Primers do DNA/química , DNA de Neoplasias/metabolismo , Feminino , Genótipo , Homozigoto , Humanos , Técnicas Imunoenzimáticas , Ativadores de Plasminogênio/metabolismo , Reação em Cadeia da Polimerase , Regiões Promotoras Genéticas , Receptores de Estrogênio/análise , Receptores de Progesterona/análise
5.
Arch Biochem Biophys ; 394(2): 182-8, 2001 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-11594732

RESUMO

It was previously found that the expression of peroxisome proliferator-activated receptor-alpha (PPARalpha) was markedly augmented in the liver of suckling rats, in comparison to the fetuses and most notably to adult rats and it paralleled similar changes in hepatic lipid concentration. To determine whether these changes could be related to the high lipid content of the maternal milk and/or to hormonal status, the role of changes in nutrient availability and in plasma insulin concentration on liver expression during the perinatal stage in vivo in the rat was studied. When suckling rats were weaned on day 17, instead of on day 20, the level of hepatic PPARalpha mRNA decreased earlier than in rats weaned later. When 10-day-old rats were force-fed with either glucose or Intralipid or a combination of both diets, it was found that, at similar low levels of plasma insulin, a high level of FFA stimulated PPARalpha expression, whereas, at similar high plasma FFA concentrations, an elevated insulin level attenuated the increase in PPARalpha expression. It is proposed that both the high lipid intake and decreased plasma insulin level are responsible for the high PPARalpha expression detected in rat neonates.


Assuntos
Fenômenos Fisiológicos da Nutrição Animal , Regulação da Expressão Gênica , Insulina/sangue , Fígado/metabolismo , Receptores Citoplasmáticos e Nucleares/metabolismo , Fatores de Transcrição/metabolismo , Acil-CoA Oxidase , Administração Oral , Envelhecimento/metabolismo , Animais , Animais Lactentes , Dieta , Emulsões Gordurosas Intravenosas/administração & dosagem , Emulsões Gordurosas Intravenosas/metabolismo , Ácidos Graxos não Esterificados/sangue , Feminino , Glucose/administração & dosagem , Glucose/metabolismo , Metabolismo dos Lipídeos , Lipídeos/análise , Fígado/química , Oxirredutases/genética , Oxirredutases/metabolismo , RNA Mensageiro/análise , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores Citoplasmáticos e Nucleares/genética , Fatores de Transcrição/genética , Desmame
6.
Biochimie ; 82(8): 723-6, 2000 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-11018288

RESUMO

The expression of the peroxisome proliferator-activated receptor-alpha (PPARalpha) as well as of some related genes was studied in rat liver at different stages of development (from 19-day-old fetuses to 1 month-old rats). The level of PPARalpha mRNA appeared higher in neonates than in fetuses or 1 month-old rats. Whereas the pattern for phosphoenolpyruvate carboxykinase (PEPCK) mRNA level was similar to that of PPARalpha, the mRNA level of both acyl-CoA oxidase (ACO) and apolipoprotein CIII (apo CIII) showed diverse profiles. Western blotting analysis also revealed an increased level of PPARalpha protein in liver of suckling rats. Similarities of mRNA PEPCK and PPARalpha expression indicate a common control mechanism, where both nutritional and hormonal factors may be involved.


Assuntos
Desenvolvimento Embrionário e Fetal , Regulação da Expressão Gênica no Desenvolvimento , Fígado/metabolismo , Receptores Citoplasmáticos e Nucleares/genética , Fatores de Transcrição/genética , Transcrição Gênica , Acil-CoA Oxidase , Envelhecimento , Animais , Animais Recém-Nascidos , Apolipoproteína C-III , Apolipoproteínas C/genética , Fígado/embriologia , Fígado/crescimento & desenvolvimento , Proteínas Nucleares/genética , Oxirredutases/genética , Fosfoenolpiruvato Carboxiquinase (GTP)/genética , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley
7.
Mol Pharmacol ; 54(3): 463-73, 1998 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-9730905

RESUMO

In this study, we show that peroxisome proliferator chemical (PPC) exposure leads to alterations in the expression of genes in rat liver regulated by the sex-specific growth hormone secretory pattern and induced during inflammation. Expression of the male-specific cytochrome P450 (P450) 2C11 and alpha2 urinary globulin (alpha2u) genes and the female-specific P450 2C12 gene was down-regulated by some PPC. Expression of P450 2C13, also under control by the sex-specific growth hormone secretory pattern, was not altered by PPC treatment, indicating that regulation of CYP2C family members does not involve perturbation of the growth hormone secretory pattern. In contrast to the increases in expression observed when inflammation was induced in male rats, two positive acute-phase response genes, alpha1-acid glycoprotein and beta-fibrinogen, were decreased by PPC exposure. The down-regulation of the P450 2C11 by WY-14,643 could be reproduced in cultured rat hepatocytes, indicating the down-regulation is a direct effect. Experiments in wild-type mice and mice that lacked a functional peroxisome proliferator-activated receptor-alpha gene showed that down-regulation by WY of alpha1-acid glycoprotein, beta-fibrinogen, and a mouse homologue of alpha2u was dependent on peroxisome proliferator-activated receptor-alpha expression. Our results demonstrate that PPC exposure leads to down-regulation of diverse liver-specific genes, including CYP2C family members important in hormonal homeostasis and acute-phase response genes important in inflammatory responses.


Assuntos
Reação de Fase Aguda/enzimologia , Reação de Fase Aguda/genética , Hidrocarboneto de Aril Hidroxilases , Sistema Enzimático do Citocromo P-450/biossíntese , Sistema Enzimático do Citocromo P-450/genética , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Fígado/fisiologia , Microcorpos/efeitos dos fármacos , Esteroide 16-alfa-Hidroxilase , Esteroide Hidroxilases/biossíntese , Esteroide Hidroxilases/genética , Proteínas de Fase Aguda/metabolismo , alfa-Globulinas/biossíntese , alfa-Globulinas/genética , Animais , Células Cultivadas , Regulação para Baixo/efeitos dos fármacos , Feminino , Fígado/efeitos dos fármacos , Fígado/enzimologia , Masculino , Camundongos , Ratos , Ratos Endogâmicos F344 , Ratos Sprague-Dawley , Receptores Citoplasmáticos e Nucleares/fisiologia , Fatores de Transcrição/fisiologia
8.
Environ Toxicol Pharmacol ; 6(1): 59-66, 1998 Aug 04.
Artigo em Inglês | MEDLINE | ID: mdl-21781882

RESUMO

In vitro effects of pantethine on adipose tissue lipolysis and on both hepatic and intestinal cholesterol and fatty acid synthesis in normolipidemic rats are determined and related to their respective in vivo hypolipidemic effects after acute oral administration. At 3, 5, 7 and 24 h after a single high dose of pantethine to rats, free fatty acids (FFA), cholesterol and triglycerides levels decreased whereas plasma glycerol increased, the effect becoming significant at 7 h. The release of glycerol and FFA by epididymal fat pad pieces from rats was measured in Krebs Ringer bicarbonate-albumin buffer supplemented or not with epinephrine and several concentrations of pantethine (0, 10(-5), 10(-4), or 10(-3) M), and it turned out to be enhanced as pantethine concentration increased. Besides, when glucose was present in the medium, this drug lowered fatty acid re-esterification in a dose-dependent manner, the effect being specially evident in the presence of epinephrine. In vitro synthesis of both cholesterol and fatty acids by slices of liver or intestinal epithelial cells was depressed as the concentration of pantethine increased in the medium. Thus, an inhibition of both cholesterolgenesis and lipogenesis seems to contribute to the hypocholesterolemic and hypotriglyceridemic effects of pantethine. On the other hand, the stimulation of lipolysis and the inhibition of fatty acid re-esterification on adipose tissue caused by pantethine must be counteracted by a high fatty acid oxidation in the liver which would explain the decrease in FFA and the increase in glycerol levels detected in the plasma of the pantethine-treated animals.

9.
Cancer Lett ; 134(1): 61-71, 1998 Dec 11.
Artigo em Inglês | MEDLINE | ID: mdl-10381131

RESUMO

To better understand the molecular basis of the hepatocyte proliferation and induction of hepatocellular adenomas by exposure to peroxisome proliferator chemicals (PPC), a systematic search for genes modulated by a PPC (WY-14643) in rat liver was carried out using the differential display technique. The fragments fell into two classes based on the time of initial and maximal induction by WY-14643. The class I genes (clones 5 and 30) were induced 3 h after a gavage exposure to WY-14643 with maximal expression at 24 h. The class II genes (clones 13 and 16) were induced after 24 h with maximal expression at 78 weeks. Expression of the class II genes was also increased after other treatments that cause cell proliferation. Clone 30 was identified as CYP4A2, previously shown to be regulated by PPC. Clone 13 was homologous to the mouse protein H gene, a component of the heterogeneous nuclear ribonucleoprotein particle important in mRNA splicing. Clone 16 was identified as cyclophilin-A, the receptor for the immunosuppressant drug cyclosporin A. The sequence of clone 5 was unique. These data demonstrate that WY-14643 increases the levels of a number of novel genes that are coordinately regulated with increases in chronic cell proliferation and fatty acid metabolism.


Assuntos
Genes/genética , Neoplasias Hepáticas Experimentais/genética , Proliferadores de Peroxissomos/efeitos adversos , Pirimidinas/efeitos adversos , Animais , Sequência de Bases , Clonagem Molecular , Citocromo P-450 CYP4A , Sistema Enzimático do Citocromo P-450/genética , DNA Complementar/química , DNA Complementar/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia , Neoplasias Hepáticas Experimentais/induzido quimicamente , Masculino , Oxigenases de Função Mista/genética , Dados de Sequência Molecular , Peptidilprolil Isomerase/genética , Ratos , Ratos Endogâmicos F344 , Análise de Sequência de DNA , Fatores de Tempo
10.
Biochimie ; 79(2-3): 151-62, 1997.
Artigo em Inglês | MEDLINE | ID: mdl-9209713

RESUMO

Exposure to some peroxisome proliferator chemicals (PPC) leads to toxic effects on sex organ function possibly by alterations of steroid hormone metabolism. A systematic search for genes whose mRNA levels are modulated by the PPC WY-14643 (WY) was carried out in rat liver, a site of steroid hormone metabolism. The sequence of one up-regulated cDNA (2480 bp) was predicted to encode a protein of 735 amino acids with 82% identity to the porcine 17 beta-hydroxysteroid dehydrogenase type IV (HSD IV) originally isolated as a 17 beta-estradiol dehydrogenase. The rat HSD IV was localized to peroxisomes and was regulated by diverse PPC by two distinct mechanisms. Induction of HSD IV and acyl-CoA oxidase (ACO) proteins in rat liver at different treatment times and concentrations of gemfibrozil (GEM) and di-n-butyl phthalate (DBP) were almost identical, suggesting that HSD IV mRNA induction involves the peroxisome proliferator-activated receptor alpha, a regulator of ACO. In contrast, HSD IV protein levels were only weakly induced by WY, a strong inducer of ACO protein, even though the levels of both HSD IV and ACO mRNA were strongly stimulated by WY. Thus HSD IV protein levels were uniquely regulated pretranslationally by WY. In addition to HSD IV we also identified the male-specific alpha 2u-globulin as a PPC down-regulated gene. This prompted us to examine the expression of another male-specific gene, CYP2C11, that catalyzes the hydroxylations of estradiol at the 2 and 16 alpha positions. Cyp2C11 protein expression in rat liver was either decreased or completely abolished after a 3-week treatment by GEM or WY, respectively. Decreased expression of enzymes which inactivate estradiol including Cyp2C11, and the reported increased expression of aromatase may explain why male rats exposed to diverse PPC have higher serum estradiol levels. These higher estradiol levels in male rats have been thought to be mechanistically linked to Leydig cell hyperplasia and adenomas. Increased conversion of estradiol to the less active estrone by HSD IV induction may explain how exposure to the phthalate di-(2-ethylhexyl) phthalate leads to decreases in serum estradiol levels and suppression of ovulation in female rats.


Assuntos
17-Hidroxiesteroide Desidrogenases/genética , Hidrocarboneto de Aril Hidroxilases , Enoil-CoA Hidratase , Estrogênios/metabolismo , Microcorpos/efeitos dos fármacos , Complexos Multienzimáticos , Esteroide 16-alfa-Hidroxilase , Sequência de Aminoácidos , Animais , Sequência de Bases , Compartimento Celular , Clonagem Molecular , Sistema Enzimático do Citocromo P-450/genética , Família 2 do Citocromo P450 , Regulação para Baixo , Estradiol/metabolismo , Feminino , Genfibrozila/farmacologia , Expressão Gênica , Hipolipemiantes/farmacologia , Masculino , Microcorpos/metabolismo , Dados de Sequência Molecular , Proteína Multifuncional do Peroxissomo-2 , Pirimidinas/farmacologia , RNA Mensageiro/genética , Ratos , Ratos Endogâmicos F344 , Ratos Sprague-Dawley , Esteroide Hidroxilases/genética
11.
Mol Pharmacol ; 50(5): 1157-66, 1996 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-8913347

RESUMO

To better understand the molecular mechanisms of the pleiotropic responses induced by exposure to peroxisome proliferator chemicals (PPCs), we conducted a systematic search for genes whose mRNA levels are modulated by the PPC WY-14,643 (WY) in rat liver. The sequence of one up-regulated cDNA (2480 bp) was predicted to encode a protein of 735 aa with 82% identity to the porcine 17 beta-hydroxysteroid dehydrogenase type IV (HSD IV). Like the porcine enzyme, the rat HSD IV contains' a region homologous to yeast hydratase-dehydrogenase-epimerases and to sterol carrier proteins, indicating that the rat HSD IV has broad substrate specificity and contributes to cholesterol metabolism. The rat HSD IV was regulated by diverse PPCs via two distinct mechanisms. Induction of HSD IV and acyl-CoA oxidase (ACO) proteins in rat liver at different treatment times and concentrations of gemfibrozil and di-n-butyl phthalate were almost identical, indicating that HSD IV mRNA induction involves the peroxisome proliferator-activated receptor alpha, a regulator of ACO. In contrast, HSD IV protein levels were only weakly induced by WY, a strong inducer of ACO protein, even though the levels of HSD IV and ACO mRNA were strongly stimulated by WY and gemfibrozil. Thus, HSD IV protein levels were uniquely regulated pretranslationally by WY via a novel mechanism. Increased conversion of estradiol to the less-active estrone by HSD IV induction may explain how phthalate exposure leads to decreases in serum estradiol levels and suppression of ovulation.


Assuntos
17-Hidroxiesteroide Desidrogenases/genética , Anticolesterolemiantes/farmacologia , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Isoenzimas/genética , Microcorpos/fisiologia , Pirimidinas/farmacologia , 17-Hidroxiesteroide Desidrogenases/biossíntese , 17-Hidroxiesteroide Desidrogenases/classificação , Sequência de Aminoácidos , Animais , Sequência de Bases , DNA Complementar/genética , DNA Complementar/isolamento & purificação , DNA Complementar/metabolismo , Indução Enzimática/efeitos dos fármacos , Estradiol/metabolismo , Regulação Enzimológica da Expressão Gênica/fisiologia , Isoenzimas/biossíntese , Fígado/efeitos dos fármacos , Fígado/enzimologia , Fígado/fisiologia , Masculino , Microcorpos/efeitos dos fármacos , Microcorpos/enzimologia , Dados de Sequência Molecular , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Endogâmicos F344 , Ratos Sprague-Dawley , Receptores Citoplasmáticos e Nucleares/efeitos dos fármacos , Receptores Citoplasmáticos e Nucleares/fisiologia , Fatores de Transcrição/efeitos dos fármacos , Fatores de Transcrição/fisiologia , Regulação para Cima/efeitos dos fármacos
12.
Environ Toxicol Pharmacol ; 2(4): 351-7, 1996 Dec 20.
Artigo em Inglês | MEDLINE | ID: mdl-21781742

RESUMO

The release of both glycerol and free fatty acids (FFA) into a medium by epididymal fat pad pieces from fed rats incubated in Krebs Ringer bicarbonate-albumin buffer supplemented or not with epinephrine decreased more in the presence of etofibrate than in the presence of equimolecular doses of nicotinic acid or clofibrate. The first drug was the only one to stimulate the rate of fatty acid re-esterification when incubations were done under basal conditions. By 3 h after their acute oral administration all three drugs decreased plasma FFA levels, although the effect from etofibrate was largest, the drugs enhanced or decreased plasma glycerol levels depending on both the dose and the time after treatment. Plasma triglycerides also decreased at 3 h after oral drug administration, and this effect was similar with etofibrate and nicotinic acid but less with clofibrate. With the exception of a decrease at 7 h after the highest dose (1.2 mmol/kg) of either etofibrate or nicotinic acid (but not clofibrate), plasma cholesterol levels remained stable at 7 h after the respective treatments. Thus, the hypocholesterolemic effect of these drugs seems secondary to their hypotriglyceridemic effect, which would be a consequence of their respective antilipolytic actions, and follows an efficiency sequence of etofibrate, nicotinic acid and clofibrate.

13.
J Steroid Biochem Mol Biol ; 53(1-6): 467-73, 1995 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-7626496

RESUMO

Peroxisome proliferators such as clofibric acid, nafenopin, and WY-14,643 have been shown to activate peroxisome proliferator-activated receptor (PPAR), a member of the steroid nuclear receptor superfamily. We have cloned the cDNA from rat that is homologous to that from mouse, which encodes a 97% similar protein. To search for physiologically occurring activators, we established a transcriptional transactivation assay by stably expressing in CHO cells a chimera of rat PPAR and the human glucocorticoid receptor that activates expression of the placental alkaline phosphatase reporter gene under the control of the mouse mammary tumor virus promoter. 150 microM concentrations of arachidonic or linoleic acid but not of dehydroepiandrosterone, cholesterol, or 25-hydroxy-cholesterol, activated the receptor chimera. In addition, saturated fatty acids induced the reporter gene. Shortening the chain length to n = 6 or introduction of an omega-terminal carboxylic group abolished the activation potential of the fatty acid. To test whether a common PPAR binding metabolite might be formed from free fatty acids we tested the effects of differentially beta-oxidizable fatty acids and inhibitors of fatty acid metabolism. The peroxisomal proliferation-inducing, non-beta-oxidizable, tetradecylthioacetic acid activated PPAR to the same extent as the strong peroxisomal proliferator WY-14,643, whereas the homologous beta-oxidizable tetradecylthiopropionic acid was only as potent as a non-substituted fatty acid. Cyclooxygenase inhibitors, radical scavengers or cytochrome P450 inhibitors did not affect activation of PPAR. In conclusion, beta-oxidation is apparently not required for the formation of the PPAR-activating molecule and this moiety might be a fatty acid, its ester with CoA, or a further derivative of the activated fatty acid prior to beta-oxidation of the acyl-CoA ester.


Assuntos
Ácidos Graxos/metabolismo , Receptores Citoplasmáticos e Nucleares/metabolismo , Fatores de Transcrição/metabolismo , Animais , Sistema Enzimático do Citocromo P-450/genética , Regulação da Expressão Gênica , Ligantes , Camundongos , Microcorpos/fisiologia , Regiões Promotoras Genéticas , Ratos , Transdução de Sinais
14.
Gen Pharmacol ; 26(3): 537-42, 1995 May.
Artigo em Inglês | MEDLINE | ID: mdl-7789727

RESUMO

1. The effect of etofibrate, the ethandiol-1,2 diester of nicotinic and clofibric acids on bile production was studied in male rats that received a daily dose of 300 mg of etofibrate/kg body weight by stomach tube for 10 days and were compared with control rats receiving the medium. 2. The bile duct was cannulated, animals were intravenously given 1 microCi (4-14C)-cholesterol/100 b.w. and bile was collected at different intervals for a total of 4 hr. 3. Etofibrate treatment decreased plasma cholesterol and triglyceride concentrations and increased the bile flow. The cummulative amount of both bile volume and total bile radioactivity secreted increased linearly in all the animals; the respective slopes being higher in etofibrate treated rats than in controls. 4. The main labelled component found in the bile was always bile acids rather than cholesterol and the proportion of each of these compounds was similar in both groups. Neither was any difference between the groups found in the concentration of bile acids, cholesterol and phospholipids nor in the cholesterol/(bile+phospholipid) ratio. 5. Besides other factors, the present results indicate that an increase in bile flow and biliary cholesterol excretion in its free form and after its conversion into bile acids should contribute to the hypocholesterolemic effect of etofibrate.


Assuntos
Anticolesterolemiantes/farmacologia , Bile/metabolismo , Ácido Clofíbrico/análogos & derivados , Lipídeos/fisiologia , Animais , Bile/efeitos dos fármacos , Peso Corporal/efeitos dos fármacos , Colesterol/sangue , Colesterol/metabolismo , Ácido Clofíbrico/farmacologia , Lipídeos/sangue , Masculino , Tamanho do Órgão/efeitos dos fármacos , Fosfolipídeos/metabolismo , Ratos , Ratos Sprague-Dawley , Triglicerídeos/sangue
15.
Biochim Biophys Acta ; 1168(3): 340-7, 1993 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-8323974

RESUMO

To contribute to the understanding of the hypolipidemic action of etofibrate, which is the 1,2-ethandiol ester of clofibric acid and nicotinic acid, 300 mg of this drug/kg body weight or of the medium were administered daily by a stomach tube to normolipidemic rats. Some animals were decapitated at the 10th day of daily treatment (prolonged treatment), whereas others were studied at different times after one single administration (acute treatment). In animals on prolonged treatment etofibrate decreased plasma levels of cholesterol, triacylglycerols, free fatty acids (FFA) and glycerol, as well as the total and unesterified cholesterol concentrations, in liver microsomes. In these rats, etofibrate increased the activity of liver cytosolic glycerol-3-P dehydrogenase, whereas it decreased the activity of both microsomal HMG-CoA reductase and cholesterol 7 alpha-hydroxylase and did not affect acyl-CoA: cholesterol acyltransferase (ACAT). At 3, 5 and 7 h after acute treatment, etofibrate decreased plasma levels of triacylglycerols, glycerol and FFA, and this effect disappeared at 24 h, whereas plasma cholesterol did not change 3 h after etofibrate but decreased at 5 and 7 h and remained low after 24 h, and a similar change was found in the liver microsomes free cholesterol concentration. However, with the exception of a significant reduction in cytosolic glycerol-3-P dehydrogenase at 7 h and in ACAT at 5 h, acute etofibrate treatment did not affect the activity of the liver enzymes studied. At low concentrations (10(-5) M) in the incubation medium, etofibrate decreased the release of both FFA and glycerol by epididymal fat pad pieces incubated in vitro. These findings together with those previously reported by us in rats using a similar etofibrate treatment protocol [6] indicate that etofibrate decreases the availability of lipolytic products in the liver by acting on their release from adipose tissue and on their intrinsic hepatic metabolism. Consequently, this drug would decrease liver VLDL triacylglycerol synthesis and secretion, which together with facilitating the clearance of circulating triacylglycerols causes its hypotriglyceridemic effect. The hypocholesterolemic effect of etofibrate after acute treatment may be a secondary consequence of the reduced liver VLDL production caused by decreased adipose tissue lipolysis, but after prolonged treatment, this effect also seems to be influenced by the inhibition of HMG-CoA reductase activity which would reduce cholesterol synthesis.


Assuntos
Tecido Adiposo/metabolismo , Ácido Clofíbrico/análogos & derivados , Hipolipemiantes/farmacologia , Lipídeos/sangue , Fígado/enzimologia , Animais , Peso Corporal , Ácido Clofíbrico/administração & dosagem , Ácido Clofíbrico/farmacologia , Hipolipemiantes/administração & dosagem , Lipólise/efeitos dos fármacos , Masculino , Tamanho do Órgão , Ratos , Ratos Sprague-Dawley , Frações Subcelulares/metabolismo , Fatores de Tempo
16.
Rev Esp Fisiol ; 48(3): 211-4, 1992 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-1301638

RESUMO

A relatively little time consuming simple method based on the treatment of bile with cholesterol oxidase and subsequent high performance liquid chromatography measurement of the 3-ketocholesterol produced in order to determine the level of the cholesterol concentration is described. The method avoids bilirubin interferences, has high reproducibility and recovery assays give 100% values. It is highly sensitive and suitable for use in the determination of cholesterol concentrations in bile and other bilirubin containing biological fluids.


Assuntos
Bile/química , Colesterol/análise , Cromatografia Líquida de Alta Pressão , Cetocolesteróis/análise , Animais , Masculino , Ratos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Fatores de Tempo
17.
J Lipid Res ; 33(6): 931-6, 1992 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-1512516

RESUMO

The study of the cellular events that lead to the foam cell formation requires the development of fast, accurate, and sensitive methods to quantify cholesterol in cultured cells. Here we describe a procedure that allows the rapid determination of free and total cholesterol in a reduced number of cells, which makes it very suitable for cholesterol determination in cell cultures. The method consists of the enzymatic conversion of cholesterol to cholest-4-ene-3-one by cholesterol oxidase followed by the analysis of the sample by high performance liquid chromatography (HPLC) to detect this oxidized product. Due to the relatively high wavelength at which cholest-4-ene-3-one has its maximum absorption (240 nm), other cellular components do not interfere with the chromatographic procedure and prior lipid extraction is not required. Moreover, the duration of each chromatogram is about 3 min, contributing to the celerity of the method. All the cholesteryl esters used (oleate, palmitate, stearate and linoleate) were quantitatively hydrolyzed by incubation with cholesterol esterase; this was observed to occur with both pure standards and in cell homogenates. Sensitivity is enough to allow the determination of free and total cholesterol in less than 5 x 10(3) cells. We have applied this method to human monocyte-derived macrophages and the values obtained for free and total cholesterol are in close agreement with published data.


Assuntos
Colesterol Oxidase/análise , Colesterol/análise , Cromatografia Líquida de Alta Pressão/métodos , Monócitos/química , Células Cultivadas , Colestenonas/análise , Colesterol/metabolismo , Ésteres do Colesterol/metabolismo , Humanos , Hidrólise , Indicadores e Reagentes
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