RESUMEN
BACKGROUND: Persistent inflammation exacerbates the progression of Duchenne muscular dystrophy (DMD). The hormone, adiponectin (ApN), which is decreased in the metabolic syndrome, exhibits anti-inflammatory properties on skeletal muscle and alleviates the dystrophic phenotype of mdx mice. Here, we investigate whether ApN retains its anti-inflammatory action in myotubes obtained from DMD patients. We unravel the underlying mechanisms by studying the secretome and the early events of ApN. METHODS: Primary cultures of myotubes from DMD and control patients were treated or not by ApN after an inflammatory challenge. Myokines secreted in medium were identified by cytokine antibody-arrays and ELISAs. The early events of ApN signaling were assessed by abrogating selected genes. RESULTS: ApN retained its anti-inflammatory properties in both dystrophic and control myotubes. Profiling of secretory products revealed that ApN downregulated the secretion of two pro-inflammatory factors (TNFα and IL-17A), one soluble receptor (sTNFRII), and one chemokine (CCL28) in DMD myotubes, while upregulating IL-6 that exerts some anti-inflammatory effects. These changes were explained by pretranslational mechanisms. Earlier events of the ApN cascade involved AdipoR1, the main receptor for muscle, and the AMPK-SIRT1-PGC-1α axis leading, besides alteration of the myokine profile, to the upregulation of utrophin A (a dystrophin analog). CONCLUSION: ApN retains its beneficial properties in dystrophic muscles by activating the AdipoR1-AMPK-SIRT1-PGC-1α pathway, thereby inducing a shift in the secretion of downstream myokines toward a less inflammatory profile while upregulating utrophin. ApN, the early events of the cascade and downstream myokines may be therapeutic targets for the management of DMD.
Asunto(s)
Adiponectina/metabolismo , Antiinflamatorios/metabolismo , Músculo Esquelético/metabolismo , Distrofia Muscular de Duchenne/metabolismo , Proteoma/metabolismo , Adolescente , Adulto , Biomarcadores/metabolismo , Niño , Femenino , Regulación de la Expresión Génica , Humanos , Inflamación/metabolismo , Inflamación/patología , Masculino , Fibras Musculares Esqueléticas/metabolismo , Fibras Musculares Esqueléticas/patología , Distrofia Muscular de Duchenne/genética , Distrofia Muscular de Duchenne/patología , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , ARN Interferente Pequeño/metabolismo , Receptores de Adiponectina/metabolismo , Sirtuina 1/metabolismo , Utrofina/genética , Utrofina/metabolismo , Adulto JovenRESUMEN
OBJECTIVES: (1) To investigate whether modulation of the cannabinoid type 1 receptor (CB1R) directly regulates the production of adiponectin (ApN) and other adipokines in omental adipose tissue (OAT) of obese subjects, (2) to establish in which cellular fraction of OAT the effects of CB1R blockade take place and (3) to unravel the underlying mechanisms. SUBJECTS AND METHODS: OAT was obtained from 30 obese subjects (body mass index: 40.6±1.3 kg m(-2)) undergoing abdominal surgery. Primary cultures of explants or of freshly isolated adipocytes or stromal-vascular cells (SVCs) were used. RESULTS: In OAT explants, the CB1R blocker Rimonabant upregulated ApN gene expression. mRNA abundance of omentin that exhibits insulin-sensitizing properties was upregulated as well. Conversely, mRNA levels of two pro-inflammatory cytokines, macrophage inflammatory protein (MIP)-1ß and interleukin (IL)-7 were downregulated. We next examined where these effects took place within OAT. CB1R expression was similar in both cellular fractions. In isolated mature adipocytes, blockade of CB1R reproduced the increase of ApN mRNA and the decrease of IL-7 mRNA, while inducing a rise of ApN secretion into the medium. In isolated SVC, gene expression of omentin, which is restricted to this fraction, was augmented, while that of MIP-1ß was diminished. Finally, we deciphered the mechanisms leading to ApN regulation by the endocannabinoid system (ES). We first established that ApN regulation was actually mediated by the CB1R: ApN gene expression was upregulated by Rimonabant and downregulated by the CB1R agonist arachidonyl-2-chloroethylamide (ACEA). Upregulation of ApN by Rimonabant was unaltered by inhibiting cAMP production. However, downregulation of ApN by ACEA was fully reversed by an inhibitor of p38 mitogen-activated protein kinase (p38MAPK) and ACEA increased p38MAPK phosphorylation. CONCLUSIONS: Blockade of CB1R attenuates the inflammatory state in both cellular fractions of OAT either by increasing ApN and omentin production or by decreasing mRNAs of MIP-1ß and IL-7. ApN regulation by the ES partly involves p38MAPK.
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Grasa Abdominal/metabolismo , Adipocitos/metabolismo , Adiponectina/biosíntesis , Antagonistas de Receptores de Cannabinoides/farmacología , Obesidad/metabolismo , Epiplón/metabolismo , Piperidinas/farmacología , Pirazoles/farmacología , Receptor Cannabinoide CB1/metabolismo , Grasa Abdominal/efectos de los fármacos , Grasa Abdominal/inmunología , Adipocitos/inmunología , Adipoquinas/biosíntesis , Adiponectina/inmunología , Adiponectina/metabolismo , Tejido Adiposo/efectos de los fármacos , Tejido Adiposo/metabolismo , Adulto , Western Blotting , Células Cultivadas , Quimiocina CCL4/metabolismo , Regulación hacia Abajo , Femenino , Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica , Humanos , Interleucina-1beta/metabolismo , Interleucina-7/genética , Interleucina-7/metabolismo , Masculino , Obesidad/inmunología , Obesidad/cirugía , Epiplón/efectos de los fármacos , Epiplón/inmunología , ARN Mensajero/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Receptor Cannabinoide CB1/efectos de los fármacos , Rimonabant , Inhibidor Tisular de Metaloproteinasa-1/metabolismo , Regulación hacia Arriba , Proteínas Quinasas p38 Activadas por Mitógenos/metabolismoRESUMEN
Obesity plays a causative role in the pathogenesis of the metabolic syndrome. Adipokines may link obesity to its co-morbidities. Most adipokines with pro-inflammatory properties are overproduced with increasing adiposity, while some adipokines with anti-inflammatory or insulin-sensitizing properties, like adiponectin are decreased. This dysregulation of adipokine production may promote obesity-linked metabolic disorders and cardiovascular disease. Besides considering adipokines, this review will also highlight the cellular key players and molecular mechanisms involved in adipose inflammation. Targeting the changes in the cellular composition of adipose tissue, the underlying molecular mechanisms, and the altered production of adipokines may have therapeutic potential in the management of the metabolic syndrome.
Asunto(s)
Adipoquinas/inmunología , Tejido Adiposo , Inflamación/inmunología , Síndrome Metabólico , Adipocitos/citología , Adipocitos/metabolismo , Adiponectina/metabolismo , Tejido Adiposo/inmunología , Tejido Adiposo/patología , Apelina , Distribución de la Grasa Corporal , Quimiocinas/inmunología , Citocinas/inmunología , Citocinas/metabolismo , Humanos , Inflamación/patología , Insulina/metabolismo , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Macrófagos/citología , Macrófagos/inmunología , Síndrome Metabólico/inmunología , Síndrome Metabólico/fisiopatología , Nicotinamida Fosforribosiltransferasa/metabolismo , Obesidad/metabolismo , Transducción de Señal/fisiología , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
Central-omental obesity plays a causative role in the pathogenesis of the metabolic syndrome. Adipokines are involved in the pathogenesis of this syndrome. However, adipokines secreted by omental adipose tissue (OAT) are still poorly characterized in human obesity. Therefore, we searched for novel adipokines abnormally secreted by OAT in obesity and examined their relationships with some features of metabolic syndrome and the respective contribution of adipocytes vs. stromal-vascular cells. OAT from obese and nonobese men was fractionated into adipocytes and SV cells, which were then cultured. Medium was screened by medium-scale protein arrays and ELISAs. Adipokine mRNA levels were measured by real-time RT-qPCR. We detected 16 cytokines secreted by each cellular fraction of lean and obese subjects. Of the 16 cytokines, six adipokines were newly identified as secretory products of OAT, which were dysregulated in obesity: three chemokines (growth-related oncogen factor, RANTES, macrophage inflammatory protein-1beta), one interleukin (IL-7), one tissue inhibitor of metalloproteinases (TIMP-1), and one growth factor (thrombopoietin). Their secretion and expression were enhanced in obesity, with a relatively similar contribution of the two fractions. The higher proportion of macrophages and endothelial cells in obesity may contribute to this enhanced production as well as changes in intrinsic properties of hypertrophied adipocytes. Accordingly, mRNA concentrations of most of these adipokines increased during adipocyte differentiation. Eventually, expression of the investigated adipokines did correlate with several features of the metabolic syndrome. In conclusion, six adipokines were newly identified as oversecreted by OAT in obesity. These adipokines may link obesity to its cardiovascular or metabolic comorbidities.
Asunto(s)
Tejido Adiposo/metabolismo , Citocinas/metabolismo , Síndrome Metabólico/metabolismo , Obesidad/metabolismo , Epiplón/metabolismo , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
AIMS/HYPOTHESIS: Adiponectin is an adipokine that exhibits insulin-sensitising, fat-burning and anti-inflammatory properties as well as modulatory effects on oxidative stress. We examined whether adiponectin could be induced in a non-adipose tissue, skeletal muscle, in response to metabolic or oxidative aggression both in vivo (in a murine model of type 2 diabetes) and in vitro. METHODS: Obese and diabetic ob/ob mice were used and compared with lean littermates. Some obese mice were treated with the antioxidant probucol for 3 weeks. At the end of the experiment, blood was sampled and tibialis anterior muscles were collected for mRNA measurement and immunohistochemistry. Additional in vitro experiments were performed on C2C12 myotubes cultured for up to 48 h. RESULTS: In spite of hypoadiponectinaemia, Adipoq mRNA levels were markedly increased in the skeletal muscle of ob/ob mice and correlated with systemic oxidative stress. Adipoq upregulation was shown in laser-microdissected myocytes of obese mice. Concomitantly, immunoreactivity for adiponectin was enhanced in obese muscle fibres together with lipid infiltration and local markers of oxidative stress. In cultured C2C12 myotubes, a triglyceride mix and reactive oxygen species producers (H2O2 or a lipoperoxidation end-product) upregulated Adipoq expression and adiponectin production. This effect was reversed by an antioxidant. Finally, treatment of obese mice with probucol also attenuated upregulation in muscle. CONCLUSIONS/INTERPRETATION: The paradoxical upregulation of adiponectin in muscle of obese and diabetic mice may result from lipotoxicity and related oxidative stress. This unexpected finding could be viewed as a local protection to counteract ectopic fat deposition and oxidative damage.
Asunto(s)
Adiponectina/biosíntesis , Diabetes Mellitus Tipo 2/fisiopatología , Músculo Esquelético/fisiopatología , Adiponectina/genética , Animales , Antioxidantes/farmacología , Peso Corporal , Células Cultivadas , Cartilla de ADN , Femenino , Regulación de la Expresión Génica , Lípidos/fisiología , Ratones , Ratones Endogámicos C57BL , Ratones Obesos , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/patología , Obesidad , Estrés Oxidativo , Reacción en Cadena de la Polimerasa , Probucol/farmacología , ARN/genética , ARN/aislamiento & purificaciónRESUMEN
Adipose tissue secretes a large number of physiologically active peptides that often share structural properties with cytokines, and are therefore collectively referred to as "adipocytokines". Some of these are almost exclusively secreted by adipose tissue. Leptin, adiponectin and resistin are specific fat-derived hormones that affect fuel homeostasis and insulin action, and may also be involved in hematopoiesis and immune functions. Anorexia nervosa is characterized by chronic self-starvation and severe weight loss, mainly at the expense of adipose tissue. Starvation-induced depletion of fat stores is accompanied by alterations of circulating adipocytokines. Plasma leptin and likely resistin levels are decreased in anorectic patients, while plasma adiponectin levels are increased. These alterations may have potential repercussions in the pathophysiology of anorexia nervosa. Thus, low leptin and high adiponectin may separately or in concert contribute to altered hematopoiesis and immunity, enhanced insulin sensitivity, neuroendocrine disturbances or osteopenia in anorexia nervosa.
Asunto(s)
Adipocitos , Anorexia Nerviosa , Péptidos y Proteínas de Señalización Intercelular , Leptina , Proteínas , Adiponectina , Animales , Hormonas Ectópicas , Humanos , ResistinaRESUMEN
OBJECTIVE: Adiponectin (ApN) is a fat-derived hormone that enhances insulin sensitivity, controls body weight, prevents atherosclerosis and negatively regulates haematopoiesis and immune functions. In contrast to many proteins secreted by adipose tissue, the circulating level of ApN falls in obesity and insulin resistance states. The influence of starvation-induced depletion of fat stores on ApN concentrations is yet unknown. We therefore investigated plasma ApN in anorexia nervosa (AN). PATIENTS AND DESIGN: We measured plasma ApN in 26 female anorectic patients and examined its relationships to several anthropometric or metabolic parameters. Twenty-four age-matched healthy female controls (C) were also studied. RESULTS: Body mass index (BMI) and fat mass were markedly decreased in AN. However, plasma ApN levels were 30% higher in anorectic than in control subjects (P < 0.01), while a reverse pattern was observed for leptin concentrations. When normalized for fatness, ApN values almost doubled in AN. ApN levels were negatively correlated with BMI and fat mass (P < 0.05 in the combined population, AN + C). Insulin sensitivity tended to be 40% higher in AN (n = 7) than in C (n = 12) subjects, and plasma ApN levels were positively correlated with insulin sensitivity (P < 0.05 in AN + C subgroups). Total and low density lipoprotein (LDL)-cholesterol were higher, or tended to be higher, in AN, but there were no correlations between plasma ApN and plasma lipids. By contrast, ApN was related to the lipid profile, in a manner consistent with its antiatherogenic role, in healthy controls [i.e. negatively correlated with triglycerides, total and LDL-cholesterol and total/high density lipoprotein (HDL) cholesterol; P < 0.05 or less for each parameter]. In a multiple regression analysis, BMI and insulin sensitivity in AN were independent determinants for ApN levels, explaining up to approximately 80% of the variance in this measure. CONCLUSIONS: Plasma adiponectin levels are increased in anorexia nervosa. This may, at least in part, be due to the lack of negative feedback exerted by fat mass on adiponectin production and/or to enhanced insulin sensitivity. We speculate that hyperadiponectinaemia could, in turn, contribute to maintain a state of enhanced insulin sensitivity and possibly exacerbate haematological and infectious complications of anorexia nervosa.
Asunto(s)
Anorexia Nerviosa/sangre , Péptidos y Proteínas de Señalización Intercelular , Proteínas/análisis , Adiponectina , Tejido Adiposo/patología , Adulto , Anorexia Nerviosa/patología , Antropometría , Índice de Masa Corporal , Femenino , Humanos , Insulina/sangre , Leptina/sangre , Lípidos/sangre , Análisis de RegresiónRESUMEN
Adiponectin (ApN) is thought to play a major role in the pathogenesis of the Metabolic Syndrome. Production of ApN and regulation of its related gene (apM1) have not yet been studied in human visceral adipose tissue. ApN was mainly associated with adipocyte membranes and abundantly secreted in medium from isolated adipocytes. apM1 gene expression, restricted to the adipocyte fraction of adipose tissue, decreased spontaneously when adipose explants were cultured in basal medium for 24 h while the expression of other adipose genes barely changed (PPARgamma, GAPDH) or increased (PAI-1). Unexpectedly, the fall of apM1 mRNA was prevented by the addition of actinomycin D, an inhibitor of transcription, or cycloheximide, an inhibitor of protein synthesis, and by reducing the amount of adipose tissue cultured per dish, thereby suggesting that a newly synthesized factor released by adipose tissue destabilizes apM1 mRNA. apM1 gene expression was also negatively regulated by glucocorticoids and positively by insulin and IGF-1. This regulation could contribute to the decreased apM1/ApN levels in insulin-resistant patients with obesity and the Metabolic Syndrome.
Asunto(s)
Tejido Adiposo/metabolismo , Péptidos y Proteínas de Señalización Intercelular , Obesidad/metabolismo , Biosíntesis de Proteínas , Proteínas/metabolismo , Adipocitos/efectos de los fármacos , Adipocitos/metabolismo , Adiponectina , Células Cultivadas , Técnicas de Cultivo , Dactinomicina/farmacología , Dexametasona/farmacología , Femenino , Regulación de la Expresión Génica , Glucocorticoides/farmacología , Humanos , Insulina/farmacología , Factor I del Crecimiento Similar a la Insulina/farmacología , Masculino , Persona de Mediana Edad , Inhibidores de la Síntesis del Ácido Nucleico/farmacología , Obesidad/genética , Proteínas/genética , Estabilidad del ARN , ARN Mensajero/biosíntesis , VíscerasRESUMEN
The syntheses and the solid state structural and spectroscopic solution characterizations of VO(Me-acac)2 and VO(Et-acac)2 (where Me-acac is 3-methyl-2,4-pentanedionato and Et-acac is 3-ethyl-2,4-pentanedionato) have been conducted since both VO(acac)2 and VO(Et-acac)2 have long-term in vivo insulin-mimetic effects in streptozotocin-induced diabetic Wistar rats. X-ray structural characterizations of VO(Me-acac)2 and VO(Et-acac)2 show that both contain five-coordinate vanadium similar to the parent VO(acac)2. The unit cells for VO(Et-acac)2 and VO(Me-acac)2 are both triclinic, P1, with a = 9.29970(10) A, b = 13.6117(2) A, c = 13.6642(2) A, alpha = 94.1770(10) degrees, beta = 106.4770(10) degrees, gamma = 106.6350(10) degrees for VO(Et-acac)2 and a = 7.72969(4) A, b = 8.1856(5) A, c = 11.9029(6) A, alpha = 79.927(2) degrees, beta = 73.988(2)degrees, gamma = 65.1790(10)degrees for VO(Me-acac)2. The total concentration of EPR-observable vanadium(IV) species for VO(acac)2 and derivatives in water solution at 20 degreesC was determined by double integration of the EPR spectra and apportioned between individual species on the basis of computer simulations of the spectra. Three species were observed, and the concentrations were found to be time, pH, temperature, and salt dependent. The three complexes are assigned as the trans-VO(acac)2.H2O adduct, cis-VO(acac)2.H2O adduct, and a hydrolysis product containing one vanadium atom and one R-acac- group. The reaction rate for conversion of species was slower for VO(acac)2 than for VO(malto)2, VO(Et-acac)2, and VO(Me-acac)2; however, in aqueous solution the rates for all of these species are slow compared to those of other vanadium species. The concentration of vanadium(V) species was determined by 51V NMR. The visible spectra were time dependent, consistent with the changes in species concentrations that were observed in the EPR and NMR spectra. EPR and visible spectroscopic studies of solutions prepared as for administration to diabetic rats documented both a salt effect on speciation and formation of a new halogen-containing complex. Compound efficacy with respect to long-term lowering of plasma glucose levels in diabetic rats traces the concentration of the hydrolysis product in the administration solution.
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Insulina/fisiología , Compuestos Organometálicos/química , Vanadatos/química , Animales , Glucemia/efectos de los fármacos , Cristalografía por Rayos X , Diabetes Mellitus Experimental/inducido químicamente , Modelos Animales de Enfermedad , Espectroscopía de Resonancia por Spin del Electrón , Insulina/química , Espectroscopía de Resonancia Magnética , Masculino , Imitación Molecular , Estructura Molecular , Compuestos Organometálicos/farmacología , Compuestos Organometálicos/uso terapéutico , Ratas , Ratas Wistar , Estreptozocina , Relación Estructura-Actividad , Vanadatos/farmacología , Vanadatos/uso terapéuticoRESUMEN
Plasma levels of type 1 plasminogen activator inhibitor (PAI-1), a risk factor for cardiovascular disease, are elevated in obese subjects, especially those with omental fat accumulation. We investigated the hormonal control of PAI-1 gene expression and secretion in cultured human adipose tissue. We more particularly focused on the effects of glucocorticoids, insulin, cAMP, and catecholamines in explants from the omental region. The addition of dexamethasone to the culture medium increased PAI-1 secretion in a time-dependent manner for up to 24 h. The stimulation by the glucocorticoid was preceded by a 2-fold rise in PAI-1 messenger ribonucleic acid levels between 4-8 h of culture. The effectiveness of the glucocorticoid was concentration dependent, with a half-maximal effect within a physiological range. This stimulation was also observed in sc fat, but dexamethasone-stimulated as well as basal PAI-1 secretion rates were always higher in omental fat. Unlike dexamethasone, 24-h insulin did not modify PAI-1 secretion while accelerating glucose consumption. In contrast, 24-h cAMP inhibited PAI-1 gene expression and protein production under basal conditions and in the presence of dexamethasone. This inhibition was already detectable after 1 h and was maximal after 4 h at the level of gene expression. It occurred in both omental and sc adipose tissues. Importantly, epinephrine dose dependently inhibited PAI-1 parameters, an effect that was reproduced by isoproterenol. Dexamethasone- and cAMP-induced changes in PAI-1 messenger ribonucleic acid abundance were similar in explants and isolated fat cells. In isolated stromal-vascular cells, only dexamethasone was effective. In conclusion, we provide evidence for a reciprocal regulation of PAI-1 by dexamethasone (positive effector) and cAMP/catecholamines (negative effectors) in cultured human adipose tissue. The stimulation by glucocorticoids could contribute to enhanced production of PAI-1 by adipose tissue and high plasma levels of PAI-1 associated with central obesity and thereby be a link between this disorder and cardiovascular disease. Impaired inhibition by catecholamines could also contribute, as in vivo adipose tissue responses to these hormones are usually blunted in obese individuals.
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Tejido Adiposo/metabolismo , Catecolaminas/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Glucocorticoides/farmacología , Inhibidor 1 de Activador Plasminogénico/genética , Agonistas Adrenérgicos beta/farmacología , AMP Cíclico/análogos & derivados , AMP Cíclico/farmacología , Dexametasona/farmacología , Relación Dosis-Respuesta a Droga , Epinefrina/farmacología , Femenino , Humanos , Insulina/farmacología , Isoproterenol/farmacología , Cinética , Masculino , Persona de Mediana Edad , Obesidad/metabolismo , ARN Mensajero/metabolismoRESUMEN
1. Vanadium compounds can mimic actions of insulin through alternative signalling pathways. The effects of three organic vanadium compounds were studied in non-ketotic, streptozotocin-diabetic rats: vanadyl acetylacetonate (VAc), vanadyl 3-ethylacetylacetonate (VEt), and bis(maltolato)oxovanadium (VM). A simple inorganic vanadium salt, vanadyl sulphate (VS) was also studied. 2. Oral administration of the three organic vanadium compounds (125 mg vanadium element 1(-1) in drinking fluids) for up to 3 months induced a faster and larger fall in glycemia (VAc being the most potent) than VS. Glucosuria and tolerance to a glucose load were improved accordingly. 3. Activities and mRNA levels of key glycolytic enzymes (glucokinase and L-type pyruvate kinase) which are suppressed in the diabetic liver, were restored by vanadium treatment. The organic forms showed greater efficacy than VS, especially VAc. 4. VAc rats exhibited the highest levels of plasma or tissue vanadium, most likely due to a greater intestinal absorption. However, VAc retained its potency when given as a single i.p. injection to diabetic rats. Moreover, there was no relationship between plasma or tissue vanadium levels and any parameters of glucose homeostasis and hepatic glucose metabolism. Thus, these data suggest that differences in potency between compounds are due to differences in their insulin-like properties. 5. There was no marked toxicity observed on hepatic or renal function. However, diarrhoea occurred in 50% of rats chronically treated with VS, but not in those receiving the organic compounds. 6. In conclusion, organic vanadium compounds, in particular VAc, correct the hyperglycemia and impaired hepatic glycolysis of diabetic rats more safely and potently than VS. This is not simply due to improved intestinal absorption, indicating more potent insulin-like properties.
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Glucosa/metabolismo , Ligandos , Compuestos de Vanadio/farmacología , Administración Oral , Animales , Glucemia/efectos de los fármacos , Glucemia/metabolismo , Peso Corporal/efectos de los fármacos , Diabetes Mellitus Experimental/metabolismo , Desinfectantes/farmacología , Glucoquinasa/efectos de los fármacos , Glucoquinasa/genética , Glucoquinasa/metabolismo , Hidroxibutiratos/química , Hidroxibutiratos/farmacología , Hipoglucemiantes/farmacología , Inyecciones Intraperitoneales , Insulina/metabolismo , Islotes Pancreáticos/efectos de los fármacos , Islotes Pancreáticos/metabolismo , Hígado/efectos de los fármacos , Hígado/metabolismo , Glucógeno Hepático/metabolismo , Masculino , Músculos/efectos de los fármacos , Músculos/metabolismo , Compuestos Organometálicos/farmacología , Pentanonas/química , Pentanonas/farmacología , Fosfoenolpiruvato Carboxiquinasa (GTP)/efectos de los fármacos , Fosfoenolpiruvato Carboxiquinasa (GTP)/genética , Fosfoenolpiruvato Carboxiquinasa (GTP)/metabolismo , Pironas/química , Pironas/farmacología , Piruvato Quinasa/efectos de los fármacos , Piruvato Quinasa/genética , Piruvato Quinasa/metabolismo , ARN Mensajero/efectos de los fármacos , ARN Mensajero/genética , Ratas , Ratas Wistar , Factores de Tiempo , Vanadatos/química , Vanadatos/farmacología , Compuestos de Vanadio/químicaRESUMEN
Leptin is a protein encoded by the ob gene that is expressed in adipocytes and regulates eating behavior via neuroendocrine mechanisms. Plasma leptin levels have been shown to correlate with weight and body fat in normal, obese and anorexic subjects. In the last of these populations, the dynamic profile of plasma leptin levels during short-term refeeding has never been assessed. We thus investigated basal plasma leptin levels in 29 female patients with anorexia nervosa (AN) (age 21.9 +/- 1.4 years, body mass index (BMI) 15.2 +/- 0.3 kg/m2) and in 80 normal female controls (age 21.2 +/- 0.2 years, BMI 20.3 +/- 0.3 kg/m2, mean +/- S.E.M.). Basal plasma leptin levels in AN were decreased by 77% compared with controls (2.5 +/- 0.2 vs 11.1 +/- 0.7 ng/ml, P < 0.0001). In both AN subjects and controls, plasma leptin levels correlated significantly with BMI (r2 = 0.448, P < 0.0001 and r2 = 0.339, P < 0.0001 respectively). Five AN patients (four female, one male, age 22.0 +/- 4.7 years, BMI 14.2 +/- 0.4 kg/m2, body fat 4.3 +/- 0.9 kg or 11.0 +/- 1.9% of body weight, basal metabolic rate (BMR) 958 +/- 122 kcal/day) were studied during a 3-day refeeding period and compared with eight control subjects (two male, six female, age 25.7 +/- 1.2 years, BMI 21.3 +/- 0.8 kg/m2, body fat 15.1 +/- 0.9 kg or 24.6 +/- 1.7%, BMR 1455 +/- 78 kcal/day) submitted to 36-h fasting. The amount of calories administered was based on BMR + 20% (carbohydrate 60%, protein 17%, fat 23%). In contrast to the rise in leptin levels that occurred during refeeding after a prolonged fast period in normal subjects, plasma leptin levels remained low and unchanged throughout the 3 days of renutrition in AN patients. The circadian rhythm of leptin was also completely abolished. This contrasted with the preserved circadian variations of cortisol, whose mean levels were increased. In conclusion, we confirmed that plasma leptin levels are low in AN and correlate with body weight. We further demonstrated that plasma leptin levels do not respond to short-term refeeding in anorexic patients in whom circadian variations are not restored, which suggests that the acute regulation of leptin by positive changes in energy balance is not preserved under a critical threshold of body fat.
Asunto(s)
Anorexia Nerviosa/sangre , Ritmo Circadiano/fisiología , Proteínas/metabolismo , Adulto , Análisis de Varianza , Anorexia Nerviosa/dietoterapia , Estudios de Casos y Controles , Femenino , Humanos , Hidrocortisona/metabolismo , Leptina , Masculino , Tasa de Secreción , Tirotropina/metabolismoRESUMEN
The direct role of hormones on leptin synthesis has not yet been studied in cultured adipose cells or tissue from lean and obese subjects. Moreover, this hormonal regulation has never been addressed in human visceral fat, although this site plays a determinant role in obesity-linked disorders. In this study, we investigated the hormonal control of ob expression and leptin production in cultured visceral adipose tissue from lean and obese subjects. We more particularly focused on the interactions between glucocorticoids and insulin. We also briefly tackled the role of cAMP, which is still unknown in man. Visceral (and subcutaneous) adipose tissues from eight obese (body mass index, 41 +/- 2 kg/m2) and nine nonobese (24 +/- 1 kg/m2) subjects were sampled during elective abdominal surgery, and explants were cultured for up to 48 h in MEM. The addition of dexamethasone to the medium increased ob gene expression and leptin secretion in a time-dependent manner. Forty-eight hours after dexamethasone (50 nmol/L) addition, the cumulative integrated ob messenger ribonucleic acid (mRNA) and leptin responses were, respectively, approximately 5- and 4-fold higher in obese than in lean subjects. These responses closely correlated with the body mass index. The stimulatory effect of the glucocorticoid was also concentration dependent (EC50 = approximately 10 nmol/L). Although the maximal response was higher in obese than in lean subjects, the EC50 values were roughly similar in both groups. Unlike dexamethasone, insulin had no direct stimulatory effect on ob gene expression and leptin secretion. Singularly, insulin even inhibited the dexamethasone-induced rise in ob mRNA and leptin release. This inhibition was observed in both lean and obese subjects, whereas the expected stimulation of insulin on glucose metabolism and the accumulation of mRNA species for the insulin-sensitive transporter GLUT4 and glyceraldehyde-3-phosphate dehydrogenase occurred in lean patients only. This inhibitory effect was already detectable at 10 nmol/L insulin and was also observed in subcutaneous fat. Although a lowering of intracellular cAMP concentrations is involved in some of the effects of insulin on adipose tissue, this cannot account for the present finding, because the addition of cAMP to the medium also decreased ob mRNA and leptin secretion (regardless of whether dexamethasone was present). In conclusion, glucocorticoids, at physiological concentrations, stimulated leptin secretion by enhancing the pretranslational machinery in human visceral fat. This effect was more pronounced in obese subjects due to a greater responsiveness of the ob gene and could contribute to the metabolic abnormalities associated with central obesity by para/endocrine actions of hyperleptinemia on adipocytes and liver. Unlike dexamethasone, insulin had no direct stimulatory effect on ob gene expression and leptin secretion, and even prevented the positive response to dexamethasone by a cAMP-independent mechanism that remained functional despite insulin resistance.
Asunto(s)
Tejido Adiposo/fisiopatología , Regulación de la Expresión Génica/fisiología , Hormonas/fisiología , Obesidad/genética , Proteínas/metabolismo , Tejido Adiposo/efectos de los fármacos , Adulto , Técnicas de Cultivo , AMP Cíclico/farmacología , Dexametasona/farmacología , Femenino , Regulación de la Expresión Génica/efectos de los fármacos , Glucocorticoides/farmacología , Humanos , Insulina/farmacología , Leptina , Masculino , Persona de Mediana Edad , Obesidad/metabolismo , ARN Mensajero/metabolismo , Valores de Referencia , VíscerasRESUMEN
Molybdate (Mo) exerts insulinomimetic effects in vitro. In this study, we evaluated whether Mo can improve glucose homeostasis in genetically obese, insulin-resistant ob/ob mice. Oral administration of Mo (174 mg/kg molybdenum element) for 7 weeks did not affect body weight, but decreased the hyperglycaemia (approximately 20 mM) of obese mice to the levels of lean (L) (+/+) mice, and reduced the hyperinsulinaemia to one-sixth of pretreatment levels. Tolerance to oral glucose was improved: total glucose area was 30% lower in Mo-treated mice than in untreated ob/ob mice (O), while the total insulin area was halved. Hepatic glucokinase (GK) mRNA level and activity were unchanged in O mice compared with L mice, but the mRNA level and activity of L-type pyruvate kinase (L-PK) were increased in O mice by 3.5- and 1.7-fold respectively. Mo treatment increased GK mRNA levels and activity (by approximately 2.2-fold and 61% compared with O values), and had no, or only a mild, effect on the already increased L-PK variables. mRNA levels and activity of the gluconeogenic enzyme, phosphoenolpyruvate carboxykinase (PEPCK) were augmented in O liver (sixfold and by 57% respectively), and these were reduced by Mo treatment. Insulin binding to partially purified receptors from liver was reduced in O mice and restored by Mo treatment. Despite this correction, overall receptor tyrosine kinase activity was not improved in Mo mice. Moreover, the overexpression (by two- to fourfold) of the cytokine tumour necrosis factor alpha (TNF alpha) in white adipose tissue, which may have a determinant role in the insulin resistance of the O mice, was unaffected by Mo. Likewise, overexpression of the ob gene in white adipose tissue was unchanged by Mo. In conclusion, Mo markedly improved glucose homeostasis in the ob/ob mice by an insulin-like action which appeared to be exerted distal to the insulin receptor tyrosine kinase step. The blood glucose-lowering effect of Mo was unrelated to over-expression of the TNF alpha and ob genes in O mice, but resulted at least in part from attenuation of liver insulin resistance by the reversal of pre-translational regulatory defects in these mice.
Asunto(s)
Glucemia/metabolismo , Insulina/metabolismo , Hígado/metabolismo , Molibdeno/farmacología , Obesidad/metabolismo , Tejido Adiposo/efectos de los fármacos , Tejido Adiposo/metabolismo , Administración Oral , Animales , Femenino , Expresión Génica/efectos de los fármacos , Glucoquinasa/genética , Glucoquinasa/metabolismo , Homeostasis , Resistencia a la Insulina/fisiología , Leptina , Masculino , Ratones , Ratones Mutantes , Proteínas/genética , Piruvato Quinasa/genética , Piruvato Quinasa/metabolismo , Factor de Necrosis Tumoral alfa/genéticaRESUMEN
The ob gene, specifically expressed in fat cells, encodes leptin, a hormone that induces satiety and increases energy expenditure. In this study, we investigated the interactions between glucocorticoids and insulin on ob gene expression in cultured explants of rat adipose tissue. Only low levels of ob mRNA were detected when adipose tissue from fasted rats was cultured for 12-24 h in minimal essential medium. However, the addition of dexamethasone to the medium increased ob gene expression in a concentration-dependent manner (EC50 10 nM). With 1 microM dexamethasone, ob mRNA levels were similar to those in fresh fat pads from fed rats, reaching a maximum after 12 h. The effect of dexamethasone was blocked by actinomycin D, which indicates an action on transcription. This effect was increased when a minimum amount of fuel (glucose or a mixture of lactate and pyruvate) was supplied in the medium. Unlike dexamethasone, insulin, even when combined with high glucose concentrations, did not induce ob expression, although it strongly increased the accumulation of mRNA species for fatty acid synthase (FAS), the insulin-sensitive glucose transporter GLUT4 and the gamma isoform of peroxisome proliferator-activated receptor (PPARgamma). Unexpectedly, insulin dose-dependently inhibited dexamethasone-induced ob mRNA accumulation. This effect was observed at low concentrations of insulin (IC50 1 nM) and was delayed in onset, beginning after 6-9 h of culture. It was mimicked by insulin-like growth factor 1 (IGF-1) (100 nM). The inhibition by insulin was only detectable when fuels were present and/or when a critical level of ob expression was reached. As this inhibitory effect was reversed by cycloheximide, this suggests that it required ongoing protein synthesis. In conclusion, unlike dexamethasone, insulin had no direct stimulatory effect on ob gene expression. On the other hand, insulin (and IGF-1) even inhibited the dexamethasone-induced accumulation of ob mRNA. The underlying mechanism involved ongoing synthesis of an inhibitory protein by insulin, which is in keeping with its delayed effect. Moreover, the expression of genes for FAS, GLUT4 and PPARgamma may be inversely related to that of ob.
Asunto(s)
Tejido Adiposo/efectos de los fármacos , Dexametasona/antagonistas & inhibidores , Regulación de la Expresión Génica/efectos de los fármacos , Factor I del Crecimiento Similar a la Insulina/farmacología , Insulina/farmacología , Proteínas Musculares , Biosíntesis de Proteínas , 1-Metil-3-Isobutilxantina/farmacología , Tejido Adiposo/metabolismo , Animales , AMP Cíclico/farmacología , Cicloheximida/farmacología , Dexametasona/farmacología , Metabolismo Energético , Inducción Enzimática/efectos de los fármacos , Epidídimo , Ácido Graso Sintasas/biosíntesis , Ácido Graso Sintasas/genética , Ácidos Grasos/farmacología , Glucosa/farmacología , Transportador de Glucosa de Tipo 4 , Ácido Láctico/farmacología , Leptina , Masculino , Microcuerpos/enzimología , Proteínas de Transporte de Monosacáridos/biosíntesis , Proteínas de Transporte de Monosacáridos/genética , Técnicas de Cultivo de Órganos , Inhibidores de la Síntesis de la Proteína/farmacología , Proteínas/genética , Ácido Pirúvico/farmacología , ARN Mensajero/biosíntesis , Ratas , Ratas Wistar , Receptores Citoplasmáticos y Nucleares/biosíntesis , Receptores Citoplasmáticos y Nucleares/genética , Transducción de Señal/efectos de los fármacos , Acetato de Tetradecanoilforbol/farmacología , Factores de Transcripción/biosíntesis , Factores de Transcripción/genéticaRESUMEN
The ob gene encodes leptin, a hormone which induces satiety and increases energy expenditure. The peroxisome proliferator-activated receptor gamma 2 isoform (PPAR gamma 2) gene encodes a transcription factor which controls adipocyte differentiation and expression of fat-specific genes. We have studied the regulation of these two genes in white adipose tissue (WAT) during the suckling-weaning transition. Suckling rats ingest a high-fat diet (milk). Fat-pad weight barely varied during the last week of suckling. ob mRNA levels, which were very low in 15-day-old rats, rose approximately 6-fold until weaning at 21 days. When the rats were weaned on to a standard (high-carbohydrate) laboratory chow, epididymal WAT enlarged approximately 7-fold, and ob mRNA kept increasing progressively and doubled between 21 and 30 days. This evolution contrasted with that of fatty acid synthase (FAS) mRNA, which increased sharply, but only after weaning. To distinguish between the influence of developmental and nutritional factors on ob expression, a group of rats was weaned on to a high-fat diet. This prevented the rise in glycaemia and insulinaemia and the decrease in plasma non-esterified fatty acids which otherwise occurred at weaning. This also resulted in a slight (10-15%) decrease in food intake and body weight gain. Under this high-fat diet, the rise of ob mRNA in WAT was augmented (3.7-fold in 30- versus 21-day-old pups), whereas the normal rise in FAS mRNA levels was attenuated. Fat-pad weights and adipocyte cell size and number were roughly similar in high-carbohydrate- and high-fat-weaned pups. mRNA levels of PPAR gamma 2, like those of ob, were low in the WAT of 15-day-old suckling pups, doubled at 21 days, and reached a maximum as soon as 23 days. This evolution further differed from that of ob mRNA in not being influenced by diet composition. In conclusion, ob expression markedly increases during the suckling-weaning transition, and this effect is accentuated by a high-fat diet. Qualitative nutritional changes in ob mRNA were correlated with neither acute changes in adipose-tissue mass, nor cell size/number, nor variations in insulinaemia. PPAR gamma 2 also increased during suckling, but rapidly reached a plateau after weaning and no longer changed thereafter. Unlike ob, PPAR gamma 2 was not influenced by the diet composition.
Asunto(s)
Tejido Adiposo/metabolismo , Regulación del Desarrollo de la Expresión Génica , Microcuerpos/metabolismo , Obesidad/genética , Receptores Citoplasmáticos y Nucleares/biosíntesis , Receptores Citoplasmáticos y Nucleares/genética , Factores de Transcripción/biosíntesis , Factores de Transcripción/genética , Tejido Adiposo/crecimiento & desarrollo , Animales , Animales Lactantes/genética , Animales Lactantes/crecimiento & desarrollo , Glucemia/metabolismo , Peso Corporal/genética , Masculino , Ratas , Ratas Wistar , DesteteRESUMEN
Molybdenum mimics certain insulin actions in vitro. We have investigated the effects of oral administration of Na2MoO4 (Mo) for 8 wk on carbohydrate and lipid metabolism in streptozotocin-diabetic rats. Mo decreased hyperglycemia and glucosuria by 75% and corrected the elevation of plasma nonesterified fatty acids. Tolerance to glucose loads was improved, and glycogen stores were replenished. These effects were not due to a rise of insulinemia. In liver, Mo restored the blunted mRNA and activity of glucokinase and pyruvate kinase and decreased to normal phosphoenolpyruvate carboxykinase values. Finally, Mo totally reversed the low expression and activity of acetyl-CoA carboxylase and fatty acid synthase in liver, but not in white adipose tissue. In conclusion, Mo exerts a marked blood glucose-lowering effect in diabetic rats by an insulin-like action. This effect results in part from a restoration of hepatic glucose metabolism and is associated with a tissue-specific correction of lipogenic enzyme gene expression, both processes being essentially mediated by reversal of impaired pretranslational regulatory mechanisms. These observations raise new therapeutic perspectives in diabetes, particularly in the insulin-resistant condition.
Asunto(s)
Diabetes Mellitus Experimental/metabolismo , Glucosa/metabolismo , Metabolismo de los Lípidos , Molibdeno/farmacología , Animales , Glucemia/análisis , Diabetes Mellitus Experimental/fisiopatología , Enzimas/genética , Prueba de Tolerancia a la Glucosa , Homeostasis , Hígado/metabolismo , Masculino , ARN Mensajero/metabolismo , Ratas , Ratas WistarRESUMEN
Selenium is a trace element that exerts certain insulin-like actions in vitro. In this study, we evaluated its in vivo effects on the glucose homeostasis of rats made diabetic and insulin-deficient by streptozotocin. Na2SeO4 was administered ad libitum in drinking water and/or food for 10 weeks. The elevated plasma glucose levels (approximately 25 mmol/l) and glucosuria (approximately 85 mmol/day) of untreated rats were decreased by 50 and 80%, respectively, by selenate treatment. The beneficial effect of selenate was also evident during oral and intravenous glucose tolerance tests: the integrated glucose responses were decreased by 40-50% as compared to those in untreated rats. These effects were not due to an increase in plasma insulin levels. Compared to non-diabetic rats, pancreatic insulin reserves were reduced by more than 90% in treated and untreated diabetic rats. The hepatic activities and mRNA levels of two key glycolytic enzymes, glucokinase and L-type pyruvate kinase were blunted in diabetic rats. They increased approximately two- to threefold after selenate treatment, to reach 40-75% of the values in non-diabetic rats. In contrast, elevated activity and mRNA levels of the gluconeogenic enzyme, phosphoenolpyruvate carboxykinase, were reduced by 40-65% after selenate administration. Since selenate induced a moderate decrease in body weight due to an anorexigenic effect, we checked that there was no improvement of glucose homeostasis or hepatic glucose metabolism in an additional group of calorie-restricted diabetic rats, which was weight-matched with the selenate group. In addition, no obvious toxic side-effects on the kidney or liver were observed in the rats receiving selenate. In conclusion, selenate induces a sustained improvement of glucose homeostasis in streptozotocin-diabetic rats by an insulin-like action, which involves partial correction of altered pretranslational regulatory mechanisms in liver metabolism.
Asunto(s)
Glucemia/metabolismo , Diabetes Mellitus Experimental/metabolismo , Glucosa/metabolismo , Hígado/enzimología , Compuestos de Selenio/farmacología , Administración Oral , Animales , Glucemia/efectos de los fármacos , Northern Blotting , Peso Corporal/efectos de los fármacos , Sondas de ADN , Diabetes Mellitus Experimental/sangre , Diabetes Mellitus Experimental/tratamiento farmacológico , Expresión Génica/efectos de los fármacos , Glucoquinasa/biosíntesis , Gluconeogénesis , Prueba de Tolerancia a la Glucosa , Glucólisis , Homeostasis/efectos de los fármacos , Insulina/sangre , Islotes Pancreáticos/efectos de los fármacos , Islotes Pancreáticos/fisiología , Islotes Pancreáticos/fisiopatología , Hígado/efectos de los fármacos , Masculino , Fosfoenolpiruvato Carboxiquinasa (GTP)/biosíntesis , Piruvato Quinasa/biosíntesis , Ratas , Ratas Wistar , Ácido Selénico , Selenio/metabolismo , Compuestos de Selenio/administración & dosificaciónRESUMEN
Vanadium is a trace element that has raised increasing interest in diabetology since the discovery of its insulin-like properties in vitro and in vivo. This brief article reviews the most recent data concerning the beneficial effects of vanadium compounds on fuel homeostasis in animal models of insulinopenic (Type I) or insulin-resistant (Type II) diabetes. These studies open obvious therapeutic possibilities in diabetes, and more particularly, in states of insulin resistance.
Asunto(s)
Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 1/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Vanadatos/farmacología , Animales , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Tipo 1/genética , Diabetes Mellitus Tipo 2/genética , Expresión Génica/efectos de los fármacos , Homeostasis/efectos de los fármacos , Homeostasis/genéticaRESUMEN
ob gene regulation is as yet unknown. We first examined whether the ob gene is under physiological control by the nutritional state. Fasting produced a sharp (95%) decrease of ob mRNA in epididymal and inguinal fat pads from 24 h onward. Refeeding rapidly (3-6 h) re-induced ob gene expression and corrected it within 24 h. Similar changes in fatty acid synthase (FAS) and GLUT4 mRNAs were observed, whereas phosphoenolpyruvate carboxykinase (PEPCK) mRNA showed an opposite evolution. We next examined the potential role of insulin. In adipose tissue of streptozotocin-diabetic rats, ob mRNA levels were decreased by 80%. Insulin treatment (4 days) only marginally increased ob mRNA, but restored euglycemia and overcorrected FAS, GLUT4 and PEPCK expression. In conclusion, we provide evidence for a physiological regulation of ob gene by variations in the nutritional state. We also show that ob expression is impaired in streptozotocin-diabetic rats and only slightly restored by insulin treatment, which suggests that ob gene is not or only minimally regulated by the hormone.