RESUMO
Bile acids act as signalling molecules that contribute to maintenance of energy homeostasis in mice and humans. Activation of G-protein-coupled bile acid receptor TGR5 induces energy expenditure in brown adipose tissue (BAT). However, a role for the nuclear bile acid receptor Farnesoid X receptor (FXR) in BAT has remained ambiguous. We aimed to study the potential role of FXR in BAT development and functioning. Here we demonstrate low yet detectable expression of the α1/2 isoforms of FXR in murine BAT that markedly decreases upon cold exposure. Moderate adipose tissue-specific FXR overexpression in mice induces pronounced BAT whitening, presenting with large intracellular lipid droplets and extracellular collagen deposition. Expression of thermogenic marker genes including the target of Tgr5, Dio2, was significantly lower in BAT of chow-fed aP2-hFXR mice compared to wild-type controls. Transcriptomic analysis revealed marked up-regulation of extracellular matrix formation and down-regulation of mitochondrial functions in BAT from aP2-hFXR mice. In addition, markers of cell type lineages deriving from the dermomyotome, such as myocytes, as well as markers of cellular senescence were strongly induced. The response to cold and ß3-adrenergic receptor agonism was blunted in these mice, yet resolved BAT whitening. Newborn cholestatic Cyp2c70-/- mice with a human-like bile acid profile also showed distinct BAT whitening and upregulation of myocyte-specific genes, while thermogenic markers were down-regulated. Ucp1 expression inversely correlated with plasma bile acid levels. Therefore, bile acid signalling via FXR has a role in BAT function already early in tissue development. Functionally, FXR activation appears to oppose TGR5-mediated thermogenesis.
Assuntos
Tecido Adiposo Marrom , Receptores Acoplados a Proteínas G , Camundongos , Humanos , Animais , Recém-Nascido , Tecido Adiposo Marrom/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Ácidos e Sais Biliares/metabolismo , Transdução de SinaisRESUMO
INTRODUCTION: The importance of gut microbiome in cardiovascular disease has been increasingly recognized. Trimethylamine N-oxide (TMAO) is a gut microbe-derived metabolite that is associated with cardiovascular disease, including atrial fibrillation (AF). The role of TMAO in clinical AF progression however remains unknown. METHODS AND RESULTS: In this study we measured TMAO and its precursor (betaine, choline, and L- carnitine) levels in 78 patients using plasma samples from patients that participated in the AF-RISK study. 56 patients suffered from paroxysmal AF and 22 had a short history of persistent AF. TMAO levels were significantly higher in patients with persistent AF, as compared to those with paroxysmal AF (median [IQR] 5.65 [4.7-9.6] m/z versus 4.31 [3.2-6.2] m/z, p < 0.05), while precursor levels did not differ. In univariate analysis, we observed that for every unit increase in TMAO, the odds for having persistent AF increased with 0.44 [0.14-0.73], p < 0.01. Conclusion: These results suggest that higher levels of TMAO are associated with more progressed forms of AF. We therefore hypothesize that increased TMAO levels may reflect disease progression in humans. Larger studies are required to validate these preliminary findings.Trial Registration number: Clinicaltrials.gov NCT01510210.
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BACKGROUND AND OBJECTIVES: The antiobesity effects of suppressed endocannabinoid signaling may rely, at least in part, on changes in lipid fluxes. As fatty acids exert specific effects depending on their level of saturation, we hypothesized that the dietary fatty acid composition would influence the outcome of treatment with a CB(1)-receptor antagonist (rimonabant). METHODS: Mice were treated with rimonabant (10 mg kg(-1) body weight per day) or vehicle while equicalorically fed either a low-fat diet (LF), a high-fat (HF) diet or an HF diet in which 10% of the saturated fatty acids (SFAs) were replaced by poly-unsaturated fatty acids (PUFA) from fish oil (FO). Food intake and body weight were registered daily. Indirect calorimetry was performed and feces were collected. After 3 weeks, mice were killed for blood and tissue collection. RESULTS: Relative to the LF diet, the HF diet caused anticipated metabolic derangements, which were partly reversed by the HF/FO diet. The HF/FO diet, however, was most obesity-promoting despite inhibiting lipogenesis as indicated by low gene expression levels of lipogenic enzymes. On all three diets, rimonabant treatment improved metabolic derangements and led to significantly lower body weight gain than their respective controls. This latter effect appeared largest in the HF/FO group, but occurred without major changes in nutrient absorption and energy expenditure. CONCLUSION: The effects of chronic rimonabant treatment on body weight gain occurred irrespective of diet-induced changes in lipogenic activity, food intake and daily energy expenditure, and were, in fact, most pronounced in HF/FO mice. The effects of dietary PUFA replacement in an HF diet on expansion of adipose tissue might allow the favorable effects of dietary PUFA on dyslipidemia and hepatic steatosis. In light of other disadvantageous effects of weight gain, this might be a risky trade-off.
Assuntos
Peso Corporal/efeitos dos fármacos , Gorduras na Dieta/metabolismo , Ácidos Graxos/metabolismo , Óleos de Peixe/metabolismo , Obesidade/metabolismo , Piperidinas/administração & dosagem , Pirazóis/administração & dosagem , Animais , Canabinoides/antagonistas & inibidores , Gorduras na Dieta/farmacologia , Metabolismo Energético/efeitos dos fármacos , Óleos de Peixe/administração & dosagem , Regulação Enzimológica da Expressão Gênica , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Obesidade/tratamento farmacológico , Obesidade/genética , Receptor CB1 de Canabinoide/antagonistas & inibidores , RimonabantoRESUMO
AIM: Inhibition of the acetyl-CoA carboxylase (ACC) system, consisting of the isozymes ACC1 and ACC2, may be beneficial for treatment of insulin resistance and/or obesity by interfering with de novo lipogenesis and beta-oxidation. We have evaluated effects of pharmacological inhibition of ACC by soraphen (SP) on high fat (HF) diet-induced insulin resistance in mice. METHOD: Male C57Bl6/J mice were fed control chow, a HF diet or a HF diet supplemented with SP (50 or 100 mg/kg/day). RESULTS: Body weight gain and total body fat content of SP-treated animals were significantly reduced compared with HF-fed mice. Fractional synthesis of palmitate was significantly reduced in mice treated with SP, indicative for ACC1 inhibition. Plasma beta-hydroxybutyrate levels were significantly elevated by SP, reflecting simultaneous inhibition of ACC2 activity. Mice treated with SP showed improved peripheral insulin sensitivity, as assessed by hyperinsulinaemic euglycaemic clamps. CONCLUSION: Pharmacological inhibition of the ACC system is of potential use for treatment of key components of the metabolic syndrome.
Assuntos
Acetil-CoA Carboxilase/antagonistas & inibidores , Dieta , Gorduras na Dieta/administração & dosagem , Resistência à Insulina/fisiologia , Macrolídeos/farmacologia , Ácido 3-Hidroxibutírico/sangue , Animais , Colesterol/metabolismo , Técnica Clamp de Glucose , Insulina/metabolismo , Fígado/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Obesidade/metabolismo , Obesidade/fisiopatologia , Ácido Palmítico/metabolismo , Reação em Cadeia da Polimerase/métodos , RNA/metabolismo , RNA Mitocondrial , Aumento de Peso/efeitos dos fármacosRESUMO
Glycogen storage disease type Ia (GSD Ia) is characterized by severe hypercholesterolaemia and hypertriglyceridaemia. Little is known about the aetiology of the hyperlipidaemia in GSD Ia. Adipokines play an important regulatory role in lipid metabolism. We investigated whether adipokine concentrations were correlated with the degree of hyperlipidaemia in GSD Ia patients. Six patients with GSD Ia were studied in semi-fasted conditions. Adiponectin, but not leptin, correlated (r(2) = -0.79, p = 0.02) with plasma triglyceride concentrations in the GSD Ia patients. Leptin correlated well with BMI (r(2) = 0.59, p < 0.01). However, neither body mass index (BMI) nor homeostasis model assessment (HOMA), as a marker of insulin sensitivity, correlated with triglyceride concentrations. Although a small number of patients were studied, these results indicate that adiponectin concentrations are correlated with the degree of hypertriglyceridaemia in GSD Ia. Pharmacological treatment aimed at increasing adiponectin levels might improve the metabolic status of these patients.
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Adiponectina/sangue , Doença de Depósito de Glicogênio Tipo I/sangue , Hipertrigliceridemia/sangue , Adolescente , Adulto , Índice de Massa Corporal , Estudos de Casos e Controles , Feminino , Doença de Depósito de Glicogênio Tipo I/complicações , Doença de Depósito de Glicogênio Tipo I/patologia , Humanos , Hipertrigliceridemia/tratamento farmacológico , Hipertrigliceridemia/etiologia , Resistência à Insulina , Leptina/sangue , Masculino , Índice de Gravidade de Doença , Tiazolidinedionas/farmacologia , Triglicerídeos/sangue , Adulto JovemRESUMO
Transport systems involved in uptake and biliary secretion of bile salts have been extensively studied in rat liver; however, little is known about these systems in the human liver. In this study, we investigated taurocholate (TC) transport in canalicular and basolateral plasma membrane vesicles isolated from 15 human livers (donor age 6-64 yr). ATP stimulated the uptake of TC into both canalicular and basolateral human liver plasma membrane vesicles (cLPM and blLPM, respectively). Considerable interindividual variations in the transport velocity were observed in the different membrane preparations used: 9.0 +/- 1.3 (mean +/- SEM, n = 17; range 1.6-18.0) and 9.3 +/- 2.0 (range 1.1-29.8) pmol TC.mg protein-1.min-1 at 1.0 microM TC for cLPM and blLPM, respectively. TC transport was temperature sensitive and showed saturation kinetics with a high affinity for TC (Km 4.2 +/- 0.7 microM and 3.7 +/- 0.5 microM for cLPM and blLPM, respectively). Transport was dependent on the ATP concentration and saturable (Km 0.25 +/- 0.03 mM, n = 3). Neither nitrate, which reduces membrane potential, nor the protonophore FCCP strongly inhibited ATP-dependent TC transport, indicating that membrane potential and proton gradient are not involved in this process. TC transport was significantly inhibited by the classical anion transport inhibitor 4,4'-diisothiocyanostilbene-2,2'-disulfonate (250 microM) and the glutathione conjugate S-(2,4-dinitrophenyl)glutathione (100 microM). In conclusion, high affinity ATP-dependent TC transport is present in human liver at both the canalicular and the basolateral sides of the hepatocyte.
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Trifosfato de Adenosina/metabolismo , Fígado/metabolismo , Ácido Taurocólico/metabolismo , Adulto , Fatores Etários , Transporte Biológico Ativo/efeitos dos fármacos , Membrana Celular/metabolismo , Criança , Pré-Escolar , Feminino , Humanos , Técnicas In Vitro , Cinética , Masculino , Pessoa de Meia-IdadeRESUMO
Biliary secretion of 3 alpha-sulfated bile acids has been studied in Wistar rats with an autosomal recessive defect in the hepatic transport of bilirubin. Liver function, established by measurement of various enzymes in plasma, by enzyme histochemical methods, and by electron microscopy, appeared to be normal in these rats. Serum levels of unconjugated, monoglucuronidated, and diglucuronidated bilirubin were 0.62, 1.62, and 6.16 mumol/liter, respectively, compared with 0.17, 0.08, and 0.02 mumol/liter in control rats. Biliary bilirubin secretion was strongly reduced in the mutant animals: 0.21 +/- 0.03 vs. 0.39 +/- 0.03 nmol/min per 100 g body wt in control rats. Despite normal biliary bile acid output, bile flow was markedly impaired in the mutant animals, due to a 53% reduction of the bile acid-independent fraction of bile flow. The transport maximum for biliary secretion of dibromosulphthalein (DBSP) was also drastically reduced (-53%). Biliary secretion of intravenously administered trace amounts of the 3 alpha-sulfate esters of 14C-labeled taurocholic acid (-14%), taurochenodeoxycholic acid (-39%), taurolithocholic acid (-73%), and glycolithocholic acid (-91%) was impaired in the jaundiced rats compared with controls, in contrast to the biliary secretion of the unsulfated parent compounds. Hepatic uptake of sulfated glycolithocholic acid was not affected in the jaundiced animals. Preadministration of DBSP (15 mumol/100 g body wt) to normal Wistar rats significantly impaired the biliary secretion of sulfated glycolithocholic acid, but did not affect taurocholic acid secretion. We conclude that separate transport systems in the rat liver exist for biliary secretion of sulfated and unsulfated bile acids; the sulfates probably share secretory pathways with the organic anions bilirubin and DBSP. The described genetic defect in hepatic transport function is associated with a reduced capacity to secrete sulfated bile acids into bile; this becomes more pronounced with a decreasing number of hydroxyl groups on the sulfated bile acid's molecule.
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Ácidos e Sais Biliares/metabolismo , Bilirrubina/metabolismo , Fígado/metabolismo , Animais , Bile/fisiologia , Bilirrubina/sangue , Transporte Biológico , Histocitoquímica , Ácido Litocólico/análogos & derivados , Ácido Litocólico/metabolismo , Fígado/ultraestrutura , Testes de Função Hepática , Masculino , Microscopia Eletrônica , Ratos , Ratos Endogâmicos , Sulfobromoftaleína/análogos & derivados , Sulfobromoftaleína/metabolismo , Ácido Tauroquenodesoxicólico/metabolismo , Ácido Taurocólico/metabolismo , Ácido Taurolitocólico/metabolismoRESUMO
In cultured hepatocytes conversion of [4-14C]cholesterol into bile acids was dose dependently reduced by the antimycotic drug ketoconazole, giving half-maximal inhibition at 10 microM ketoconazole in rat hepatocytes and at 1 microM in human hepatocytes. No change was observed in the ratio of produced cholic, beta-muricholic, and chenodeoxycholic acid with increasing amounts of the drug. Conversion of [4-14C]7 alpha-hydroxycholesterol, an intermediate of bile acid pathway, to bile acids was not affected by ketoconazole. These results together with kinetic studies with rat liver microsomes, demonstrating noncompetitive inhibition (Ki = 0.4 microM), indicate that cholesterol 7 alpha-hydroxylase is the main site of inhibition. In bile-diverted rats a single dose of ketoconazole (50 mg/kg) dramatically impaired bile flow and biliary bile acid output (92% inhibition). A similar blockade was observed using [4-14C]cholesterol as precursor for bile acid synthesis. Therefore, treatment of patients with this drug may inhibit bile acid synthesis, resulting in a reduction of the bile acid pool size after long-term ketoconazole therapy.
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Ácidos e Sais Biliares/biossíntese , Colesterol 7-alfa-Hidroxilase/antagonistas & inibidores , Cetoconazol/farmacologia , Fígado/enzimologia , Esteroide Hidroxilases/antagonistas & inibidores , Animais , Bile/metabolismo , Células Cultivadas , Colesterol/metabolismo , Fígado/citologia , Masculino , Microssomos Hepáticos/enzimologia , Ratos , Ácido Taurocólico/metabolismoRESUMO
The ABC transporter ABCA1 regulates HDL levels and is considered to control the first step of reverse cholesterol transport from the periphery to the liver. To test this concept, we studied the effect of ABCA1 deficiency on hepatic metabolism and hepatobiliary flux of cholesterol in mice. Hepatic lipid contents and biliary secretion rates were determined in Abca1(-/-), Abca1(+/-), and Abca1(+/+) mice with a DBA background that were fed either standard chow or a high-fat, high-cholesterol diet. Hepatic cholesterol and phospholipid contents in Abca1(-/-) mice were indistinguishable from those in Abca1(+/-) and Abca1(+/+) mice on both diets. In spite of the absence of HDL, biliary secretion rates of cholesterol, bile salts, and phospholipid were unimpaired in Abca1(-/-) mice. Neither the hepatic expression levels of genes controlling key steps in cholesterol metabolism nor the contribution of de novo synthesis to biliary cholesterol and bile salts were affected by Abca genotype. Finally, fecal excretion of neutral and acidic sterols was similar in all groups. We conclude that plasma HDL levels and ABCA1 activity do not control net cholesterol transport from the periphery via the liver into the bile, indicating that the importance of HDL in reverse cholesterol transport requires re-evaluation.
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Transportadores de Cassetes de Ligação de ATP/fisiologia , Sistema Biliar/metabolismo , Proteínas de Transporte , Colesterol/metabolismo , Lipoproteínas HDL/deficiência , Fígado/metabolismo , Proteínas de Membrana , Proteínas de Ligação a RNA , Receptores Imunológicos , Transportador 1 de Cassete de Ligação de ATP , Transportadores de Cassetes de Ligação de ATP/genética , Animais , Transporte Biológico Ativo , Antígenos CD36/genética , Humanos , Hidroximetilglutaril-CoA Redutases/genética , Lipoproteínas HDL/metabolismo , Camundongos , Camundongos Endogâmicos DBA , Camundongos Knockout , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Receptores de LDL/genética , Receptores de Lipoproteínas/genética , Receptores Depuradores , Receptores Depuradores Classe BRESUMO
Hepatic cholesterol metabolism was studied in rats fed purified diets supplemented (9% wt/wt) with either fish oil (FO) (n-3 fatty acids) or corn oil (CO) (n-6 fatty acids) for 4 wk. Rats were equipped with permanent catheters in heart, bile duct, and duodenum to allow studies under normal feeding conditions. [3H]-cholesteryl oleate-labeled small unilamellar liposomes, which are rapidly endocytosed by hepatocytes, were intravenously injected to label intrahepatic cholesterol pools, and plasma and bile were collected. FO as compared to CO induced a lowering of plasma cholesterol levels by 38% and of triglyceride levels by 69%. This reduction in plasma lipids in FO rats was accompanied by: (a) an increased bile acid pool size (28%); (b) a fourfold increase in the ratio cholic acid/chenodeoxycholic acid in bile; (c) increased biliary excretion of cholesterol (51%); (d) accelerated excretion of endocytosed free cholesterol into bile; (e) accelerated incorporation of endocytosed cholesterol in bile acids; (f) a significant increase in the bile acid-independent fraction of bile flow; and (g) a threefold increase in hepatic alkaline phosphatase activity. The results show that FO induces changes in transport and metabolic pathways of cholesterol in the rat liver, which result in a more rapid disposition of plasma-derived cholesterol into the bile.
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Ácidos e Sais Biliares/biossíntese , Colesterol/metabolismo , Gorduras na Dieta/farmacologia , Óleos de Peixe/farmacologia , Fígado/metabolismo , Fosfatase Alcalina/análise , Animais , Ácidos e Sais Biliares/análise , Transporte Biológico , Óleo de Milho/farmacologia , Lipídeos/sangue , Lipoproteínas/metabolismo , Masculino , Ratos , Ratos EndogâmicosRESUMO
The process of hepatobiliary copper (Cu) secretion is still poorly understood: Cu secretion as a complex with glutathione and transport via a lysosomal pathway have been proposed. The recent cloning and sequencing of the gene for Wilson disease indicates that Cu transport in liver cells may be mediated by a Cu transporting P-type ATPase. Biochemical evidence for ATP-dependent Cu transport in mammalian systems, however, has not been reported so far. We have investigated Cu transport in rat liver plasma membrane vesicles enriched in canalicular or basolateral membranes in the presence and absence of ATP (4 mM) and an ATP-regenerating system. The presence of ATP clearly stimulated uptake of radiolabeled Cu (64Cu, 10 microM) into canalicular plasma membrane vesicles and, to a lesser extent, also into basolateral plasma membrane vesicles. ATP-dependent Cu transport was dose-dependently inhibited by the P-type ATPase inhibitor vanadate, and showed saturation kinetics with an estimated Km of 8.6 microM and a Vmax of 6.9 nmol/min/mg protein. ATP-stimulated Cu uptake was similar in canalicular membrane vesicles of normal Wistar rats and those of mutant GY rats, expressing a congenital defect in the activity of the ATP-dependent canalicular glutathione-conjugate transporter (cMOAT). These studies demonstrate the presence of an ATP-dependent Cu transporting system in isolated plasma membrane fractions of rat liver distinct from cMOAT.
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Trifosfato de Adenosina/metabolismo , Membrana Celular/metabolismo , Cobre/metabolismo , Fígado/metabolismo , Transportadores de Cassetes de Ligação de ATP/genética , Transportadores de Cassetes de Ligação de ATP/metabolismo , Adenosina Trifosfatases/metabolismo , Animais , Transporte Biológico , Fracionamento Celular , Degeneração Hepatolenticular , Masculino , Síndrome dos Cabelos Torcidos , Ratos , Ratos Mutantes , Ratos Wistar , Ácido Taurocólico/metabolismoRESUMO
To explore mechanisms underlying triglyceride (TG) accumulation in livers of chow-fed apo E-deficient mice (Kuipers, F., J.M. van Ree, M.H. Hofker, H. Wolters, G. In't Veld, R.J. Vonk, H.M.G. Princen, and L.M. Havekes. 1996. Hepatology. 24:241-247), we investigated the effects of apo E deficiency on secretion of VLDL-associated TG (a) in vivo in mice, (b) in isolated perfused mouse livers, and (c) in cultured mouse hepatocytes. (a) Hepatic VLDL-TG production rate in vivo, determined after Triton WR1339 injection, was reduced by 46% in apo E-deficient mice compared with controls. To eliminate the possibility that impaired VLDL secretion is caused by aspecific changes in hepatic function due to hypercholesterolemia, VLDL-TG production rates were also measured in apo E-deficient mice after transplantation of wild-type mouse bone marrow. Bone marrow- transplanted apo E-deficient mice, which do not express apo E in hepatocytes, showed normalized plasma cholesterol levels, but VLDL-TG production was reduced by 59%. (b) VLDL-TG production by isolated perfused livers from apo E-deficient mice was 50% lower than production by livers from control mice. Lipid composition of nascent VLDL particles isolated from the perfusate was similar for both groups. (c) Mass VLDL-TG secretion by cultured apo E-deficient hepatocytes was reduced by 23% compared with control values in serum-free medium, and by 61% in the presence of oleate in medium (0. 75 mM) to stimulate lipogenesis. Electron microscopic evaluation revealed a smaller average size for VLDL particles produced by apo E-deficient cells compared with control cells in the presence of oleate (38 and 49 nm, respectively). In short-term labeling studies, apo E-deficient and control cells showed a similar time-dependent accumulation of [3H]TG formed from [3H]glycerol, yet secretion of newly synthesized VLDL-associated [3H]TG by apo E-deficient cells was reduced by 60 and 73% in the absence and presence of oleate, respectively. We conclude that apo E, in addition to its role in lipoprotein clearance, has a physiological function in the VLDL assembly-secretion cascade.
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Apolipoproteínas E/deficiência , Lipoproteínas VLDL/metabolismo , Fígado/metabolismo , Triglicerídeos/metabolismo , Animais , Apolipoproteínas E/fisiologia , Transplante de Medula Óssea , Células Cultivadas , Feminino , Técnicas In Vitro , Lipoproteínas VLDL/biossíntese , Fígado/citologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Perfusão , Triglicerídeos/biossínteseRESUMO
Cholesterol is an important sterol in mammals. Defects in cholesterol synthesis or intracellular routing have devastating consequences already in utero: the Smith-Lemli-Opitz syndrome, desmosterolosis and Niemann-Pick C1 disease provide examples of severe human inherited diseases caused by mutations in cholesterol metabolism genes. On the other hand, elevated plasma cholesterol concentrations are associated with the development of atherosclerosis which represents a major health risk in Western societies. Moreover, several studies indicate that development of atherosclerosis may already start during fetal life. Hence, a carefully balanced regulation of cholesterol metabolism appears of critical importance for both the development of the fetus and health of the adult. In the adult, the liver X receptor is a key regulator of cholesterol metabolism. Its target genes regulate cellular cholesterol efflux and thereby modulate whole-body cholesterol fluxes. LXR and several of its target genes have recently been demonstrated to be expressed in the placenta, which would provide a means to control delivery of maternal cholesterol to the fetus. Here we discuss the potential role of the placenta in the regulation of fetal cholesterol homeostasis and strategies to influence maternal-fetal cholesterol transfer.
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Colesterol/metabolismo , Proteínas de Ligação a DNA/metabolismo , Feto/metabolismo , Troca Materno-Fetal , Placenta/metabolismo , Receptores Citoplasmáticos e Nucleares/metabolismo , Animais , Transporte Biológico , Proteínas de Ligação a DNA/efeitos dos fármacos , Proteínas de Ligação a DNA/genética , Feminino , Humanos , Receptores X do Fígado , Troca Materno-Fetal/efeitos dos fármacos , Doenças de Niemann-Pick/genética , Receptores Nucleares Órfãos , Gravidez , Receptores Citoplasmáticos e Nucleares/efeitos dos fármacos , Receptores Citoplasmáticos e Nucleares/genética , Síndrome de Smith-Lemli-Opitz/genéticaRESUMO
BACKGROUND: A high-fat diet has been recognized for some time as a major risk factor for colorectal cancer. It is thought that fat promotes this disease by increasing the levels of fatty and bile acids within the colon. These acids irritate and damage the epithelial cells of the colon. As a result of this cellular destruction, an increase in the rate of cellular proliferation occurs. Oral calcium supplementation has been proposed as a dietary intervention for individuals at high risk of colorectal cancer because of its ability to reduce rectal epithelial cell proliferation through the binding of fatty and bile acids. Placebo-controlled studies, however, have yielded varying results. PURPOSE: We conducted a randomized, double-blinded, placebo-controlled trial to test oral calcium supplementation in patients at high risk of developing hereditary nonpolyposis colorectal cancer. METHODS: Thirty subjects at risk for this cancer, with an increased epithelial cell proliferation along the colon and rectum, were randomly assigned to either a placebo group (n = 15) or a treatment group (n = 15). They received either oral calcium carbonate (CaCO3) supplements (1.5 g) or placebo (cellulose and starch) three times a day during a 12-week period. Colonic biopsy specimens (rectal, sigmoidal, and descending) were obtained prior to and after the intervention trial, during endoscopy, for determination of labeling index (LI) of whole crypts and crypt compartments by 5-bromo-2'-deoxyuridine incorporation and immunohistochemistry. Proportional bile acid compositions in duodenal bile and cytolytic activity of fecal water were also determined. All P values represent two-tailed tests of statistical significance. RESULTS: Statistically significant reductions, comparing before with after intervention, in rectal whole-crypt LI after receiving either calcium supplements (from 10.9% +/- 5.2% [mean +/- SD] to 6.2% +/- 1.5%; P < .02) or placebo (from 11.7% +/- 4.7% to 8.2% +/- 3.1%; P < .05) were observed. In the three bowel segments, no statistically significant differences were observed between the supplemental calcium and placebo groups. A statistically significant reduction in cytolytic activity was determined during calcium supplementation (from 57% +/- 41% to 32% +/- 30%; P < .05), whereas in the placebo group, it did not change (from 42% +/- 41% to 36% +/- 27%; P > .10). CONCLUSIONS: Oral calcium supplementation was shown to cause only a minor nonstatistically significant reduction of epithelial cell proliferation in the rectum, compared with placebo, and to have no effect on the same parameter in the sigmoid and descending colon in first-degree relatives of hereditary nonpolyposis colorectal cancer patients. IMPLICATION: These results cast doubt on the value of calcium supplementation in the prevention of colorectal cancer, especially in individuals already consuming an adequate amount of dietary calcium.
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Cálcio/uso terapêutico , Neoplasias Colorretais Hereditárias sem Polipose/tratamento farmacológico , Adolescente , Adulto , Bile/química , Divisão Celular , Neoplasias Colorretais Hereditárias sem Polipose/patologia , Método Duplo-Cego , Células Epiteliais , Fezes/química , Feminino , Humanos , Mucosa Intestinal/citologia , Masculino , Pessoa de Meia-Idade , PlacebosRESUMO
Subtotal colectomy and ileorectal anastomosis in familial adenomatous polyposis patients can induce temporary regression of adenomas in the rectum. The mechanism for this phenomenon is unclear. We evaluated the effect of colectomy on rectal mucosal proliferation, in relation to changes in bile acid metabolism. Four familial adenomatous polyposis patients were studied before and 3-6 months after surgery, and eight others 7-22 years postoperatively. Within 6 months after surgery, the size of the proliferative zone of the colonic crypts was found to be reduced (P less than 0.05). The proliferative activity of total colonic crypts was not affected within this period. More than 7 years postoperatively, increased cell proliferation of total crypts (P less than 0.02), as well as mid (P less than 0.05) and basal (P less than 0.05) crypt compartments, were observed compared to shortly after colectomy. In duodenal bile, deoxycholic acid was absent shortly after operation, whereas several years after operation only a small fraction (2%) was present. Fecal secondary bile acid excretion diminished after colectomy and did not change several years postoperatively. In postoperative stools only, small proportions of ursocholic and ursodeoxycholic acids (about 5% each) were consistently found. As subtotal colectomy causes a temporary decrease in the length of the proliferative zone of rectal crypts toward a normal pattern, this may explain regression of rectal polyps. This temporary effect may be mediated, at least in part, by decreased amounts of cytotoxic secondary bile acids in the rectal lumen.
Assuntos
Polipose Adenomatosa do Colo/cirurgia , Ácidos e Sais Biliares/metabolismo , Colectomia , Mucosa Intestinal/metabolismo , Reto/patologia , Polipose Adenomatosa do Colo/metabolismo , Polipose Adenomatosa do Colo/patologia , Adolescente , Adulto , Divisão Celular , Duodeno/química , Epitélio/patologia , Fezes/química , Humanos , Pessoa de Meia-IdadeRESUMO
To gain insight into the possible physiological mechanisms responsible for the increased incidence of colonic neoplasms in patients with acromegaly, a prospective cohort study was carried out in 30 patients with acromegaly, a prospective cohort study was carried out in 30 patients with acromegaly. Seven patients had newly diagnosed acromegaly and 23 were studied during follow-up. Serum growth hormone and insulin-like growth factor-1 (IGF-1) were determined on two separate occasions. During diagnostic endoscopy, mucosal biopsies were obtained for immunohistochemical determination of sigmoidal epithelial cell proliferation, expressed as labeling index (LI). Duodenal and fecal bile acid analyses were performed using gas-liquid chromatography. Results were compared with normal ranges of the laboratory. An increased overall LI was found in 54% of the patients. Increased LI of the luminal, middle, and basal crypt compartments was found in 11, 64, and 28%, respectively. Similarly, comparisons of the mean +/- SEM of the overall LI and the LI of the middle and basal compartments between acromegalic patients and a control group showed overall LI 10.0 +/- 0.8% versus 5.7 +/- 0.6% (P < 0.001), middle LI 12.1 +/- 1.2% versus 5.0 +/- 0.6% (P < 0.001), and basal LI 17.1 +/- 1.3% versus 10.8 +/- 1.3% (P < 0.01). Duodenal and fecal bile acid proportions were within the normal ranges of the laboratory. There was a positive correlation between growth hormone and overall LI (r = 0.55, P < 0.01) by least square regression analysis. There was no correlation between duodenal bile acid composition and hormone levels. The proportion of secondary bile acids in feces correlated with growth hormone (r = 0.55, P < 0.05) as well as with IGF-1 (r = 0.59, P < 0.05). With multiple regression analyses, only a relation between overall LI and IGF-1 (P = 0.007) remained to hold true. Increased epithelial cell proliferation, most probably due to a direct stimulatory effect of especially IGF-1, contributes to the increased risk of colonic neoplasms in acromegaly.
Assuntos
Acromegalia/complicações , Colo/citologia , Neoplasias do Colo/epidemiologia , Acromegalia/epidemiologia , Acromegalia/metabolismo , Adulto , Idoso , Ácidos e Sais Biliares/metabolismo , Divisão Celular/fisiologia , Estudos de Coortes , Neoplasias do Colo/etiologia , Neoplasias do Colo/patologia , Duodeno/metabolismo , Células Epiteliais , Estudos de Avaliação como Assunto , Fezes/química , Feminino , Seguimentos , Hormônio do Crescimento/sangue , Humanos , Incidência , Fator de Crescimento Insulin-Like I/metabolismo , Masculino , Pessoa de Meia-Idade , Estudos ProspectivosRESUMO
The CNS melanocortin (MC) system is implicated as a mediator of the central effects of leptin, and reduced activity of the CNS MC system promotes obesity in both rodents and humans. Because activation of CNS MC receptors has direct effects on autonomic outflow and metabolism, we hypothesized that food intake-independent mechanisms contribute to development of obesity induced by pharmacological blockade of MC receptors in the brain and that changes in hypothalamic neuropeptidergic systems known to regulate weight gain [i.e., corticotropin-releasing hormone (CRH), cocaine-amphetamine-related transcript (CART), proopiomelanocortin (POMC), and neuropeptide Y (NPY)] would trigger this effect. Relative to vehicle-treated controls, third intracerebroventricular (i3vt) administration of the MC receptor antagonist SHU9119 to rats for 11 d doubled food and water intake (toward the end of treatment) and increased body weight ( approximately 14%) and fat content ( approximately 90%), hepatic glycogen content ( approximately 40%), and plasma levels of cholesterol ( approximately 48%), insulin ( approximately 259%), glucagon ( approximately 80%), and leptin ( approximately 490%), whereas spontaneous locomotor activity and body temperature were reduced. Pair-feeding of i3vt SHU9119-treated animals to i3vt vehicle-treated controls normalized plasma levels of insulin, glucagon, and hepatic glycogen content, but only partially reversed the elevations of plasma cholesterol ( approximately 31%) and leptin ( approximately 104%) and body fat content ( approximately 27%). Reductions in body temperature and locomotor activity induced by i3vt SHU9119 were not reversed by pair feeding, but rather were more pronounced. None of the effects found can be explained by peripheral action of the compound. The obesity effects occurred despite a lack in neuropeptide expression responses in the neuroanatomical range selected across the arcuate (i.e., CART, POMC, and NPY) and paraventricular (i.e., CRH) hypothalamus. The results indicate that reduced activity of the CNS MC pathway promotes fat deposition via both food intake-dependent and -independent mechanisms.
Assuntos
Comportamento Animal/fisiologia , Hipotálamo/metabolismo , Obesidade/metabolismo , Receptores da Corticotropina/metabolismo , Transdução de Sinais/fisiologia , Animais , Comportamento Animal/efeitos dos fármacos , Composição Corporal/efeitos dos fármacos , Temperatura Corporal/efeitos dos fármacos , Colesterol/sangue , Ingestão de Líquidos/efeitos dos fármacos , Ingestão de Alimentos/efeitos dos fármacos , Glucagon/sangue , Hipotálamo/efeitos dos fármacos , Injeções Intraventriculares , Insulina/sangue , Leptina/sangue , Masculino , Hormônios Estimuladores de Melanócitos/administração & dosagem , Atividade Motora/efeitos dos fármacos , Neurotransmissores/genética , Neurotransmissores/metabolismo , Pró-Opiomelanocortina/genética , Pró-Opiomelanocortina/metabolismo , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Receptores da Corticotropina/antagonistas & inibidores , Receptores de Melanocortina , Transdução de Sinais/efeitos dos fármacosRESUMO
We studied the hepatic processing and biliary excretion of metabolites of the radiolabeled cytostatic agent 5-fluoro,-2'-deoxy[6-3H]uridine (FUdR) and its lipophilic derivative FUdR-dipalmitate incorporated in liposomes. After intracardial injection in rats, free FUdR was cleared from the circulation within minutes. When FUdR or FUdR-dipalmitate was encapsulated in multilamellar vesicles (MLVs) composed of distearoylphosphatidylcholine/dipalmitoylphosphatidylglycerol (DSPC/DPPG/CHOL, 10:1), as expected, the clearance of 3H label was substantially delayed; incorporation of 50 mol% cholesterol in the liposomal bilayer caused a 2-fold further reduction in elimination rate. Incorporation of FUdR-dipalmitate in small unilamellar vesicles (SUV) of similar composition produced a several-fold further decrease in elimination rate: more than 40% of the injected dose was still circulating after 6 h. The plasma concentration of free FUdR after administration of liposomal FUdR-dipalmitate was below the detection limit (5 x 10(-8) M) at any time. Although only about 9% of the administered radioactivity was excreted into the bile within 48 h after injection of [3H]FUdR, a rapid initial excretion rate was observed (4% of the injected dose in the first 2 h). The bile-associated radioactivity was identified mainly as the catabolite alpha-fluoro-beta-alanine (FBAL), conjugated with the three major bile acid species present in rat bile, i.e., muricholic acid, cholic acid and chenodeoxycholic acid in a ratio of 1:3:1. Liposome incorporation of FUdR or FUdR-dipalmitate did not affect the nature of the excretory products but caused a significant decrease in the initial rate at which label appeared in the bile (< 2% in 6 h).
Assuntos
Ácidos e Sais Biliares/metabolismo , Bile/metabolismo , Floxuridina/metabolismo , Lipossomos/metabolismo , beta-Alanina/análogos & derivados , Animais , Floxuridina/sangue , Floxuridina/urina , Masculino , Ratos , Ratos Wistar , Trítio , beta-Alanina/farmacologiaRESUMO
Glycolithocholic acid and its sulfated derivative are major metabolites of the secondary bile acid lithocholic acid in man. Both compounds are known to induce cholestasis in experimental animals. We compared the effects of these endogenous hepatotoxins on bile production and biliary lipid composition in rats with chronic biliary drainage. The compounds were administered enterally at relatively low rates (5-50% of the rats' endogenous bile acid secretion in these experiments) to simulate enterohepatic circulation. Both compounds were substantially secreted into bile (more than 90% of dose); sulfated glycolithocholic acid unchanged and glycolithocholic acid after hepatic hydroxylation predominantly in the form of glyco-beta-muricholic acid (cf. Kuipers et al. (1986) Am. J. Physiol. 251, G189-G194). Neither glycolithocholic acid nor its sulfated derivative affected the biliary excretion of endogenous bile acids or bile flow in these experiments. In spite of this, phospholipid and cholesterol secretion were significantly reduced by sulfated glycolithocholic acid but were not altered by glycolithocholic acid. Phospholipid and cholesterol secretion rapidly decreased to 25 and 50% of their initial values, respectively, at biliary output rates of sulfated glycolithocholic acid up to 2 mumol/h, and did not further decrease when this output was increased to 6 mumol/h. Small unilamellar liposomes consisting of cholesterol, [Me-14C]choline-labeled phosphatidylcholine, phosphatidylserine and [3H]cholesteryl oleate in a 5:4:1:0.1 molar ratio were employed to label intrahepatic lipid pools. Administration of sulfated glycolithocholic acid slightly reduced bile acid synthesis from [3H]cholesteryl oleate, but significantly reduced the biliary secretion of [14C]phospholipid. Glycolithocholic acid did not affect the hepatic processing of liposomal lipids. It is concluded that sulfated glycolithocholic acid at low doses causes the uncoupling of biliary lipid secretion from that of bile acids, which might represent in initiating event in sulfated glycolithocholic acid hepatotoxicity.
Assuntos
Ácidos e Sais Biliares/metabolismo , Bile/metabolismo , Colesterol/metabolismo , Ácido Glicocólico/análogos & derivados , Ácido Litocólico/análogos & derivados , Fosfolipídeos/metabolismo , Animais , Bile/efeitos dos fármacos , Ácido Glicocólico/farmacologia , Cinética , Lipossomos , Ácido Litocólico/farmacologia , Masculino , Ratos , Ratos EndogâmicosRESUMO
Hepatic cholesterol metabolism was studied in rats with a permanent biliary drainage. Three cholesterol vehicles were used to discriminate between metabolic pathways of cholesterol in the liver. [3H]Cholesterol was administered intravenously associated with rat serum lipoproteins, multilamellar (MLV) or small unilamellar (SUV) liposomes. The liposomes were made from cholesterol, sphingomyelin and phosphatidylserine in a 5:4:1 molar ratio. Initial blood elimination differed markedly for the three vehicles: 15 min after injection the 3H radioactivity content of blood for MLV, SUV and lipoprotein was 3, 50 and 54% of the injected dose, respectively. After about 30 min, MLV-cholesterol label started to reappear in the blood, probably after processing of the vehicle by the Kupffer cells. For all vehicles about 80% of the cholesterol label had been excreted in bile after 120 h, predominantly as bile acids. Initial biliary excretion was highest for lipoproteins (5.7% at 1 h), followed by MLV and SUV (1.3 and 1.2%, respectively). No differences in the radioactivity of excreted bile acids were detectable between the three vehicles at 12 h after injection. However, at 1 h the radioactivity in the muricholic acid fraction was markedly increased, as compared to the other bile acids after injection of SUV-cholesterol, but not after injection of MLV- or lipoprotein-cholesterol. Also, the glycine/taurine conjugation ratio of bile acids was increased for SUV-cholesterol at 1 h as compared to that for the other two vehicles. Since SUV appear to donate their cholesterol to a pool which preferentially supplies cholesterol for muricholic acid synthesis, we conclude that more than one cholesterol pool exists in the hepatocytes from which cholesterol can be recruited for bile acid synthesis. Zonal heterogeneity might be responsible for the observed differences.