RESUMO
Although UHT heat treatment is being optimized to improve the stability and functional properties of dairy products, its metabolic effects remain scarcely known. As such, we studied the effect of the type of UHT process on lipid metabolism, intestinal barrier, and inflammation in mice. Nine-week-old male C57Bl/6J mice were fed a diet composed of nonlipidic powder mixed with different UHT dairy creams (final: 13% milkfat) for 1 or 4 wk. All creams contained 0.02% of thickener (carrageenan) and were treated via either (1) classical indirect heating process (Th), (2) indirect process at higher temperature (Th+), or (3) direct process by steam injection (ThD). Plasma, epididymal adipose tissue (EAT), and intestine were analyzed. Multivariate principal component analyses were used to identify differential effects of processes. Th+ differed by a globally higher liver damage score compared with that of the other creams. After 4 wk, the duodenal expression of lipid absorption genes fatty acid binding protein 4 (Fatp4) and microsomal triglycerides transfer protein (Mttp) was lower in the Th+ versus Th group. Expression in the colon of tight junction protein zonula occludens 1 (Zo1) and of some endoplasmic reticulum stress markers was lower in both Th+ and ThD versus the Th group. In EAT, ThD had lower gene expression of several inflammatory markers after 4 wk. Some differential effects may be related to heat-induced physicochemical changes of creams. The type of cream UHT process differentially affected metabolic parameters in mice after a 4-wk fat-rich diet, partly due to cream structure. Altogether, direct steam injection process induced the lowest early markers of high-fat-induced metabolic inflammation in EAT.
Assuntos
Tecido Adiposo/imunologia , Laticínios/efeitos adversos , Gorduras/efeitos adversos , Temperatura Alta/efeitos adversos , Leite/química , Tecido Adiposo/metabolismo , Animais , Bovinos , Laticínios/análise , Epididimo/imunologia , Gorduras/química , Gorduras/metabolismo , Intestinos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteína da Zônula de Oclusão-1/imunologiaRESUMO
Additives stabilize or improve the organoleptic or functional properties (or both) of many dairy products including whipping cream. Their influence on the metabolic effect of dairy cream is scarcely known. We tested the hypothesis that added emulsifier (lactic acid esters of mono- and diglycerides; MAG/DAG), thickener (carrageenan, CGN), or both, could modify the metabolic effect, notably in the intestine and liver. Nine-week-old male C57Bl/6J mice were fed UHT cream (indirect treatment) mixed with nonlipidic powder (final: 13% milkfat) for 1 or 4 wk. We compared creams (1) without additive (Ctl), (2) with thickener (Th), 0.02% of κ-CGN, and (3) with both thickener and emulsifier, 0.1% of MAG/DAG esters (Th/Em). We analyzed plasma parameters, intestine, and liver. Fasting glycemia, insulinemia, triglyceridemia, nonesterified fatty acids, body weight gain, and liver weight did not differ among groups. After 1 wk, Th/Em had higher expression in the duodenum of some of the genes involved in (1) intestinal lipid absorption and (2) tight junction proteins versus Ctl and Th. After 4 wk, mucus cell number in the small intestine was higher in Th/Em versus Ctl and Th. Genes involved in endoplasmic reticulum (ER) stress in the duodenum were more expressed in Th/Em after 1 wk. After 4 wk, in the colon, a higher expression of ER stress genes was observed for Th versus Th/Em and Ctl. Liver damage score was not altered by additives. Adding both CGN (0.02%) and MAG/DAG esters (0.1%) in dairy cream did not result in deleterious outcomes in mice after 4 wk regarding lipid metabolism, intestinal permeability, and liver disorders. The longer term effect of intestinal ER stress modulation deserves further investigation.
Assuntos
Laticínios/análise , Emulsificantes/análise , Estresse do Retículo Endoplasmático , Aditivos Alimentares/análise , Intestino Delgado/metabolismo , Metabolismo dos Lipídeos , Animais , Bovinos , Duodeno/metabolismo , Emulsificantes/metabolismo , Ácidos Graxos não Esterificados/sangue , Aditivos Alimentares/metabolismo , Fígado/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Leite/químicaRESUMO
Silicon nanoparticles (SiNPs) prepared by mechanical grinding of luminescent porous silicon were coated with a biopolymer (dextran) and investigated as a potential theranostic agent for bioimaging and sonodynamic therapy. Transmission electron microscopy, photoluminescence and Raman scattering measurements of dextran-coated SiNPs gave evidence of their enhanced stability in water. In vitro experiments confirmed the lower cytotoxicity of the dextran-coated NPs in comparison with uncoated ones, especially for high concentrations of about 2 mg ml-1. Efficient uptake of the NPs by cancer cells was found using bioimaging in the optical transmittance and photoluminescence modes. Treatment of the cells with uptaken SiNPs by therapeutic ultrasound for 5-20 min resulted in a strong decrease in the number of living cells, while the total number of cells remained nearly unchanged. The obtained data indicate a 'mild' effect of the combined action of ultrasonic irradiation and SiNPs on cancer cells. The observed results reveal new opportunities for controlling the photoluminescent and sonosensitizing properties of silicon-based NPs for applications in the diagnostics and mild therapy of cancer.
Assuntos
Materiais Revestidos Biocompatíveis/química , Nanopartículas/toxicidade , Neoplasias/patologia , Silício/toxicidade , Nanomedicina Teranóstica/métodos , Ondas Ultrassônicas , Células 3T3-L1 , Animais , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Dextranos/química , Cães , Humanos , Luminescência , Camundongos , Nanopartículas/química , Nanopartículas/ultraestrutura , Análise Espectral Raman , Fatores de TempoRESUMO
Adipose tissue and adipocytes play a central role in the pathogenesis of metabolic diseases related to obesity. Size of fat cells depends on the balance of synthesis and mobilization of lipids and can undergo important variations throughout the life of the organism. These variations usually occur when storing and releasing lipids according to energy demand. In particular when confronted to severe food restriction, adipocyte releases its lipid content via a process called lipolysis. We propose a mathematical model that combines cell diameter distribution and lipolytic response to show that lipid release is a surface (radius squared) limited mechanism. Since this size-dependent rate affects the cell׳s shrinkage speed, we are able to predict the cell size distribution evolution when lipolysis is the only factor at work: such as during an important food restriction. Performing recurrent surgical biopsies on rats, we measured the evolution of adipose cell size distribution for the same individual throughout the duration of the food restriction protocol. We show that our microscopic model of size dependent lipid release can predict macroscopic size distribution evolution.
Assuntos
Tecido Adiposo/citologia , Privação de Alimentos/fisiologia , Modelos Biológicos , Animais , Contagem de Células , Tamanho Celular , Ingestão de Energia , Lipólise , Tamanho do Órgão , Ratos WistarRESUMO
OBJECTIVE: During septic shock, vasopressors are a cornerstone of therapy. In septic shock, very high doses of vasopressors sometimes have to be used due to vascular desensitization, the mechanisms of which are poorly understood. This study assesses whether α-2 agonists increase pressor responsiveness following lipopolysaccharide administration. DESIGN: Parallel groups of animals (n = 7 per group) subjected to pharmacologic interventions. SETTING: Physiology laboratory. SUBJECTS: Rats. INTERVENTIONS: In anesthetized rats, the pressor responses to increasing doses of norepinephrine (norepinephrine-systolic pressure curve) were assessed during a baseline period, after injection of saline or lipopolysaccharide, and after subsequent injection of saline, dexmedetomidine (100 µg/kg IV), or clonidine (200 µg/kg IV). MEASUREMENTS AND MAIN RESULTS: Differences in the slopes of the norepinephrine-pressure curves were assessed across drug treatments and intervals. The pressor dose of norepinephrine necessary to increase systolic pressure by 33 and 100 mm Hg (pressor dose 33 and pressor dose 100) was determined. Pressor responsiveness to norepinephrine decreased slightly over time in the saline-saline group (saline 1 or 2 vs baseline: mean decrease of the slope, 2 mm Hg/µg/kg norepinephrine; p < 0.05), whereas there was a large decrease after lipopolysaccharide (lipopolysaccharide vs baseline: mean decrease of the slope, 7.2; p < 0.001). Clonidine alone had no effect, but when administered following lipopolysaccharide, it caused a striking increase in pressor responsiveness (mean slope after lipopolysaccharide, 10.7 [95% CI, 9.9-11.6]; after clonidine, 17.5 [95% CI, 16.7-18.4]). Similarly, dexmedetomidine administered after lipopolysaccharide caused a large increase in pressor responsiveness above lipopolysaccharide values. Accordingly the pressor dose 33 and pressor dose 100 values were lowered following lipopolysaccharide and restored by α-2 agonists. CONCLUSIONS: The pressor response to norepinephrine was reduced following lipopolysaccharide and increased to baseline levels following α-2 agonists.
Assuntos
Agonistas de Receptores Adrenérgicos alfa 2/uso terapêutico , Pressão Arterial/efeitos dos fármacos , Clonidina/uso terapêutico , Dexmedetomidina/uso terapêutico , Hipotensão/tratamento farmacológico , Choque Séptico/fisiopatologia , Animais , Hipotensão/etiologia , Hipotensão/fisiopatologia , Lipopolissacarídeos , Masculino , Norepinefrina/uso terapêutico , Projetos Piloto , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley , Choque Séptico/induzido quimicamente , Choque Séptico/complicações , Vasoconstritores/uso terapêuticoRESUMO
Adipocytes are cells whose task is to store excess energy as lipid droplets in their cytoplasm. Adipocytes can adapt their size according to the lipid amount to be stored. Adipocyte size variation can reach one order of magnitude inside the same organism which is unique among cells. A striking feature in adipocytes size distribution is the lack of characteristic size since typical size distributions are bimodal. Since energy can be stored and retrieved and adipocytes are responsible for these lipid fluxes, we propose a simple model of size-dependent lipid fluxes that is able to predict typical adipocytes size distribution.
Assuntos
Adipócitos/citologia , Adipócitos/metabolismo , Tamanho Celular , Metabolismo dos Lipídeos/fisiologia , Modelos Biológicos , Animais , Humanos , RatosRESUMO
Covalent grafting of amino groups onto the carboxylic acid functionalities, naturally covering the surface of fluorescent nanoparticles produced from silicon carbide (SiC NPs), allowed tuning of their surface charge from negative to highly positive. Incubating 3T3-L1 fibroblast cells with differently charged SiC NPs demonstrates the crucial role of the charge in cell fluorescent targeting. Negatively charged SiC NPs concentrate inside the cell nuclei. Close to neutrally charged SiC NPs are present in both cytoplasm and nuclei while positively charged SiC NPs are present only in the cytoplasm and are not able to move inside the nuclei. This effect opens the door for the use of SiC NPs for easy and fast visualization of long-lasting biological processes taking place in the cell cytosol or nucleus as well as providing a new long-term cell imaging tool. Moreover, here we have shown that the interaction between charged NPs and nuclear pore complex plays an essential role in their penetration into the nuclei.
Assuntos
Corantes Fluorescentes/análise , Nanopartículas/análise , Poro Nuclear/metabolismo , Proteínas/metabolismo , Células 3T3-L1 , Aminas/química , Animais , Compostos Inorgânicos de Carbono/química , Ácidos Carboxílicos/química , Núcleo Celular/química , Citoplasma/química , Corantes Fluorescentes/química , Camundongos , Nanopartículas/química , Poro Nuclear/química , Tamanho da Partícula , Transporte Proteico , Compostos de Silício/química , Espectrometria de Fluorescência , Espectroscopia de Infravermelho com Transformada de Fourier , Coloração e Rotulagem , Eletricidade Estática , Propriedades de SuperfícieRESUMO
BACKGROUND: Adipose tissue engineering has been hampered by the inability to culture mature adipocytes. Adipose-derived stem cell (ASC) culture opens the way for the preparation of human 3-D hypodermis in large quantities. These models play a role in obesity-related active molecules and slimming agent screening. Moreover, they contribute to a better understanding of the mechanisms underpinning obesity. MATERIALS AND METHODS: Freshly extracted ASC from fat tissue were characterized by flow cytometry for CD73, CD90, CD105, HLA-ABC, CD14 and CD45 markers and by Western blot for pref-1. Their differentiation in mature adipocytes was followed by lipid and adiponectin secretion or by oil red O staining and radioimmunoassay. Neosynthesized extracellular matrix (ECM) of 3-D hypodermis was investigated by immunohistochemistry (collagen type I, V and VI) and transmission electron microscopy. RESULTS: Our results demonstrate that the culture of preadipocytes in proliferation medium for 15 days followed by 16 days of culture in differentiation medium allowed production of the thickest single-layer hypodermis in which preadipocytes and mature adipocytes coexist and synthesize adiponectin and ECM components. Functionality of our 3-D single-layer hypodermis was demonstrated both by a 3.5-fold glycerol production after its stimulation with norepinephrine (adrenergic agonist) and by its slimming after caffeine treatment versus the nontreated 3-D hypodermis. CONCLUSION: This economic 3-D model, easy to prepare and giving reproducible results after the treatment of actives, is useful for pharmacotoxicological trials as an alternative to animal experimentation.
Assuntos
Tecido Adiposo/citologia , Tela Subcutânea , Engenharia Tecidual/métodos , Adiponectina/metabolismo , Antígenos CD/metabolismo , Cafeína/farmacologia , Diferenciação Celular , Células Cultivadas , Humanos , Microscopia Eletrônica de Transmissão , Pessoa de Meia-Idade , Fenótipo , Células-Tronco/citologia , Tela Subcutânea/efeitos dos fármacos , Tela Subcutânea/metabolismo , Tela Subcutânea/ultraestruturaRESUMO
OBJECTIVE: We previously reported that the flavonoid cirsimarin exerts in vitro a strong lipolytic activity on isolated adipocytes. This study was therefore designed to evaluate in vivo the effects of cirsimarin on white adipose tissue (WAT) accretion in mice. METHODS: Male CD1 mice were injected daily with either vehicle (intraperitoneal (i.p.)) or cirsimarin (25 or 50 mg kg(-1) per day, i.p.) for 18 days. Mice were killed and fat pads weighted. Epididymal fat pads were used for cellularity measurement. Effects of cirsimarin treatment on lipolysis and lipogenesis in WAT were assessed. RESULTS: Mice treated with 25 or 50 mg kg(-1) per day cirsimarin showed a decrease in retroperitoneal (-29 and -37% respectively, P<0.005) and epididymal (-25 and -28% respectively, P<0.005) fat pad weights compared with controls. This effect was restricted to intra-abdominal WAT as no difference was noticed for subcutaneous inguinal WAT. The decrease in intra-abdominal WAT accretion was due to a decrease in adipose cell diameter (-5 and -8% for 25 and 50 mg kg(-1) per day cirsimarin, respectively) resulting in a 14 and 35% decrease in adipose cell volume while no change was noticed in total adipocyte number. Direct injection of cirsimarin (50 mg kg(-1)) to rats did not trigger lipolysis. In contrast, cirsimarin showed in vivo as well as in vitro a strong antilipogenic activity, which may be the critical aspect of its effects on fat accretion in mice. The inhibitory concentration 50% of cirsimarin on lipogenic activity in isolated adipocytes was found to be 1.28±0.04 µM. Cirsimarin given orally reduced intra-abdominal fat accretion in mice. CONCLUSION: Cirsimarin exerts potent antilipogenic effect and decreases adipose tissue deposition in mice. Cirsimarin could therefore be a potential candidate for the treatment of obesity.
Assuntos
Adipócitos/efeitos dos fármacos , Peso Corporal/efeitos dos fármacos , Flavonas/farmacologia , Glicosídeos/farmacologia , Gordura Intra-Abdominal/efeitos dos fármacos , Lipogênese/efeitos dos fármacos , Adipócitos/metabolismo , Animais , Peso Corporal/fisiologia , Contagem de Células , Tamanho Celular/efeitos dos fármacos , Flavonas/administração & dosagem , Glicosídeos/administração & dosagem , Injeções Intraperitoneais , Gordura Intra-Abdominal/metabolismo , Lipogênese/fisiologia , Masculino , Camundongos , RatosRESUMO
During severe septic shock and/or severe acute respiratory distress syndrome (ARDS) patients present with a limited cardio-ventilatory reserve (low cardiac output and blood pressure, low mixed venous saturation, increased lactate, low PaO2/FiO2 ratio, etc.), especially when elderly patients or co-morbidities are considered. Rescue therapies (low dose steroids, adding vasopressin to noradrenaline, proning, almitrine, NO, extracorporeal membrane oxygenation, etc.) are complex. Fever, above 38.5-39.5°C, increases both the ventilatory (high respiratory drive: large tidal volume, high respiratory rate) and the metabolic (increased O2 consumption) demands, further limiting the cardio-ventilatory reserve. Some data (case reports, uncontrolled trial, small randomized prospective trials) suggest that control of elevated body temperature ("fever control") leading to normothermia (35.5-37°C) will lower both the ventilatory and metabolic demands: fever control should simplify critical care management when limited cardio-ventilatory reserve is at stake. Usually fever control is generated by a combination of general anesthesia ("analgo-sedation", light total intravenous anesthesia), antipyretics and cooling. However general anesthesia suppresses spontaneous ventilation, making the management more complex. At variance, alpha-2 agonists (clonidine, dexmedetomidine) administered immediately following tracheal intubation and controlled mandatory ventilation, with prior optimization of volemia and atrio-ventricular conduction, will reduce metabolic demand and facilitate normothermia. Furthermore, after a rigorous control of systemic acidosis, alpha-2 agonists will allow for accelerated emergence without delirium, early spontaneous ventilation, improved cardiac output and micro-circulation, lowered vasopressor requirements and inflammation. Rigorous prospective randomized trials are needed in subsets of patients with a high fever and spiraling toward refractory septic shock and/or presenting with severe ARDS.
RESUMO
Oxidative stress and low grade chronic inflammation are increased in accumulating fat. Our objective was to test whether 4-hydroxynonenal (4-HNE), an end-product of lipid peroxidation, affects cyclooxygenases in 3T3-L1 adipose cells. 4-HNE increased COX-2 mRNA and protein expression and p38MAP-kinase phosphorylation in a dose-dependent manner. Pretreatment of 3T3-L1 cells by a selective inhibitor of p38MAPK (PD 169316) abolished 4-HNE and glucose oxidase induced COX-2 expression. Our results show that oxidative stress induces COX-2 expression through the production of 4-HNE which activates p38MAPKinase, suggesting that 4-HNE links oxidative stress and chronic inflammation through the activation of cyclooxygenase.
Assuntos
Adipócitos/enzimologia , Aldeídos/farmacologia , Ciclo-Oxigenase 2/genética , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Estresse Oxidativo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Células 3T3 , Adipócitos/efeitos dos fármacos , Animais , Inibidores de Cisteína Proteinase/farmacologia , Dinoprosta/biossíntese , Dinoprostona/biossíntese , Ativação Enzimática , Peroxidação de Lipídeos , Camundongos , Prostaglandina-Endoperóxido Sintases/genética , RNA Mensageiro/genéticaRESUMO
Obesity is an increasing nutritional disorder in developed countries, and oxidative stress has been identified as a key factor in numerous pathologies such as diabetes, inflammation, and atherosclerosis, which are favored by obesity. The objective of the present study was to investigate the effects of oxidative stress in 3T3-L1 adipose cells on two parameters involved in metabolic complications associated with obesity, namely adiponectin secretion and lactate production. Differentiated 3T3-L1 adipose cells were exposed to increasing concentrations of glucose oxidase. 4-Hydroxynonenal (4-HNE), a relevant lipid peroxidation by-product which may affect several metabolic processes in making covalent adducts with various molecules; adiponectin secretion; and lactate production were measured in response to glucose oxidase exposure. Results show an inhibition of adiponectin mRNA expression by glucose oxidase and a significant inverse correlation between 4-HNE formation and adiponectin secretion. Furthermore, 4-HNE alone inhibits adiponectin production by 3T3-L1. On the other hand, glucose oxidase and 4-HNE significantly stimulated lactate production by 3T3-L1 adipocytes. These results demonstrate that adipose cells are highly sensitive to oxidative stress, with subsequent decreased adiponectin secretion and increased lactate production, two events involved in the development of insulin resistance.
Assuntos
Adipócitos/metabolismo , Peptídeos e Proteínas de Sinalização Intercelular/biossíntese , Ácido Láctico/biossíntese , Estresse Oxidativo , Células 3T3-L1 , Adipócitos/efeitos dos fármacos , Adiponectina , Aldeídos/metabolismo , Animais , Glucose Oxidase/farmacologia , Peptídeos e Proteínas de Sinalização Intercelular/genética , Camundongos , RNA Mensageiro/análise , RNA Mensageiro/metabolismoRESUMO
Obesity is linked to adipose tissue hypertrophy (increased adipocyte cell size) and hyperplasia (increased cell number). Comparative analyses of gene datasets allowed us to identify 1426 genes which may represent common adipose phenotype in humans and mice. Among them we identified several adipocyte-specific genes dysregulated in obese adipose tissue, involved in either fatty acid storage (acyl CoA synthase ACSL1, hormone-sensitive lipase LIPE, aquaporin 7 AQP7, perilipin PLIN) or cell adhesion (fibronectin FN1, collagens COL1A1, COL1A3, metalloprotein MMP9, or both (scavenger receptor FAT/CD36). Using real-time analysis of cell surface occupancy on xCELLigence system we developed a new method to study lipid uptake and differentiation of mouse 3T3L1 fibroblasts and human adipose stem cells. Both processes are regulated by insulin and fatty acids such as oleic acid. We showed that fatty acid addition to culture media increased the differentiation rate and was required for full differentiation into unilocular adipocytes. Significant activation of lipogenesis, i.e. lipid accumulation, by either insulin or oleic acid was monitored in times ranging from 1 to 24 h, depending on differentiation state, whereas significant effects on adipogenesis, i.e., surperimposed lipid accumulation and gene transcriptional regulations were measured after 3 to 4 d. Combination of selected times for analysis of lipid contents, cell counts, size fractionations, and gene transcriptional regulations showed that FAT/CD36 specific inhibitor AP5258 significantly increased cell survival of oleic acid-treated mouse and human adipocytes, and partially restored the transcriptional response to oleic acid in the presence of insulin through JNK pathway. Taken together, these data open new perspectives to study the molecular mechanisms commonly dysregulated in mouse and human obesity at the level of lipogenesis linked to hypertrophy and adipogenesis linked to hyperplasia.
RESUMO
Progress over the last 50 years has led to a decline in mortality from ≈70% to ≈20% in the best series of patients with septic shock. Nevertheless, refractory septic shock still carries a mortality close to 100%. In the best series, the mortality appears related to multiple organ failure linked to comorbidities and/or an intense inflammatory response: shortening the period that the subject is exposed to circulatory instability may further lower mortality. Treatment aims at reestablishing circulation within a "central" compartment (i.e., brain, heart, and lung) but fails to reestablish a disorganized microcirculation or an adequate response to noradrenaline, the most widely used vasopressor. Indeed, steroids, nitric oxide synthase inhibitors, or donors have not achieved overwhelming acceptance in the setting of septic shock. Counterintuitively, α 2-adrenoceptor agonists were shown to reduce noradrenaline requirements in two cases of human septic shock. This has been replicated in rat and sheep models of sepsis. In addition, some data show that α 2-adrenoceptor agonists lead to an improvement in the microcirculation. Evidence-based documentation of the effects of alpha-2 agonists is needed in the setting of human septic shock.
Assuntos
Inflamação/tratamento farmacológico , Insuficiência de Múltiplos Órgãos/tratamento farmacológico , Norepinefrina/metabolismo , Choque Séptico/tratamento farmacológico , Agonistas de Receptores Adrenérgicos alfa 2/metabolismo , Agonistas de Receptores Adrenérgicos alfa 2/uso terapêutico , Animais , Clonidina/uso terapêutico , Dexmedetomidina/uso terapêutico , Humanos , Inflamação/mortalidade , Inflamação/patologia , Insuficiência de Múltiplos Órgãos/mortalidade , Insuficiência de Múltiplos Órgãos/patologia , Ratos , Ovinos , Choque Séptico/mortalidade , Choque Séptico/patologiaRESUMO
White adipose tissue is known to contain the components of the renin-angiotensin system, which gives rise to angiotensin II from angiotensinogen (AGT). Recent evidence obtained in vitro and ex vivo is in favor of angiotensin II acting as a trophic factor of adipose tissue development. To determine whether AGT plays a role in vivo in this process, comparative studies were performed in AGT-deficient (agt(-/-)) mice and control wild-type mice. The results showed that agt(-/-) mice gain less weight than wild-type mice in response to a chow or high fat diet. Adipose tissue mass from weaning to adulthood appeared altered rather specifically, as both the size and the weight of other organs were almost unchanged. Food intake was similar for both genotypes, suggesting a decreased metabolic efficiency in agt(-/-) mice. Consistent with this hypothesis, cellularity measurement indicated hypotrophy of adipocytes in agt(-/-) mice with a parallel decrease in the fatty acid synthase activity. Moreover, AGT-deficient mice exhibited a significantly increased locomotor activity, whereas metabolic rate and mRNA levels of uncoupling proteins remained similar in both genotypes. Thus, AGT appears to be involved in the regulation of fat mass through a combination of decreased lipogenesis and increased locomotor activity that may be centrally mediated.
Assuntos
Tecido Adiposo/crescimento & desenvolvimento , Angiotensinogênio/deficiência , Dieta , Atividade Motora/fisiologia , Aumento de Peso , Tecido Adiposo/patologia , Tecido Adiposo Marrom/crescimento & desenvolvimento , Tecido Adiposo Marrom/patologia , Angiotensinogênio/genética , Animais , Camundongos , Camundongos Endogâmicos ICR , Camundongos Knockout/genética , Valores de Referência , TermogêneseRESUMO
The role of the sympathetic nervous system in the regulation by insulin of the level of uncoupling protein in brown adipose tissue has been examined. The amount of uncoupling protein was substantially reduced in streptozotocin-diabetic rats, while insulin replacement to diabetic animals induced a partial restoration. Unilateral denervation of the interscapular brown fat pads also lowered the amount of uncoupling protein, and in diabetic animals inhibited the stimulation of the level of the protein by insulin replacement. Maintenance of normal uncoupling protein levels requires both insulin and the sympathetic system; regulation of the protein by insulin involves sympathetic mediation.
Assuntos
Tecido Adiposo Marrom/metabolismo , Proteínas de Transporte , Insulina/farmacologia , Proteínas de Membrana/metabolismo , Sistema Nervoso Simpático/fisiologia , Tecido Adiposo Marrom/efeitos dos fármacos , Animais , Peso Corporal/efeitos dos fármacos , Denervação , Diabetes Mellitus Experimental/metabolismo , Canais Iônicos , Proteínas Mitocondriais , Ratos , Proteína Desacopladora 1RESUMO
The control of uncoupling protein-1, -2 and -3 (UCP-1, UCP-2, UCP-3) mRNA levels by sympathetic innervation in rats was investigated by specific and sensitive RT-PCR assays. In rats reared at thermoneutrality (25 degrees C), unilateral surgical sympathetic denervation of interscapular brown adipose tissue (BAT) markedly reduced the UCP-1 mRNA level (-38%) as compared with the contralateral innervated BAT pad, but was without significant effect on UCP-2 and -3 mRNA levels. Cold exposure (7 days, 4 degrees C) markedly increased UCP-1 (+180%), UCP-2 (+115%) and UCP-3 (+195%) mRNA levels in interscapular BAT. Unilateral sympathetic denervation prevented the cold-induced rise in BAT UCP-1 and UCP-2 mRNAs, but not that in BAT UCP-3 mRNA. Results were confirmed by Northern blot analysis. These data indicate a differential endocrine control of UCP-1, UCP-2 and UCP-3 gene expression in rat BAT both at thermoneutrality and during prolonged cold exposure.
Assuntos
Tecido Adiposo Marrom/metabolismo , Proteínas de Transporte/genética , Regulação da Expressão Gênica/genética , Proteínas de Membrana/genética , Proteínas de Membrana Transportadoras , Proteínas Mitocondriais , Proteínas/genética , Sistema Nervoso Simpático/fisiologia , Análise de Variância , Animais , Temperatura Baixa , Canais Iônicos , Masculino , RNA Mensageiro/genética , Ratos , Ratos Wistar , Simpatectomia , Temperatura , Proteína Desacopladora 1 , Proteína Desacopladora 2 , Proteína Desacopladora 3RESUMO
The objective of the present study was to test in vivo the metabolic effects of 3,5-di-iodothyronine (3,5-T2) in unanesthetized and unrestrained male Sprague-Dawley rats. Amino acid and lipid metabolisms were investigated by breath tests using as tracers the 13C-carboxyl-labeled molecules of leucine, alpha-ketoisocaproic acid (KIC) and octanoic acid, in four different groups of rats: hypothyroid animals (receiving propylthiouracil (PTU) and iopanoic acid), hypothyroid animals treated with either a daily i.p. injection of 3,5-T2 (25 micrograms/100 g body weight), or tri-iodothyronine (T3) (1 microgram/100 g body weight), and control euthyroid animals receiving equivalent volumes of the vehicle solutions. Energy expenditure was measured by continuous monitoring of O2 consumption and CO2 production in these different groups. Daily energy expenditure was decreased by 30% in PTU-treated rats. The chronic treatments with 3,5-T2 and T3 restored daily energy expenditure to the control level. 13CO2 recovered in breath following the i.v. injection of octanoic acid-[1-13C] was decreased in hypothyroid animals compared with control animals (P < 0.05) and restored to control values by T3 and 3,5-T2 treatments. The 13CO2 recovered in breath after i.v. injection of leucine-[1-13C] was increased in PTU-treated compared with control animals (P < 0.05). Chronic treatment with either 3,5-T2 or T3 restored 13CO2 to control values. Excretion of 13CO2 recovered in breath following the i.v. injection of KIC-[1-13C] was increased in PTU-treated compared with control animals. Chronic treatments with either 3,5-T2 or T3 did not restore KIC decarboxylation. These results suggest that 3,5-T2 exerts metabolic effects on energy expenditure, on both lipid beta-oxidation and leucine metabolism in hypothyroid rats. We conclude that 3,5-T2 is a metabolically active iodothyronine.
Assuntos
Aminoácidos/metabolismo , Di-Iodotironinas/farmacologia , Metabolismo Energético/efeitos dos fármacos , Hipotireoidismo/metabolismo , Metabolismo dos Lipídeos , Animais , Testes Respiratórios , Caproatos/metabolismo , Caprilatos/metabolismo , Di-Iodotironinas/metabolismo , Cetoácidos/metabolismo , Leucina/metabolismo , Masculino , Propiltiouracila , Ratos , Ratos Sprague-Dawley , Tiroxina/sangue , Tri-Iodotironina/sangue , Tri-Iodotironina/farmacologiaRESUMO
Lipolytic responsiveness of subcutaneous and epididymal adipose tissue to norepinephrine (NE) was measured by microdialysis before and during a euglycemic-hyperinsulinemic clamp in male Sprague-Dawley rats (280 +/- 7g, n = 8). Microdialysis probes were perfused with standard Krebs-Ringer buffer without (basal condition [BC]) or with NE 10(-6) mol/L to determine basal and stimulated rates of lipolysis. The dialysate concentration of glycerol was measured (lipolytic index). NE infusion resulted in 3.0- and 4.2-fold increases in glycerol release in abdominal subcutaneous and epididymal adipose tissues, respectively. A euglycemic-hyperinsulinemic clamp at 6 mU/kg.min increased by ninefold the insulinemia (120 +/- 9 U/L). Hyperinsulinemia suppressed basal glycerol release by 57% and 42% in subcutaneous and epididymal adipose depots, respectively (BC + I). Lipolytic responses to NE infusion during a euglycemic-hyperinsulinemic clamp (NE + I) were reduced by 45% and 33% in subcutaneous and epididymal adipose tissues, respectively, as compared with BC. Under BC, the lipolytic response to NE was greater in epididymal than in subcutaneous adipose tissue. Physiological levels of insulin regulated basal lipolysis and counteracted adrenergic stimulation of lipolysis to a similar extent in both superficial (subcutaneous) and intraabdominal (epididymal) adipose tissue. Our findings show that lipolysis is more responsive to NE in epididymal than in subcutaneous adipose tissue. The antilipolytic effects of insulin are similar in both superficial and deep intraabdominal adipose tissues. Furthermore, physiological plasma insulin levels cannot fully antagonize the lipolytic effects of NE.
Assuntos
Insulina/farmacologia , Lipólise/efeitos dos fármacos , Norepinefrina/farmacologia , Tecido Adiposo/irrigação sanguínea , Tecido Adiposo/efeitos dos fármacos , Tecido Adiposo/metabolismo , Animais , Epididimo , Técnica Clamp de Glucose , Glicerol/metabolismo , Mucosa Intestinal/metabolismo , Masculino , Microdiálise , Norepinefrina/administração & dosagem , Ratos , Ratos Sprague-Dawley , Fluxo Sanguíneo Regional , PeleRESUMO
The present study was designed to investigate the hormonal regulation of rat liver glycogenolysis in growth hormone (GH) deficiency. To this end, hepatocytes were isolated from control, GH-deprived (hypophysectomized and treated with triiodothyronine [T3] and corticotropin), and 7-day GH-supplemented fed rats and incubated with glucagon and alpha 1-adrenergic agonist (phenylephrine) to measure the hormonal activation of both glycogen phosphorylase and glucose production from glycogen stores. GH deficiency induces a combined decrease of 50% of the glycogen content, the activity of glucose-6-phosphatase, and the maximal hormone-induced glycogen phosphorylase activity. Daily GH injections restore the levels of both glycogen phosphorylase and glucose-6-phosphatase. These enzymatic inductions occur without normalization of insulinemia. Despite the reduced levels of key enzymes of glycogenolysis, the stimulation of glucose production from glycogen in response to glucagon and phenylephrine is not modified in GH-deprived rats. An increase in the intrinsic activity of one or both of the enzymatic steps is postulated to compensate for the lower levels of enzymes, as indicated by the slopes of the correlation between glucose production and phosphorylase a activity (107 and 216 nmol glucose produced/min/U phosphorylase a [P < .001] in control and GH-deprived rats, respectively). GH replacement enhances maximal phosphorylase activity and brings the correlation toward the control value (slope, 128 nmol glucose produced/min/U phosphorylase a). Our findings demonstrate that glycogenolysis in hepatocytes isolated from GH-deprived rats is normal, despite a reduction of glycogen phosphorylase and glucose-6-phosphatase activities.(ABSTRACT TRUNCATED AT 250 WORDS)