Assuntos
Diabetes Mellitus Tipo 2 , Obesidade , Humanos , Obesidade/epidemiologia , Prevalência , Espanha/epidemiologia , Uso de TabacoRESUMO
Since the discovery of leptin in 1994, the adipose tissue (AT) is not just considered a passive fat storage organ but also an extremely active secretory and endocrine organ that secretes a large variety of hormones, called adipokines, involved in energy metabolism. Adipokines may not only contribute to AT dysfunction and obesity, but also in fat browning, a process that induces a phenotypic switch from energy-storing white adipocytes to thermogenic brown fat-like cells. The fat browning process and, consequently, thermogenesis can also be stimulated by physical exercise. Contracting skeletal muscle is a metabolically active tissue that participates in several endocrine functions through the production of bioactive factors, collectively termed myokines, proposed as the mediators of physical activity-induced health benefits. Myokines affect muscle mass, have profound effects on glucose and lipid metabolism, and promote browning and thermogenesis of white AT in an endocrine and/or paracrine manner. The present review focuses on the role of different myokines and adipokines in the regulation of fat browning, as well as in the potential cross-talk between AT and skeletal muscle, in order to control body weight, energy expenditure and thermogenesis.
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Adipocinas/fisiologia , Tecido Adiposo Marrom/metabolismo , Músculo Esquelético/metabolismo , Obesidade/metabolismo , Tecido Adiposo Marrom/citologia , Animais , Metabolismo Energético , Exercício Físico , Humanos , Músculo Esquelético/citologia , TermogêneseRESUMO
BACKGROUND/OBJECTIVES: Bariatric surgery improves nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH), but the underlying mechanisms remain elusive. We evaluated the potential role of ghrelin isoforms in the amelioration of hepatic inflammation after sleeve gastrectomy and Roux-en-Y gastric bypass (RYGB). SUBJECTS/METHODS: Plasma ghrelin isoforms were measured in male Wistar rats (n = 129) subjected to surgical (sham operation, sleeve gastrectomy, or RYGB) or dietary interventions [fed ad libitum a normal (ND) or a high-fat diet (HFD) or pair-fed diet]. The effect of acylated and desacyl ghrelin on markers of inflammation, mitochondrial dysfunction, and endoplasmic reticulum (ER) stress in primary rat hepatocytes under palmitate-induced lipotoxic conditions was assessed. RESULTS: Plasma desacyl ghrelin was decreased after sleeve gastrectomy and RYGB, whereas the acylated/desacyl ghrelin ratio was augmented. Both surgeries diminished obesity-associated hepatic steatosis, CD68+- and apoptotic cells, proinflammatory JNK activation, and Crp, Tnf, and Il6 transcripts. Moreover, a postsurgical amelioration in the mitochondrial DNA content, oxidative phosphorylation (OXPHOS) complexes I and II, and ER stress markers was observed. Specifically, following bariatric surgery GRP78, spliced XBP-1, ATF4, and CHOP levels were reduced, as were phosphorylated eIF2α. Interestingly, acylated and desacyl ghrelin inhibited steatosis and inflammation of palmitate-treated hepatocytes in parallel to an upregulation of OXPHOS complexes II, III, and V, and a downregulation of ER stress transducers IRE1α, PERK, ATF6, their downstream effectors, ATF4 and CHOP, as well as chaperone GRP78. CONCLUSIONS: Our data suggest that the increased relative acylated ghrelin levels after bariatric surgery might contribute to mitigate obesity-associated hepatic inflammation, mitochondrial dysfunction, and ER stress.
Assuntos
Cirurgia Bariátrica , Estresse do Retículo Endoplasmático/fisiologia , Grelina , Hepatite/metabolismo , Mitocôndrias/metabolismo , Acilação , Animais , Células Cultivadas , Grelina/análogos & derivados , Grelina/sangue , Grelina/química , Grelina/metabolismo , Hepatócitos/metabolismo , Masculino , Mitocôndrias/patologia , Isoformas de Proteínas , Ratos , Ratos WistarRESUMO
OBJETIVO: Determinar el rendimiento diagnóstico del PET/CT en el estudio de nódulo pulmonar (NP) utilizando SUVmax. MÉTODO: Se revisó la base de datos de PET/CT, seleccionando aquellos solicitados para estudio de NP sólido. Se incluyeron sólo aquellos NP confirmados como malignos o benignos. Se excluyó NP subsólidos, masas pulmonares (> 3 cm), y pacientes con metástasis conocidas. Se midió SUVmax de las lesiones, determinando mejores valores de corte para malignidad y benignidad. RESULTADOS: De los 140 NP estudiados, el 60% (84/140) fueron confirmados como malignos y el 40% como benignos (100% y 59,6% de confirmación histológica, respectivamente). Un SUVmax ≤ 1,0 mostró sensibilidad 98,8%, valor predictivo negativo (VPN) 96,2%, y Likelihood ratio negativo (LR -) 0,027. Un SUVmax ≤ 2,5 no fue capaz de asegurar razonablemente benignidad con VPN 69,4%, y LR - 0,295. Valores de SUV > 2,5 y 5,0 se asociaron a malignidad en 83% y 93% de los casos, respectivamente (LR+ 3,333 y 8,889). CONCLUSIÓN: El PET/CT presenta alto rendimiento diagnóstico en estimar la naturaleza de un NP Un valor de SUVmax ≤ 1 es altamente predictivo de benignidad, y un valor de SUVmax ≥ 2,5 de malignidad. Valores entre 1,0 y 2,5 no permiten caracterizar eficientemente los NP.
AIM: To establish the diagnostic accuracy of PET/CT in study of solid lung nodule (LN) using SUVmax index. METHOD: We revised PET/CT data base, selecting those scans asked to evaluate a solid LN. Only confirmed malign o benign LN were included. Subsolid LN, lung masses (> 3 cm), and known or suspected lung metastases were excluded. SUVmax was measured in each LN, and best cutoff for malignant and benign lesion was calculated. RESULTS: Of the whole group of 140 LN, 60% were confirmed as malignant, and 40% as benign (100% and 59,6% of histological confirmation, respectively). SUVmax ≤ 1,0 showed sensibility of 98,8%, negative predictive value (NPV) of 96,2%, and negative likelihood ratio (LR —) of 0,027. SUVmax ≤ 2,5 was not able to guarantee reasonably benign nature of LN, showing NPV of 69,4% and LR - of 0,295. SUVmax > 2,5 and > 5,0 was associated to malign lesion in 83% and 93% of cases, respectively (LR + of 3,333 and 8,889). CONCLUSION: PET/CT shows high accuracy estimating the nature of solid LN. SUVmax ≤ 1,0 is highly predictive of benignity, and SUVmax ≥ 2,5 is highly predictive of malignancy. SUVmax values between 1,0 and 2,5 were not able to characterize efficiently LN.
Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Idoso , Idoso de 80 Anos ou mais , Adulto Jovem , Nódulo Pulmonar Solitário/diagnóstico por imagem , Tomografia por Emissão de Pósitrons combinada à Tomografia Computadorizada/métodos , Neoplasias Pulmonares/diagnóstico por imagem , Valor Preditivo dos Testes , Sensibilidade e Especificidade , Nódulo Pulmonar Solitário/patologia , Neoplasias Pulmonares/patologiaRESUMO
BACKGROUND/OBJECTIVES: Obesity is related to a dynamic extracellular matrix (ECM) remodeling, which involves the synthesis and degradation of different proteins, such as tenascin C (TNC) in the adipose tissue (AT). Given the functional relationship between leptin and inducible nitric oxide synthase (iNOS), our aim was to analyze the impact of the absence of the iNOS gene in AT inflammation and ECM remodeling in ob/ob mice. SUBJECTS/METHODS: The expression of genes involved in inflammation and ECM remodeling was evaluated in 10-week-old male double knockout (DBKO) mice simultaneously lacking the ob and iNOS genes as well as in ob/ob mice classified into three groups [control, leptin-treated (1 mg kg-1 day-1) and pair-fed]. RESULTS: Leptin deficiency increased inflammation and fibrosis in AT. As expected, leptin treatment improved the obesity phenotype. iNOS deficiency in ob/ob mice improved insulin sensitivity, AT inflammation, and ECM remodeling, as evidenced by lower AT macrophage infiltration and collagen deposition, a downregulation of proinflammatory and profibrogenic genes Tnf, Emr1, Hif1a, Col6a1, Col6a3, and Tnc, as well as lower circulating TNC levels. Interestingly, leptin upregulated TNC expression and release in 3T3-L1 adipocytes, and iNOS knockdown in 3T3-L1 fat cells produced a significant decrease in basal and leptin-induced Tnc expression. CONCLUSIONS: Ablation of iNOS in leptin-deficient mice improved AT inflammation and ECM remodeling-related genes, attenuating fibrosis, and metabolic dysfunction. The activation of iNOS by leptin is necessary for the synthesis and secretion of TNC in adipocytes, suggesting an important role of this alarmin in the development of AT inflammation and fibrosis.
Assuntos
Inflamação/metabolismo , Leptina/genética , Óxido Nítrico Sintase Tipo II/genética , Obesidade/metabolismo , Tenascina/metabolismo , Células 3T3-L1 , Tecido Adiposo/química , Tecido Adiposo/metabolismo , Animais , Fibrose/metabolismo , Inativação Gênica , Leptina/metabolismo , Camundongos , Camundongos Knockout , Camundongos Obesos , Óxido Nítrico Sintase Tipo II/metabolismoAssuntos
Melanoma/patologia , Doenças da Unha/patologia , Neoplasias Cutâneas/patologia , Adulto , Feminino , HumanosRESUMO
BACKGROUND/OBJECTIVES: Glycerol is a key metabolite for lipid accumulation in insulin-sensitive tissues as well as for pancreatic insulin secretion. We examined the role of aquaporin-7 (AQP7), the main glycerol channel in ß-cells, and AQP12, an aquaporin related to pancreatic damage, in the improvement of pancreatic function and steatosis after sleeve gastrectomy in diet-induced obese rats. SUBJECTS/METHODS: Male Wistar obese rats (n=125) were subjected to surgical (sham operation and sleeve gastrectomy) or dietary (pair-fed to the amount of food eaten by sleeve-gastrectomized animals) interventions. The tissue distribution and expression of AQPs in the rat pancreas were analyzed by real-time PCR, western blotting and immunohistochemistry. The effect of ghrelin isoforms and glucagon-like peptide 1 (GLP-1) on insulin secretion, triacylglycerol (TG) accumulation and AQP expression was determined in vitro in RIN-m5F ß-cells. RESULTS: Sleeve gastrectomy reduced pancreatic ß-cell apoptosis, steatosis and insulin secretion. Lower ghrelin and higher GLP-1 concentrations were also found after bariatric surgery. Acylated and desacyl ghrelin increased TG content, whereas GLP-1 increased insulin release in RIN-m5F ß-cells. Sleeve gastrectomy was associated with an upregulation of AQP7 together with a normalization of the increased AQP12 levels in the rat pancreas. Interestingly, ghrelin and GLP-1 repressed AQP7 and AQP12 expression in RIN-m5F ß-cells. AQP7 protein was negatively correlated with intracellular lipid accumulation in acylated ghrelin-treated cells and with insulin release in GLP-1-stimulated ß-cells. CONCLUSIONS: AQP7 upregulation in ß-cells after sleeve gastrectomy contributes, in part, to the improvement of pancreatic steatosis and insulin secretion by increasing intracellular glycerol used for insulin release triggered by GLP-1 rather than for ghrelin-induced TG biosynthesis.
Assuntos
Aquaporinas/metabolismo , Grelina/metabolismo , Peptídeo 1 Semelhante ao Glucagon/metabolismo , Células Secretoras de Insulina/fisiologia , Obesidade/cirurgia , Redução de Peso/fisiologia , Animais , Western Blotting , Modelos Animais de Doenças , Derivação Gástrica , Imuno-Histoquímica , Resistência à Insulina/fisiologia , Masculino , Obesidade/metabolismo , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase em Tempo Real , Regulação para CimaRESUMO
BACKGROUND/OBJECTIVES: Body weight, body mass index (BMI) and excess weight loss (EWL) are the most frequently used measures to analyse bariatric surgery outcomes. However, these measurements do not provide accurate information on body composition (BC) with body fat (BF), importantly determining the levels of cardiometabolic risk factors. Our aim was to analyse the evolution of BC after Roux-en-Y Gastric Bypass (RYGB) and its influence on the changes of cardiometabolic risk factors in comparison to BMI and EWL. SUBJECTS/METHODS: A group of 81 obese Caucasian patients (19 males/62 females) aged 44.9±1.3 years undergoing RYGB between January 2006 and December 2011 was prospectively followed up for a period of 3 years. BC was determined by air-displacement plethysmography. Visceral adiposity, physical activity and cardiometabolic risk factors were measured. RESULTS: BF was markedly (P<0.001) reduced after the first year, increasing progressively during the second and third years after RYGB, following a different trajectory than body weight, BMI and EWL that decreased up to the second year post surgery. Markers of glucose homeostasis decreased during the first month and continued to decrease during the first year (P<0.05), remaining stabilised or slightly increased between the second and third years following RYGB. However, markers of lipid metabolism decreased (P<0.05) markedly during the first 12 months, increasing thereafter in parallel to the changes observed in BC, with the exception of high-density lipoprotein-cholesterol, which increased progressively throughout the whole period analysed. CONCLUSIONS: The adverse switch in the changes in BC between the first and the second years after RYGB may underlie the changes observed in cardiometabolic risk factors. Tracking of adiposity during the follow-up of bariatric/metabolic surgery yields clinically relevant information to better identify patients in need of increased lifestyle advice or treatment intensification.
Assuntos
Tecido Adiposo/fisiologia , Doenças Cardiovasculares/prevenção & controle , Derivação Gástrica , Síndrome Metabólica/prevenção & controle , Obesidade Abdominal/metabolismo , Obesidade Mórbida/cirurgia , Redução de Peso/fisiologia , Tecido Adiposo/metabolismo , Adulto , Índice de Massa Corporal , Doenças Cardiovasculares/metabolismo , Doenças Cardiovasculares/fisiopatologia , Feminino , Seguimentos , Humanos , Metabolismo dos Lipídeos/fisiologia , Lipoproteínas HDL/metabolismo , Masculino , Síndrome Metabólica/metabolismo , Síndrome Metabólica/fisiopatologia , Pessoa de Meia-Idade , Obesidade Mórbida/metabolismo , Obesidade Mórbida/fisiopatologia , Pletismografia , Estudos Prospectivos , Fatores de Tempo , Resultado do Tratamento , Adulto JovemRESUMO
We introduce an image cytometer (I-CYT) for the analysis of phytoplankton in fresh and marine water environments. A linear quantification of cell numbers was observed covering several orders of magnitude using cultures of Tetraselmis and Nannochloropsis measured by autofluorescence in a laboratory environment. We assessed the functionality of the system outside the laboratory by phytoplankton quantification of samples taken from a marine water environment (Dutch Wadden Sea, The Netherlands) and a fresh water environment (Lake Ijssel, The Netherlands). The I-CYT was also employed to study the effects of two ballast water treatment systems (BWTS), based on chlorine electrolysis and UV sterilization, with the analysis including the vitality of the phytoplankton. For comparative study and benchmarking of the I-CYT, a standard flow cytometer was used. Our results prove a limit of detection (LOD) of 10 cells/ml with an accuracy between 0.7 and 0.5 log, and a correlation of 88.29% in quantification and 96.21% in vitality, with respect to the flow cytometry results.
RESUMO
BACKGROUND/OBJECTIVES: Uroguanylin and guanylin are secreted by intestinal epithelial cells as prohormones postprandially and act on the hypothalamus to induce satiety. The impact of obesity and obesity-associated type 2 diabetes (T2D) on proguanylin and prouroguanylin expression/secretion as well as the potential role of guanylin and uroguanylin in the control of lipolysis in humans was evaluated. SUBJECTS/METHODS: Circulating and gastrointestinal expression of proguanylin (GUCA2A) and prouroguanylin (GUCA2B) were measured in 134 subjects. In addition, plasma proguanylin and prouroguanylin were measured before and after weight loss achieved either by Roux-en-Y gastric bypass (RYGB) (n=24) or after a conventional diet (n=15). The effect of guanylin and uroguanylin (1-100 nmol l(-1)) on lipolysis was determined in vitro in omental adipocytes. RESULTS: Circulating concentrations of prouroguanylin, but not proguanylin, were decreased in obesity in relation to adiposity. Weight loss achieved by RYGB increased plasma proguanylin and prouroguanylin. Obese T2D individuals showed higher expression of intestinal GUCA2A as well as of the receptors of the guanylin system, GUCY2C and GUCY2D, in omental adipocytes. The incubation with guanylin and uroguanylin significantly stimulated lipolysis in differentiated omental adipocytes, as evidenced by hormone-sensitive lipase phosphorylation at Ser563, an increase in fatty acids and glycerol release together with an upregulation of several lipolysis-related genes, including AQP3, AQP7, FATP1 or CD36. CONCLUSIONS: Both guanylin and uroguanylin trigger lipolysis in human visceral adipocytes. Given the lipolytic action of the guanylin system on visceral adipocytes, the herein reported decrease of circulating prouroguanylin concentrations in obese patients may have a role in excessive fat accumulation in obesity.
Assuntos
Adipócitos/metabolismo , Hormônios Gastrointestinais/metabolismo , Gordura Intra-Abdominal/patologia , Lipólise , Peptídeos Natriuréticos/metabolismo , Redução de Peso , Adulto , Proteínas de Transporte/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Diabetes Mellitus Tipo 2/fisiopatologia , Dieta Redutora , Feminino , Derivação Gástrica , Humanos , Mucosa Intestinal/metabolismo , Obesidade/metabolismo , Obesidade/fisiopatologia , Saciação , Transdução de Sinais , Esterol Esterase/metabolismoRESUMO
We introduce a new image cytometer design for the detection of very small particulates and demonstrate its capability in water analysis. The device is a compact microscope composed of off-the-shelf components, such as a light emitting diode (LED) source, a complementary metal-oxide-semiconductor (CMOS) image sensor, and a specific combination of optical lenses that allow, through an appropriate software, Fourier transform processing of the sample volume. Waterborne microorganisms, such as Escherichia coli (E. coli), Legionella pneumophila (L. pneumophila) and phytoplankton, are detected by interrogating the volume sample either in a fluorescent or label-free mode, i.e. with or without fluorescein isothiocyanate (FITC) molecules attached to the micro-organisms, respectively. We achieve a sensitivity of 50 cells per ml, which can be further increased to 0.2 cells per ml by pre-concentrating an initial sample volume of 500 ml with an ad hoc fluidic system. We also prove the capability of the proposed image cytometer of differentiating microbiological populations by size with a resolution of 3 µm and operating in real contaminated water.
Assuntos
Escherichia coli/isolamento & purificação , Citometria por Imagem/instrumentação , Legionella pneumophila/isolamento & purificação , Microbiologia da Água , Desenho de Equipamento , Fluoresceína-5-Isotiocianato/análise , Corantes Fluorescentes/análise , Citometria por Imagem/economia , Microscopia/instrumentação , SemicondutoresRESUMO
OBJECTIVE: Fibroblast growth factor (FGF)-21, and possibly FGF19, protect against type 2 diabetes mellitus (T2DM) and obesity in rodents. We investigated the circulating levels of FGF21 and FGF19 in obese patients with varying degrees of abnormal glucose homeostasis, and we determined gene expression for FGF receptors (FGFR1-4) and the co-receptor ß-Klotho, in liver and adipose tissues. SUBJECTS AND METHODS: We analyzed 35 lean healthy (71% men) and 61 obese patients (49% men, median body mass index (BMI): 40.5 kg m(-2), interquartile range: 34.7-46.2). Among obese patients, 36 were normoglycemic, 15 showed impaired glucose tolerance and 10 had T2DM. Biopsies from liver and visceral and subcutaneous fat from a subset of obese patients and controls were analyzed. FGF19 and FGF21 levels were measured using enzyme-linked immunosorbent assay, and tissue mRNA and protein levels by reverse transcription-polymerase chain reaction and immunoblotting. RESULTS: FGF21 serum levels were significantly increased in obese patients compared with controls (P<0.001), whereas FGF19 levels were decreased (P < 0.001). FGF21 levels were positively correlated with homeostasis model assessment of insulin resistance (P = 0.0002, r = 0.37) and insulin (P = 0.001, r = 0.32), whereas FGF19 levels were negatively correlated (P = 0.007, r = -0.27; P=0.003, r = -0.28; respectively). After adjusting for BMI, the correlations of FGF21 and FGF19 levels with indicators of abnormal glucose homeostasis were not significant. In obese patients, the hepatic expression of FGF21 was increased. (P = 0.04). ß-Klotho transcript levels in visceral fat (P = 0.002) and ß-Klotho protein levels in subcutaneous (P = 0.03) and visceral fat (P = 0.04) were significantly reduced in obese patients, whereas hepatic levels for ß-Klotho (P = 0.03), FGFR1 (P = 0.04) and FGFR3 (P = 0.001) transcripts were significantly increased. CONCLUSIONS: Obesity is characterized by reciprocal alterations in FGF19 (decrease) and FGF21 (increase) levels. Although worsened in diabetic obese patients, obesity itself appears as the predominant determinant of the abnormalities in FGF21 and FGF19 levels. Opposite changes in ß-Klotho expression in fat and liver indicate potential tissue-specific alterations in the responsiveness to endocrine FGFs in obesity.
Assuntos
Tecido Adiposo/metabolismo , Fatores de Crescimento de Fibroblastos/metabolismo , Fígado/metabolismo , Proteínas de Membrana/metabolismo , Obesidade/metabolismo , Receptores de Fatores de Crescimento de Fibroblastos/metabolismo , Magreza/metabolismo , Adulto , Índice de Massa Corporal , Feminino , Glucose , Teste de Tolerância a Glucose , Humanos , Resistência à Insulina , Proteínas Klotho , Masculino , Transdução de Sinais , EspanhaRESUMO
UNLABELLED: BACKGROUND/OBJETIVES: Obese leptin-deficient ob/ob mice exhibit high adiposity and reduced muscle mass with leptin replacement promoting weight loss and inducing muscle accretion through PGC-1α-dependent mechanisms. Our aim was to analyze in vivo and in vitro the effect of leptin on FNDC5, a novel PGC-1α-dependent myokine that is synthesized and cleaved to form irisin that induces white adipose browning. METHODS/RESULTS: Twelve-week-old male wild-type and ob/ob mice were divided in three groups as follows: control, leptin-treated (1 mg kg(-1) day(-1)) and pair-fed. Leptin administration was associated with increased gastrocnemius weight and cell surface area, higher Pgc1a and Fndc5 transcript levels and a slight increase in circulating irisin. Leptin upregulated Fndc5 expression through nitric oxide (NO)-dependent mechanisms in murine C2C12 myocytes and stimulated both basal and irisin-stimulated myogenesis, as evidenced by increased myocyte cell proliferation, higher myogenin and myonectin transcript levels together with lower mRNA expression of myostatin and dystrophin and the muscle atrophy-related factors MuRF1 and MAFbx. Interestingly, leptin downregulated Fndc5 expression in a NO-independent manner in murine differentiated subcutaneous adipocytes. Furthermore, leptin prevented the irisin-induced upregulation of both brown (Ucp1 and Cidec) and beige (Tmem26) adipocyte-specific genes and the increase in uncoupling protein-1-positive cells. CONCLUSIONS: Taken together, our results provide evidence for a regulatory role of leptin on FNDC5/irisin, favoring muscle accretion but reducing fat browning.
Assuntos
Tecido Adiposo Marrom/metabolismo , Fibronectinas/metabolismo , Leptina/metabolismo , Músculo Esquelético/patologia , Óxido Nítrico/metabolismo , Obesidade/patologia , Pigmentos Biológicos/metabolismo , Gordura Subcutânea Abdominal/patologia , Células 3T3-L1 , Tecido Adiposo Marrom/patologia , Animais , Células Cultivadas , Expressão Gênica , Leptina/administração & dosagem , Leptina/farmacologia , Masculino , Camundongos , Músculo Esquelético/metabolismo , Obesidade/metabolismo , Gordura Subcutânea Abdominal/metabolismo , Fatores de Transcrição/metabolismo , Regulação para Cima , Redução de PesoRESUMO
OBJECTIVE: Recent studies linked circulating pigment epithelium-derived factor (PEDF) to obesity-associated insulin resistance, but the main source of circulating PEDF is unknown. We aimed to investigate liver and adipose tissue PEDF gene expression in association with obesity and insulin resistance. DESIGN, SUBJECTS AND METHODS: Three (two cross-sectional and one longitudinal) independent cohorts have been studied, for adipose tissue (n=80 and n=30) and liver gene expression (n=32 and n=14). Effects of high glucose and cytokines on HepG2 cell line were also investigated. PEDF gene expression and circulating PEDF were analyzed using real-time PCR and ELISA, respectively. RESULTS: In a first cohort of subjects, PEDF relative gene expression was higher in subcutaneous (SC) than in omental (OM) adipose tissue (P<0.0001) being also higher in mature adipocytes compared with stromo-vascular cells (P<0.0001). However, OM PEDF relative gene expression was decreased in morbidly obese subjects (P=0.01). Both OM PEDF and OM PEDF receptor (PEDFR) correlated positively with lipogenic and lipolytic genes, and with genes implicated in the lipid vacuole formation. Circulating PEDF levels were not associated with fat PEDF gene expression. In the second cohort, SC PEDF was decreased in subjects with type 2 diabetes and did not change significantly after weight loss. We next explored circulating PEDF in association with markers of liver-related insulin resistance injury (alanine aminotransferase, r=0.59, P=0.001). Interestingly, liver PEDF gene expression increased with obesity and insulin resistance in men, being significantly associated with fasting glucose and glycated hemoglobin in two independent cohorts. In fact, high glucose led to increased PEDF in HepG2 cells, while inflammatory stimuli present in the adipose tissue environment downregulated PEDF. CONCLUSION: Liver, but not adipose tissue, might be the source of increased circulating PEDF linked to insulin resistance.
Assuntos
Tecido Adiposo/metabolismo , Proteínas do Olho/metabolismo , Resistência à Insulina , Fígado/metabolismo , Fatores de Crescimento Neural/metabolismo , Obesidade Mórbida/metabolismo , Serpinas/metabolismo , Adipócitos , Adulto , Índice de Massa Corporal , Diferenciação Celular , Células Cultivadas , Estudos Transversais , Ensaio de Imunoadsorção Enzimática , Proteínas do Olho/genética , Feminino , Células Hep G2 , Humanos , Resistência à Insulina/genética , Estudos Longitudinais , Masculino , Fatores de Crescimento Neural/genética , Obesidade Mórbida/epidemiologia , Obesidade Mórbida/genética , Reação em Cadeia da Polimerase em Tempo Real , Serpinas/genética , Espanha/epidemiologiaRESUMO
Adipose tissue responds dynamically to alterations in nutrient excess through adipocyte hypertrophy and hyperplasia, followed by increased angiogenesis, immune cell infiltration, extracellular matrix (ECM) overproduction, and thus, increased production of proinflammatory adipokines during the progression of chronic inflammation. Adipose tissue remodelling is an ongoing process that is pathologically accelerated in the obese state in large part mediated by ECM proteins and proteases. The ECM is subject to major modifications by adipocytes and other cell types that are infiltrated in the adipose tissue, such as macrophages and vascular cells. In obesity, unusual expression of ECM components and fragments derived from tissue-remodelling processes can influence immune cell recruitment and activation, actively contributing to inflammation. ECM turnover requires a tightly regulated balance between the synthesis of the components and their proteolysis, mainly by fibrinolytic systems and matrix metalloproteases (MMPs). In this review, we discuss the key cellular steps that lead to adipose tissue remodelling and the main molecular mechanisms and mediators in this process. We highlight the importance of hypoxia and angiogenesis in the adipose remodelling process, as well as the cross-talk between adipocytes, macrophages and ECM components.
Assuntos
Tecido Adiposo/fisiopatologia , Matriz Extracelular/fisiologia , Obesidade/etiologia , Tecido Adiposo/irrigação sanguínea , Tecido Adiposo/patologia , Animais , Matriz Extracelular/patologia , Humanos , Hipóxia/patologia , Hipóxia/fisiopatologia , Inflamação/patologia , Inflamação/fisiopatologia , Metabolismo dos Lipídeos , Macrófagos/patologia , Macrófagos/fisiologia , Metaloproteinases da Matriz/metabolismo , Modelos Biológicos , Neovascularização Patológica , Obesidade/patologia , Obesidade/fisiopatologia , ProteóliseRESUMO
AIMS/HYPOTHESIS: Proinflammatory and proapoptotic cytokines such as TNF-α are upregulated in human obesity. We evaluated the association between ghrelin isoforms (acylated and desacyl ghrelin) and TNF-α in obesity and obesity-associated type 2 diabetes, as well as the potential role of ghrelin in the control of apoptosis and autophagy in human adipocytes. METHODS: Plasma concentrations of the ghrelin isoforms and TNF-α were measured in 194 participants. Ghrelin and ghrelin O-acyltransferase (GOAT) levels were analysed by western-blot, immunohistochemistry and real-time PCR in 53 biopsies of human omental adipose tissue. We also determined the effect of acylated and desacyl ghrelin (10 to 1,000 pmol/l) on TNF-α-induced apoptosis and autophagy-related molecules in omental adipocytes. RESULTS: Circulating concentrations of acylated ghrelin and TNF-α were increased, whereas desacyl ghrelin levels were decreased in obesity-associated type 2 diabetes. Ghrelin and GOAT were produced in omental and subcutaneous adipose tissue. Visceral adipose tissue from obese patients with type 2 diabetes showed higher levels of GOAT, increased adipocyte apoptosis and increased expression of the autophagy-related genes ATG5, BECN1 and ATG7. In differentiating human omental adipocytes, incubation with acylated and desacyl ghrelin reduced TNF-α-induced activation of caspase-8 and caspase-3, and cell death. In addition, acylated ghrelin reduced the basal expression of the autophagy-related genes ATG5 and ATG7, while desacyl ghrelin inhibited the TNF-α-induced increase of ATG5, BECN1 and ATG7 expression. CONCLUSIONS/INTERPRETATION: Apoptosis and autophagy are upregulated in human visceral adipose tissue of patients with type 2 diabetes. Acylated and desacyl ghrelin reduce TNF-α-induced apoptosis and autophagy in human visceral adipocytes.
Assuntos
Aciltransferases/metabolismo , Apoptose/fisiologia , Autofagia/fisiologia , Grelina/sangue , Gordura Intra-Abdominal/enzimologia , Fator de Necrose Tumoral alfa/sangue , Acilação/fisiologia , Aciltransferases/genética , Células Cultivadas , Diabetes Mellitus Tipo 2/metabolismo , Feminino , Grelina/genética , Humanos , Gordura Intra-Abdominal/citologia , Masculino , Pessoa de Meia-Idade , Obesidade/metabolismo , Omento/citologia , Omento/enzimologia , RNA Mensageiro/metabolismoRESUMO
BACKGROUND: Sleeve gastrectomy (SG) has been used as a multipurpose surgical procedure for the treatment of morbid obesity. The aim of the study was to analyze gastric morphology and histology at two different time points after SG in rats. METHODS: Thirty-five male Wistar rats were fed ad libitum during 3 months on a high-fat diet to induce obesity. Subsequently, 25 diet-induced obese rats underwent either SG (n = 12) or a sham operation (n = 13). The remaining ten obese animals encompassed the nonoperated control group (Co). Four weeks postoperatively, 15 rats (n = 5 rats/experimental group) were sacrificed, while the remaining 20 rats were sacrificed after 16 weeks (animals/group; Co = 5, sham = 8, SG = 7) to compare the gastric morphological and histopathological changes over time. Body weight and food intake were regularly recorded. RESULTS: For both time periods, the Co groups exhibited the highest body weight, while the rats undergoing the SG showed the lowest weight gain (P < 0.05). Initially, significant differences (P < 0.005) in food intake relative to body weight were observed between the Co rats and animals undergoing surgery, which disappeared thereafter. The actual total stomach size after both experimental periods in the SG group was similar to that of non- and sham-operated rats mainly due to a forestomach enlargement, which was more pronounced after 16 weeks. Traits of gastritis cystica profunda characterized by gastric foveolae elongation with hyperplasia and cystic dilatation of the glands were observed in the residual stomachs of the sleeve-gastrectomized rats. These findings were mostly observed after 16 weeks of performing the SG, although they were also detected occasionally following 4 weeks postoperatively. No intestinal metaplasia was observed. CONCLUSION: After SG gastric macro- and microscopic changes with functional implications in both the short and long term take place.
Assuntos
Gastrectomia , Obesidade Mórbida/patologia , Obesidade Mórbida/cirurgia , Estômago/patologia , Animais , Dieta Hiperlipídica , Modelos Animais de Doenças , Gastrectomia/métodos , Imuno-Histoquímica , Masculino , Obesidade Mórbida/etiologia , Ratos , Ratos Wistar , Estômago/cirurgia , Fatores de Tempo , Redução de PesoRESUMO
BACKGROUND AND AIMS: Nicotinamide phosphoribosyltransferase (NAMPT) is an adipokine with physiological effects on the control of glucose homeostasis as well as potentially involved in inflammation. The association of circulating NAMPT concentrations with obesity has not been clearly established. The aim of the present work was to evaluate the effect of obesity on circulating concentrations and gene expression levels of NAMPT in human peripheral blood cells (PBCs) as well as its involvement in inflammation, glucose and lipid metabolism. METHODS AND RESULTS: Forty-four serum samples obtained from 14 lean and 30 obese volunteers were used to analyse the circulating concentrations of NAMPT. In addition, PBC, omental adipose tissue (OM) and liver biopsy samples obtained from a subgroup of subjects were used to determine transcript levels of NAMPT by Real-time PCR. Glucose and lipid profile as well as several inflammatory factors and hepatic enzymes were analysed. NAMPT circulating concentrations (P<0.01) and gene expression levels in PBC (P<0.05) were significantly increased in obese patients as compared to lean subjects. Total-cholesterol (P=0.016), HDL-cholesterol (P=0.036) and triglycerides (P=0.050) were significant and independent determinants of circulating concentrations of NAMPT (P<0.01). Moreover, a positive correlation (P<0.01) was found with the hepatic enzymes alanine aminotransferase, aspartate aminotransferase, and γ-glutamyltransferase after BMI adjustment. CONCLUSION: Our work shows that NAMPT circulating concentrations and mRNA expression levels in PBC are increased in obese patients and that plasma NAMPT levels are related to inflammation, lipid metabolism and hepatic enzymes suggesting a potential involvement in fatty liver disease and in the obesity-associated inflammatory state.