RESUMO
Stanniocalcin-1 and -2 belong to a family of molecules that exhibit both paracrine and autocrine effects in mammalian cells. Human stanniocalcin-1 (hSTC-1) is expressed in a wide range of tissues, including white adipose tissue. In fed rats, hSTC-1 increases carbon flux from glucose to lipids in retroperitoneal white adipose tissue. Human stanniocalcin-2 (hSTC-2) is expressed in almost all tissues and regulates various biological processes. The aim of this work was to study the action of hSTC-1 and hSTC-2 in the lipid and glucose metabolism of epididymal white adipose tissue (eWAT) in rats in different nutritional states. This study shows for the first time an opposite effect of hSTC-1 and hSTC-2 on glyceride-glycerol generation from glucose in eWAT of fed rats. hSTC-1 stimulated the storage of triacylglycerol in eWAT in the postprandial period, increasing glucose uptake and glyceride-glycerol generation from 14C-glucose. hSTC-2 decreased triacylglycerol synthesis, reducing glyceride-glycerol generation from 14C-glucose, direct phosphorylation of glycerol, and fatty acid synthesis from 14C-glucose in eWAT of fed rats. However, both hormones increased glucose uptake in fed and fasting states. These findings provide evidence for a direct role of hSTC-1 and hSTC-2 in the regulation of lipid and glucose metabolism in eWAT of rats.
Assuntos
Tecido Adiposo Branco/metabolismo , Glucose/metabolismo , Glicoproteínas/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Metabolismo dos Lipídeos , Animais , Epididimo/metabolismo , Jejum/fisiologia , Masculino , Período Pós-Prandial/fisiologia , Ratos , Ratos Wistar , Triglicerídeos/biossínteseRESUMO
The present work assesses in vitro the role of human Stanniocalcin 1 (hSTC-1) in glucose metabolism in white retroperitoneal adipose tissue (WRAT) from fed rat. In the fed state, hSTC1 increases the incorporation of 14C from glucose into lipids in the rat WRAT. The increase in lipogenesis capacity supports the hypothesis that the activity of the glycerol-3-phosphate-generating pathway (glycolysis) from glucose is regulated by hSTC-1. The effect of hSTC-1 on de novo fatty acid synthesis and on glucose oxidation in WRAT is supported by an 85 % increase in 14CO2 production from 14C-glucose. The incubation of WRAT in the presence of hSTC-1 maintained the ADP/ATP ratio close to the control group. The presence of hSTC-1 in the incubation medium did not inhibit the lipolytic effect of epinephrine. In conclusion, hSTC-1 is one of the hormonal factors that control glucose metabolism in WRAT in the fed state.
Assuntos
Tecido Adiposo Branco/metabolismo , Glucose/metabolismo , Glicoproteínas/metabolismo , Metabolismo dos Lipídeos , Trifosfato de Adenosina/metabolismo , Animais , Dióxido de Carbono/metabolismo , Ácidos Graxos/metabolismo , Humanos , Lipogênese , Masculino , Oxirredução , Ratos , Ratos WistarRESUMO
The present study assesses the effects of osmotic stress on phosphoenolpyruvate carboxykinase (PEPCK), fructose 1,6-bisphosphatase (FBPase) and glucose 6-phosphatase (G6Pase) activities and (14)C-total lipid synthesis from (14)C-glycine in the anterior and posterior gills, jaw muscle, and hepatopancreas of Neohelice granulata. In posterior gills, 24-h exposure to hyperosmotic stress increased PEPCK, FBPase and G6Pase activities. Increase in (14)C-lipid synthesis was associated to the decrease in PEPCK activity after 72-h exposure to hyperosmotic stress. Hypo-osmotic stress decreased PEPCK and G6Pase activities in posterior gills; however, (14)C-lipids increased after 72-h exposure to stress. In anterior gills, decreases in the G6Pase activity after 72-h of hyperosmotic stress and in (14)C-lipogenesis after 144-h were observed, while PEPCK activity increased after 144 h. Exposure to hypo-osmotic stress increased (14)C-lipid synthesis and PEPCK activity in anterior gills. Muscle G6Pase activity increased after 72-h exposure to hypo-osmotic stress; however, no significant change was observed in the lipogenesis. PEPCK decreased in muscle after 144-h exposure to hyperosmotic, coinciding with increased (14)C-lipid synthesis. In the hepatopancreas, a decrease in the (14)C-lipogenesis occurred after 24-h exposure to hyperosmotic stress, accompanied by increase in (14)C-lipid synthesis. Additionally, PEPCK activity returned to control levels. The hepatopancreatic lipogenesis from amino acids was not involved in the metabolic adjustment during hypo-osmotic stress. However, gluconeogenesis is one of the pathways involved in the adjustment of the intracellular concentration of nitrogenated compounds.
Assuntos
Braquiúros/enzimologia , Gluconeogênese , Lipídeos/biossíntese , Animais , Carboxiliases/metabolismo , Frutose-Bifosfatase/metabolismo , Brânquias/enzimologia , Glucose-6-Fosfatase/metabolismo , Hepatopâncreas/enzimologia , Masculino , Músculos/enzimologia , Pressão OsmóticaRESUMO
The ketogenic diet (KD), characterized by high fat and low carbohydrate and protein contents, has been proposed to be beneficial in children with epilepsy disorders not helped by conventional anti-epileptic drug treatment. Weight loss and inadequate growth is an important drawback of this diet and metabolic causes are not well characterized. The aim of this study was to examine body weight variation during KD feeding for 6 wk of Wistar rats; fat mass and adipocyte cytosolic phosphoenolpyruvate carboxykinase (PEPCK) activity were also observed. PEPCK activity was determined based on the [H(14)CO(3) (-)]-oxaloacetate exchange reaction. KD-fed rats gained weight at a less rapid rate than normal-fed rats, but with a significant increment in fat mass. The fat mass/body weight ratio already differed between ketogenic and control rats after the first week of treatment, and was 2.4 x higher in ketogenic rats. The visceral lipogenesis was supported by an increment in adipocyte PEPCK, aiming to provide glycerol 3-phosphate to triacylglycerol synthesis and this fat accumulation was accompanied by glucose intolerance. These data contribute to our understanding of the metabolic effects of the KD in adipose tissue and liver and suggest some potential risks of this diet, particularly visceral fat accumulation.
Assuntos
Tecido Adiposo/anatomia & histologia , Dieta Cetogênica/estatística & dados numéricos , Fosfoenolpiruvato Carboxiquinase (ATP)/metabolismo , Redução de Peso/fisiologia , Tecido Adiposo/efeitos dos fármacos , Animais , Criança , Colesterol/sangue , Epilepsia/prevenção & controle , Humanos , Masculino , Ratos , Ratos Wistar , Triglicerídeos/sangue , Redução de Peso/efeitos dos fármacosRESUMO
BACKGROUND: Uterine leiomyomas are the commonest tumors of the genital tract. Growth factors seem to be implicated in the development of leiomyoma. OBJECTIVE: To determine the insulin receptor (IR) tyrosine kinase activity--phosphorylation of exogenous substrate poly(Glu 4: Tyr 1)--and insulin receptor substrate 1 expression in normal myometrium and leiomyoma. STUDY DESIGN: The study group consisted of 12 women with leiomyoma undergoing routine hysterectomy. Samples of leiomyoma and adjacent normal myometrium were obtained at the time of operation. Plasma membrane fractions were prepared and samples were incubated with and without insulin and incubated with exogenous substrate poly(Glu 4: Tyr 1). IRS-1 expression was studied in the whole lysate via Western blotting using specific antibodies. Data were analyzed using Student's t-test. RESULTS: The phosphorylation of the exogenous substrate poly(Glu 4: Tyr 1) in myometrium (1.566+/-0.177) and in leiomyoma (1.98+/-0.612) were similar (P=0.774). The IRS-1 levels in myometrium (0.190+/-0.022) and in leiomyoma (0.226+/-0.022) were not different (P=0.184). CONCLUSIONS: There was no difference in IR tyrosine kinase activity (phosphorylation of exogenous substrate) and IRS-1 expression between normal myometrium and leiomyomata. Other steps in the insulin signaling cascade require further study to investigate the role of insulin receptor in leiomyomata.
Assuntos
Leiomioma/metabolismo , Miométrio/metabolismo , Fosfoproteínas/metabolismo , Proteínas Tirosina Quinases/metabolismo , Neoplasias Uterinas/metabolismo , Animais , Feminino , Humanos , Técnicas In Vitro , Insulina/farmacologia , Proteínas Substratos do Receptor de Insulina , Músculo Esquelético/metabolismo , Miométrio/efeitos dos fármacos , Fosforilação , Ratos , Ratos Wistar , Valores de ReferênciaRESUMO
We investigated the transport of (14)C-methylaminoisobutyric acid ((14)C-MeAIB) and (14)C-alanine oxidation in hepatopancreas and jaw muscle of Chasmagnathus granulata submitted to 24, 72, and 144 h of hypo- or hyperosmotic stress. While (14)C-MeAIB uptake increased in jaw muscle and hepatopancreas from crabs submitted to hyperosmotic stress, it did not change in tissues from animals submitted to hypo-osmotic stress. Incubation of jaw muscle and hepatopancreas from control groups with 1 mM ouabain did not decrease (14)C-MeAIB uptake. However, ouabain prevented (14)C-MeAIB uptake in hepatopancreas at 24 h of hyperosmotic stress. In contrast, in jaw muscle from crabs submitted to the same conditions, (14)C-MeAIB uptake was not prevented by ouabain in the incubation medium. Jaw muscle from the control group produced four times more (14)CO(2) from (14)C-alanine than the hepatopancreas. During hypo-osmotic stress, amino acid oxidation does not seem to be one of the pathways implicated in the decrease of the amino acid pools in hepatopancreas and jaw muscle. In contrast, during hyperosmotic stress the reduction in (14)C-alanine oxidation appears to be one of the mechanisms involved in the increase of the amino acid pool in the hepatopancreas.
Assuntos
Aminoácidos/química , Aminoácidos/metabolismo , Alanina/química , Alanina/metabolismo , Animais , Braquiúros/química , Braquiúros/metabolismo , Dióxido de Carbono/metabolismo , Isótopos de Carbono , Hepatopâncreas/química , Hepatopâncreas/metabolismo , Arcada Osseodentária/química , Arcada Osseodentária/metabolismo , Músculos/química , Músculos/metabolismo , Pressão Osmótica , Oxirredução , Fatores de Tempo , beta-Alanina/análogos & derivados , beta-Alanina/química , beta-Alanina/metabolismoRESUMO
We investigated the effects of anoxia (8 h) and different periods of reoxygenation (20 and 40 min) on the oxidative balance in anterior and posterior gills of the crab Chasmagnathus granulata. Enzyme activity of catalase and GST was increased in the gills of the animals submitted to anoxia, and SOD activity was decreased. These enzymes returned approximately to control levels during the anoxia recovery time. These results demonstrated enzyme activities change with variations in environmental oxygen levels. The posterior gills showed a higher antioxidant enzyme activity than anterior gills. In the gills, there were no changes in the non-enzymatic antioxidant system (TRAP) during anoxia. On the other hand, during anoxia recovery, an increase of TRAP in both gills was observed. Anoxia does not change lipid peroxidation (TBARS) in the gills. During anoxia recuperation, an increase in levels of TBARS was observed. Thus the results demonstrate that C. granulata has a similar strategy of preparation for oxidative stress as observed in other intertidal species, enabling the crabs to survive in an environment with extreme variations in physical and chemical characteristics, such as salt marshes.
Assuntos
Antioxidantes/metabolismo , Braquiúros/metabolismo , Brânquias/enzimologia , Hipóxia/metabolismo , Estresse Oxidativo , Oxigênio/metabolismo , Animais , Catalase/metabolismo , Exposição Ambiental , Glutationa Transferase/metabolismo , Peroxidação de Lipídeos , Masculino , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase/metabolismo , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismoRESUMO
The effects of anoxic exposure and the post-anoxia aerobic recovery period on carbohydrate metabolism in the central nervous system (CNS) of the land snail Megalobulimus oblongus, an anoxia-tolerant land gastropod, were studied. The snails were exposed to anoxia for periods of 1.5, 3, 6, 12, 18, or 24 hr. In order to study the post-anoxia recovery phase, snails exposed to a 3-hr period of anoxia were returned to aerobic conditions for 1.5, 3, 6, or 15 hr. Glycogen and glucose concentrations in the CNS, hemolymph glucose concentration, and glycogen phosphorylase (active form, GPa) activity in the CNS were analyzed. Anoxia does not significantly affect the concentration of CNS glucose but induces hyperglycemia and a reduction of CNS GPa activity. The glycogen concentration was decreased at 12 hr of anoxia; however, by 18 and 24 hr in anoxia, the glycogen content was not significantly different from basal control values. During the post-anoxia period, the reduction in GPa activity and the increased hemolymph glucose concentration induced by anoxia returned to control values. These results suggest that the CNS of M. oblongus may use hemolymph glucose to fulfill the metabolic demands during anoxia. However, the hypothesis of tissue metabolic arrest cannot be excluded.
Assuntos
Anaerobiose/fisiologia , Glicemia/metabolismo , Metabolismo dos Carboidratos , Sistema Nervoso Central/metabolismo , Hemolinfa/metabolismo , Caramujos/metabolismo , Adaptação Fisiológica , Análise de Variância , Animais , Sistema Nervoso Central/fisiopatologia , Glicogênio/metabolismo , Recuperação de Função FisiológicaRESUMO
The present study assessed the effect of different fasting times on the in vitro gluconeogenic capacity of Chasmagnathus granulata crabs previously adapted to a high-protein (HP) or carbohydrate-rich (HC) diet using the incorporation of [U-(14)C]l-lactate or [U-(14)C]l-alanine into glucose. We also recorded haemolymphatic glucose and hepatopancreatic glycogen levels. In the HP group, on the third day of fasting there were decreases in the synthesis of glucose from (14)C-alanine and in haemolymph glucose. After 15 days of fasting, haemolymph glucose and hepatopancreatic glycogen levels were maintained by an increase in the conversion of (14)C-alanine into glucose. However, after 21 days of fasting the gluconeogenic capacity was decreased and hepatopancreas glycogen concentration was reduced. In the HC group, hepatopancreatic glycogen was the energy source during the first 6 days of fasting. Gluconeogenesis from (14)C-lactate decreased after 6 days of fasting, remaining low until 21 days of fasting. The conversion of (14)C-alanine into glucose was increased after 15 days fasting and hepatopancreatic glycogen was raised in relation to that present after a 6-day fasting. In both dietary groups the stabilization in the levels of haemolymph glucose after 21 days fasting may result from a reduction in metabolic rate during restricted feeding.
Assuntos
Braquiúros/metabolismo , Jejum , Gluconeogênese , Glicogênio/metabolismo , Hepatopâncreas/metabolismo , Alanina/metabolismo , Animais , Metabolismo dos Carboidratos , Jejum/fisiologia , Gluconeogênese/fisiologia , Glucose/análise , Hemolinfa/química , Ácido Láctico/metabolismo , Masculino , Proteínas/metabolismoRESUMO
Chasmagnathus granulata phosphoenolpyruvate carboxykinase (PEPCK) cDNA from jaw muscle was cloned and sequenced, showing a specific domain to bind phosphoenolpyruvate in addition to the kinase-1 and kinase-2 motifs to bind guanosine triphosphate (GTP) and Mg(2+), respectively, specific for all PEPCKs. In the kinase-1 motifs the GK was changed to RK. The first 19 amino acids of the putative enzyme contain hydrophobic amino acids and hydroxylated residues specific to a mitochondrial type signal. The PEPCK is expressed in hepatopancreas, muscles, nervous system, heart, and gills. Hyperosmotic stress for 24 h increased the PEPCK mRNA level, gluconeogenic and PEPCK activities in muscle.
Assuntos
Gluconeogênese , Músculo Esquelético/metabolismo , Pressão Osmótica , Fosfoenolpiruvato Carboxiquinase (GTP)/genética , Sequência de Aminoácidos , Animais , Sítios de Ligação , Braquiúros , Indução Enzimática , Masculino , Dados de Sequência Molecular , Fosfoenolpiruvato Carboxiquinase (GTP)/biossíntese , RNA Mensageiro/análise , RNA Mensageiro/biossíntese , Alinhamento de Sequência , Análise de Sequência de DNA , Distribuição TecidualRESUMO
C. granulata is a semiterrestrial crab that lives in the mesolittoral and the supralittoral zones of estuaries and faces hypoxia and anoxia when exposed to atmospheric air. The carbohydrate or protein content of the diets administered to the crabs induced different metabolic adjustments during anoxia and post-anoxia recovery period. During the first hour in anoxia a marked increase in L-lactate concentration in hemolymph was induced, followed by a reduction in its levels accompanied by two peaks in hepatopancreas gluconeogenic capacity. Anoxia exposure did not induce a reduction in the hepatopancreas phosphoenolpyruvate carboxykinase activity in either dietary group. Our results suggest that in anaerobiosis this crab uses the conversion of lactate to glucose in hepatopancreas to maintain the acid-base balance and the glucose supply. In post-anoxia recovery, the fate of L-lactate is the hepatopancreas gluconeogenesis in high protein maintained crabs. On the other hand, in the crabs maintained on carbohydrate-rich diet the L-lactate levels decreased gradually in the hemolymph during the post-anoxia recovery; however, the hepatopancreas gluconeogenesis did not increase. In both dietary groups, an increase in the gluconeogenic capacity of hepatopancreas occurred at 30 h of post-anoxia recovery.
Assuntos
Braquiúros/fisiologia , Gluconeogênese/fisiologia , Hepatopâncreas/fisiologia , Anaerobiose/fisiologia , Análise de Variância , Animais , Brasil , Dieta , Carboidratos da Dieta , Proteínas Alimentares , Hepatopâncreas/metabolismo , Ácido Láctico/sangue , Masculino , Fosfoenolpiruvato Carboxiquinase (ATP)/metabolismo , Fatores de TempoRESUMO
The aim of the present study was to examine the effects of insulin on glucose uptake and glycogen synthesis in crab Chasmagnathus granulata gills. We observed an increased glucose uptake and incorporation of d-[(14)C]glucose into glycogen when posterior C. granulata gills were incubated in the presence of insulin; however, this was not observed in anterior gills, despite the presence of similar insulin receptors. In posterior gills, basal glucose uptake in the summer was significantly higher than in the winter. Moreover, in the summer, the insulin dose required to stimulate glucose uptake was twice as high as in the winter. However, there was no significant difference in terms of basal glycogen synthesis in summer and winter. In crustaceans, the endogenous insulin/IGFI substance might be involved in the rapid restoration of glycogen levels in the gills, increasing glucose uptake and glycogen synthesis. Bovine insulin seems to have a stimulatory effect on glycogen metabolism only in posterior gills.