RESUMEN
BACKGROUND: Expansion of adipose tissue is dependent on adipogenesis, angiogenesis and extracellular matrix remodeling. A functional role in these processes was suggested for the gelatinase subfamily of the matrix metalloproteinases. Here, we have evaluated a potential role of gelatinase A (MMP-2) in adipogenesis. METHODS: Murine embryonic fibroblasts (MEF) were derived from wild-type or MMP-2 deficient mice. Genetic manipulation of Mmp2 (shRNA-knockdown or overexpression) was performed in 3T3-F442A preadipocytes. Cell cultures were subjected to an adipogenic medium. As an in vivo model for de novo adipogenesis, 3T3-F442A preadipocytes with or without knockdown were injected subcutaneously in Nude BALB/c mice kept on high fat diet. RESULTS: Mmp2 deficient MEF, as compared to controls, showed significantly impaired differentiation into mature adipocytes, as demonstrated by 90% reduced intracellular lipid content and reduced expression of pro-adipogenic markers. Moreover, selective Mmp2 knockdown in 3T3-F442A preadipocytes resulted in significantly reduced differentiation. In contrast, overexpression of Mmp2 resulted in markedly enhanced differentiation. In de novo formed fat pads resulting from preadipocytes with Mmp2 knockdown expression of aP2, Ppar-γ and adiponectin was significantly lower, and collagen was more preserved. The fat pad weights as well as size and density of adipocytes or blood vessels were, however, not significantly different from controls. CONCLUSION: Our data directly support a functional role of MMP-2 in adipogenesis in vitro, and suggest a potential role in in vivo adipogenesis. GENERAL SIGNIFICANCE: Selective modulation of MMP-2 levels affects adipogenesis.
Asunto(s)
Adipocitos/metabolismo , Adipogénesis/genética , Fibroblastos/metabolismo , Metaloproteinasa 2 de la Matriz/genética , Células 3T3 , Adipocitos/citología , Adipocitos/trasplante , Adiponectina/genética , Adiponectina/metabolismo , Animales , Diferenciación Celular/genética , Embrión de Mamíferos/citología , Ensayo de Inmunoadsorción Enzimática , Proteínas de Unión a Ácidos Grasos/genética , Proteínas de Unión a Ácidos Grasos/metabolismo , Fibroblastos/citología , Expresión Génica , Masculino , Metaloproteinasa 2 de la Matriz/deficiencia , Ratones , Ratones Endogámicos BALB C , Ratones Noqueados , Ratones Desnudos , Microscopía de Polarización , PPAR gamma/genética , PPAR gamma/metabolismo , Interferencia de ARN , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
BACKGROUND: CD36 is a membrane glycoprotein, contributing to the pathogenesis of metabolic disorders, like obesity, which has become a major health concern worldwide. METHODS: A potential functional role of the scavenger receptor CD36 was investigated in in vitro adipocyte differentiation and in vivo adipogenesis. RESULTS: During differentiation of 3T3-F442A preadipocytes into mature adipocytes, expression of CD36 was upregulated and CD36 gene silencing resulted in impaired differentiation, as monitored by Oil Red O staining and expression of adipogenic markers. De novo fat pad formation in NUDE mice following injection of preadipocytes was significantly reduced upon CD36 gene silencing as compared to control. This was associated with marked adipocyte hypotrophy and reduced adipose tissue adipocyte content. Macrophage infiltration in de novo fat tissues derived from preadipocytes with CD36 gene silencing was not significantly different from controls. Collagen content was significantly higher in de novo fat with CD36 gene silencing. In a nutritionally induced obesity model, total body weight as well as subcutaneous and gonadal adipose tissue mass were significantly lower in CD36 deficient mice as compared to wild-type littermates. GENERAL SIGNIFICANCE: Thus, our data support a functional role of CD36 in promoting adipogenesis in vitro as well as in vivo.
Asunto(s)
Adipocitos/citología , Adipogénesis/fisiología , Antígenos CD36/fisiología , Diferenciación Celular , Obesidad/etiología , Células 3T3 , Adipocitos/metabolismo , Animales , Peso Corporal , Antígenos CD36/química , Dieta Alta en Grasa/efectos adversos , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones Desnudos , Obesidad/prevención & controlRESUMEN
Fibrinolysis and pericellular proteolysis depend on molecular coassembly of plasminogen and its activator on cell, fibrin, or matrix surfaces. We report here the existence of a fibrinolytic cross-talk mechanism bypassing the requirement for their molecular coassembly on the same surface. First, we demonstrate that, despite impaired binding of Glu-plasminogen to the cell membrane by epsilon-aminocaproic acid (epsilon-ACA) or by a lysine-binding site-specific mAb, plasmin is unexpectedly formed by cell-associated urokinase (uPA). Second, we show that Glu-plasminogen bound to carboxy-terminal lysine residues in platelets, fibrin, or extracellular matrix components (fibronectin, laminin) is transformed into plasmin by uPA expressed on monocytes or endothelial cell-derived microparticles but not by tissue-type plasminogen activator (tPA) expressed on neurons. A 2-fold increase in plasmin formation was observed over activation on the same surface. Altogether, these data indicate that cellular uPA but not tPA expressed by distinct cells is specifically involved in the recognition of conformational changes and activation of Glu-plasminogen bound to other biologic surfaces via a lysine-dependent mechanism. This uPA-driven cross-talk mechanism generates plasmin in situ with a high efficiency, thus highlighting its potential physiologic relevance in fibrinolysis and matrix proteolysis induced by inflammatory cells or cell-derived microparticles.
Asunto(s)
Fibrinolisina/metabolismo , Fibrinólisis/fisiología , Ácido Aminocaproico/farmacología , Animales , Antifibrinolíticos/farmacología , Comunicación Celular/fisiología , Células Cultivadas , Matriz Extracelular/efectos de los fármacos , Matriz Extracelular/metabolismo , Fibrinólisis/efectos de los fármacos , Humanos , Ratones , Plasminógeno/metabolismo , Activadores Plasminogénicos/metabolismo , Procesamiento Proteico-Postraduccional , Receptor Cross-Talk/efectos de los fármacos , Receptor Cross-Talk/fisiología , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología , Activador de Plasminógeno de Tipo Uroquinasa/metabolismoRESUMEN
OBJECTIVE: The goal of this study was to examine the functional relationship between aging endothelium and thrombogenicity in a mouse model of premature aging. METHODS AND RESULTS: Coagulation tests and factors, blood cell counts, aorta endothelial function, aorta gene expression, and FeCl(3)-induced thrombosis in mesenteric blood vessels were analyzed in 10- to 30-week-old brain and muscle ARNT-like protein-1 (Bmal1)-deficient (knockout [KO]) mice and wild-type littermates. Ten-week-old KO mice manifested shortened prothrombin times (9.7 versus 11.3 seconds in wild-type) and elevated plasma fibrinogen (264 versus 172 mg/dL). At 30 weeks, factor VII (198% versus 149%), and platelet counts (2049 versus 1354 K/µL) were increased in KO mice. Gene deficiency reduced the vasoactive nitric oxide production at 10 and 30 weeks and tended to reduce and increase the protein expression of thrombomodulin and von Willebrand factor, respectively, with aging. Shortened venular and arteriolar occlusion times on FeCl(3)-induced injury in 10-week-old KO mice confirmed higher thrombogenicity, culminating in priapism, observed in 60% of 25- to 30-week-old KO males. CONCLUSION: Endothelial dysfunction and a hypercoagulable state cause early arterial and venous thrombogenicity in Bmal1 KO mice. With aging, progressive endothelial dysfunction, rising platelet counts, and high factor VII further enhance thrombogenicity, provoking priapism.
Asunto(s)
Factores de Transcripción ARNTL/deficiencia , Envejecimiento Prematuro/metabolismo , Envejecimiento Prematuro/fisiopatología , Progresión de la Enfermedad , Trombosis/metabolismo , Trombosis/fisiopatología , Factores de Transcripción ARNTL/genética , Envejecimiento/fisiología , Animales , Modelos Animales de Enfermedad , Endotelio Vascular/metabolismo , Endotelio Vascular/fisiopatología , Fibrinógeno/metabolismo , Masculino , Ratones , Ratones Noqueados , Recuento de Plaquetas , Tiempo de Protrombina , Trombomodulina/metabolismo , Factor de von Willebrand/metabolismoRESUMEN
1. Because the development of adipose tissue involves remodelling of the extracellular matrix (ECM), which requires matrix metalloproteinase (MMP) activity, we examined whether MMP inhibitors may have the potential to affect adipose tissue mass in obese mice. 2. Administration of the relatively gelatinase-specific MMP inhibitor tolylsam ((R)-3-methyl-2-[4-(3-p-tolyl-[1,2,4]oxadiazol-5-yl)-benzenesulphonylamino]-butyric acid; 100 mg/kg per day) for 7 weeks to obese wild-type mice on a high-fat diet resulted in significantly lower bodyweight (P < 0.05), lower subcutaneous (SC) and gonadal (GON) adipose tissue mass (both P < 0.05) and smaller adipocytes in both SC (P < 0.005) and GON (P < 0.0005) adipose tissues. 3. Magnetic resonance imaging confirmed a lower total body fat content in tolylsam-treated mice (P < 0.0005). In addition, tolylsam treatment of wild-type mice was associated with a marked enhancement in metabolic rate. 4. Electron microscopy analysis of tissue sections at the end of the 7 week feeding period revealed significantly higher collagen accumulation in the ECM of SC adipose tissues of tolylsam-treated mice (P < 0.001). 5. Thus, the relatively gelatinase-specific MMP inhibitor tolylsam has the potential to affect fat tissue growth in obese mice.
Asunto(s)
Tejido Adiposo/enzimología , Dieta Alta en Grasa , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Obesidad/enzimología , Oxadiazoles/farmacología , Sulfonamidas/farmacología , Tejido Adiposo/efectos de los fármacos , Tejido Adiposo/patología , Animales , Peso Corporal/efectos de los fármacos , Peso Corporal/fisiología , Dieta Alta en Grasa/métodos , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Obesos , Obesidad/tratamiento farmacológico , Oxadiazoles/uso terapéutico , Sulfonamidas/uso terapéuticoRESUMEN
A low-molecular-weight receptor tyrosine kinase inhibitor, 1-(6,7-dihydro-5H-benzo(6,7)cyclohepta(1,2-c)pyridazin-3-yl)-N3-((7-pyrrolidin-1-yl)-6,7,8,9-tetrahydro-5H-benzo(7)annulene-2-yl)-1H-1,2,4-triazole-3,5-diamine (R428) with high affinity and selectivity for the growth arrest-specific protein 6 (GAS6) receptor Axl was used to study a potential role of GAS6 signaling in adiposity. In vitro, R428 caused a concentration-dependent inhibition of preadipocyte differentiation into mature adipocytes, as evidenced by reduced lipid uptake. Inhibition of Axl-mediated signaling was confirmed by reduced levels of phospho-Akt activity. In vivo, oral administration of R428 for 5 weeks to mice kept on a high-fat diet resulted in significantly reduced weight gain and subcutaneous and gonadal fat mass. This was associated with marked adipocyte hypotrophy, enhanced macrophage infiltration, and apoptosis. Thus, affecting GAS6 signaling through receptor antagonism using a low-molecular-weight Axl antagonist impairs adipocyte differentiation and reduces adipose tissue development in a murine model of nutritionally induced obesity.
Asunto(s)
Adipocitos/efectos de los fármacos , Adipogénesis/efectos de los fármacos , Tejido Adiposo/crecimiento & desarrollo , Diferenciación Celular/efectos de los fármacos , Péptidos y Proteínas de Señalización Intercelular/fisiología , Tejido Adiposo/efectos de los fármacos , Animales , Benzocicloheptenos/farmacología , Glucemia/metabolismo , Temperatura Corporal/efectos de los fármacos , Peso Corporal/efectos de los fármacos , Tamaño de la Célula/efectos de los fármacos , Células Cultivadas , Células Madre Embrionarias/efectos de los fármacos , Macrófagos/efectos de los fármacos , Masculino , Ratones , Ratones Endogámicos C57BL , Tamaño de los Órganos/efectos de los fármacos , Proteínas Tirosina Quinasas/antagonistas & inhibidores , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Transducción de Señal , Triazoles/farmacologíaRESUMEN
Tissue-type plasminogen activator (t-PA) is approved for treatment of ischemic stroke patients, but it increases the risk of intracranial bleeding (ICB). Previously, we have shown in a mouse stroke model that stromelysin-1 (matrix metalloproteinase-3 [MMP-3]) induced in endothelial cells was critical for ICB induced by t-PA. In the present study, using bEnd.3 cells, a mouse brain-derived endothelial cell line, we showed that MMP-3 was induced by both ischemic stress and t-PA treatment. This induction by t-PA was prevented by inhibition either of low-density lipoprotein receptor-related protein (LRP) or of nuclear factor-kappaB activation. LRP was up-regulated by ischemic stress, both in bEnd.3 cells in vitro and in endothelial cells at the ischemic damage area in the mouse stroke model. Furthermore, inhibition of LRP suppressed both MMP-3 induction in endothelial cells and the increase in ICB by t-PA treatment after stroke. These findings indicate that t-PA deteriorates ICB via MMP-3 induction in endothelial cells, which is regulated through the LRP/nuclear factor-kappaB pathway.
Asunto(s)
Isquemia Encefálica/enzimología , Células Endoteliales/enzimología , Proteínas Relacionadas con Receptor de LDL/metabolismo , Metaloproteinasa 3 de la Matriz/biosíntesis , Accidente Cerebrovascular/enzimología , Activador de Tejido Plasminógeno/farmacología , Animales , Isquemia Encefálica/genética , Línea Celular Transformada , Modelos Animales de Enfermedad , Inducción Enzimática/efectos de los fármacos , Humanos , Proteínas Relacionadas con Receptor de LDL/genética , Masculino , Metaloproteinasa 3 de la Matriz/genética , Ratones , FN-kappa B/metabolismo , Accidente Cerebrovascular/genéticaRESUMEN
Administration of Tolylsam, a MMP inhibitor with relative specificity for gelatinases, at a dose of 100 mg/kg/day to leptin-deficient (ob/ob) mice kept on high fat diet for 15 weeks, was associated with significantly reduced weight gain as compared to controls (p < 0.0005), resulting in lower body weight (p < 0.0005) at the end of the experiments. Food intake, physical activity and body temperature were not affected. Subcutaneous (SC) (2.9 ± 0.1g vs. 3.4 ± 0.2g in controls; p < 0.05) and gonadal (GON) (3.4 ± 0.1g vs. 3.7 ± 0.1g in controls; p = NS) fat mass were reduced by Tolylsam treatment. Reduced MMP-2 (gelatinase A) activity in adipose tissue extracts was confirmed by zymography. Mild adipocyte hypotrophy was observed in treated SC and GON adipose tissues. Blood vessel density was significantly reduced in Tolylsam treated SC (p < 0.05) and GON (p < 0.005) adipose tissues. Sirius red staining revealed comparable collagen content in both SC and GON fat of treated mice, whereas collagen disorganization (ratio thick/thin fibers) was also similar. Thus, gelatinase inhibition in mice with leptin deficiency resulted in lower body and fat pad weights, associated with mild adipocyte hypotrophy. This indicates that MMP inhibition may impair adipose tissue development independently of leptin.
Asunto(s)
Tejido Adiposo/crecimiento & desarrollo , Leptina/fisiología , Inhibidores de la Metaloproteinasa de la Matriz , Adipocitos/citología , Tejido Adiposo/enzimología , Animales , Gelatinasas/antagonistas & inhibidores , Gelatinasas/metabolismo , Leptina/deficiencia , Masculino , Metaloproteinasa 2 de la Matriz/genética , Metaloproteinasa 2 de la Matriz/fisiología , Metaloproteinasa 9 de la Matriz/genética , Metaloproteinasa 9 de la Matriz/fisiología , Ratones , Ratones Endogámicos C57BL , Tamaño de los Órganos/efectos de los fármacos , Oxadiazoles/administración & dosificación , ARN Mensajero/análisis , Sulfonamidas/administración & dosificaciónRESUMEN
We investigated the in vitro fibrinolytic properties of microplasmin, the isolated proteinase domain of plasmin, and its thrombolytic efficacy in a coronary artery thrombosis model in dogs. The amidolytic and fibrinolytic activity of recombinant microplasmin was compared with natural human plasmin. The thrombolytic efficacy of microplasmin was studied in a canine model of copper coil induced coronary artery thrombosis. Animals were randomly assigned to one of six treatment regimens, each with five animals per cohort. Four treatment groups received an intravenous bolus of microplasmin followed by an intravenous infusion of microplasmin for 1 h (1 mg/kg + 1.5 mg/kg/h with or without abciximab or 2 mg/kg + 3 mg/kg/h). In two treatment groups, microplasmin was administered intracoronary. Bolus administration was followed by a 1-h infusion if coronary flow was incompletely restored after the initial bolus administration (1 mg/kg + 1.5 mg/kg/h or 2 mg/kg + 3 mg/kg/h, respectively). The thrombolytic efficacy was documented by repeated angiographies and the coronary perfusion was assessed with the Thrombolysis in Myocardial Infarction (TIMI) grading. No significant differences between plasmin and microplasmin were observed with respect to the catalytic efficiencies towards the synthetic chromogenic substrates S-2403 or S-2444. The concentration required for 50% lysis of purified fibrin clots in 3 h, was approximately 100 nM for microplasmin compared to 20 nM for natural plasmin. Intravenous bolus administration of microplasmin restored TIMI 3 coronary flow in 0/5, 0/5, 1/5 and 2/5, respectively, whereas intracoronary bolus administration restored TIMI 3 coronary flow in 1/5 and 4/5 (1 mg/kg and 2 mg/kg, respectively) (ANOVA P < 0.05). TIMI 3 coronary flow was obtained in 0/5, 2/5, 2/5 and 3/5, respectively, during subsequent intravenous administration and in 5/5 and 4/5 in case of intracoronary administration (ANOVA P < 0.05). When compared to natural plasmin, the catalytic efficiency of microplasmin towards chromogenic substrates was similar, but the fibrinolytic potency of microplasmin towards fibrin clots was lower. Intracoronay administration of microplasmin effectively lysed coronary thrombosis.
Asunto(s)
Trombosis Coronaria/tratamiento farmacológico , Fibrinolisina/administración & dosificación , Fragmentos de Péptidos/administración & dosificación , Terapia Trombolítica/métodos , Angiografía , Animales , Modelos Animales de Enfermedad , Perros , Relación Dosis-Respuesta a Droga , Fibrinolisina/uso terapéutico , Fibrinólisis/efectos de los fármacos , Humanos , Infusiones Intravenosas , Inyecciones Intravenosas , Oligopéptidos , Fragmentos de Péptidos/uso terapéutico , Resultado del TratamientoRESUMEN
Ageing is associated with an increase in visceral obesity in men and women. Although wild-type mice with a C57Bl/6 genetic background are extensively used in studies on obesity and metabolism, little information is available on age-associated changes in their adipose tissues. We have evaluated development and composition of subcutaneous (SC) and gonadal (GON) adipose tissue in male C57Bl/6 mice at the ages of 10 weeks, 12 months or 24 months, while kept on normal chow. Total body weight as well as SC and GON fat mass significantly increased between 10 weeks and 12 months, but markedly decreased again up to 24 months of age. Adipocyte size in both fat depots and blood vessel size in GON fat followed this trend. Plasma leptin levels correlated positively with body weight and SC or GON fat mass. Both 12 and 24 months old mice displayed better insulin sensitivity as compared to 10 weeks old counterparts, reflected by significantly decreased plasma levels of insulin and/or glucose. Thus, ageing of C57Bl/6 male mice is associated with a biphasic pattern (increase up to 12 months followed by a decrease up to 24 months) of body weight, SC and GON fat mass, adipocyte and blood vessel size.
Asunto(s)
Tejido Adiposo Blanco , Adiposidad , Envejecimiento/fisiología , Tejido Adiposo Blanco/irrigación sanguínea , Tejido Adiposo Blanco/citología , Envejecimiento/sangre , Animales , Glucemia/análisis , Vasos Sanguíneos , Peso Corporal , Recuento de Células , Tamaño de la Célula , Insulina/sangre , Resistencia a la Insulina , Grasa Intraabdominal/irrigación sanguínea , Grasa Intraabdominal/citología , Leptina/sangre , Masculino , Ratones , Ratones Endogámicos C57BL , Modelos Animales , Obesidad Abdominal/fisiopatología , Grasa Subcutánea Abdominal/irrigación sanguínea , Grasa Subcutánea Abdominal/citologíaRESUMEN
We characterized the interactions between plasminogen and neurons and investigated the associated effects on extracellular matrix proteolysis, cell morphology, adhesion, signaling and survival. Upon binding of plasminogen to neurons, the plasmin formed by constitutively expressed tissue plasminogen activator (tPA) degrades extracellular matrix proteins, leading to retraction of the neuron monolayer that detaches from the matrix. This sequence of events required both interaction of plasminogen with carboxy-terminal lysine residues and the proteolytic activity of plasmin. Surprisingly, 24h after plasminogen addition, plasmin-detached neurons survived and remained associated in clusters maintaining focal adhesion kinase phosphorylation contrasting with other adherent cell types fully dissociated by plasmin. However, long-term incubation (72 h) with plasminogen was associated with an increased rate of apoptosis, suggesting that prolonged exposure to plasmin may cause neurotoxicity. Regulation of neuronal organization and survival by plasminogen may be of pathophysiological relevance, as plasminogen is expressed in the brain and/or extravasate during vascular accidents or inflammatory processes.
Asunto(s)
Supervivencia Celular/fisiología , Neuronas/fisiología , Plasminógeno/metabolismo , Ácido Aminocaproico/metabolismo , Animales , Antifibrinolíticos/metabolismo , Células Cultivadas , Corteza Cerebral/citología , Activación Enzimática , Matriz Extracelular/metabolismo , Proteínas de la Matriz Extracelular/metabolismo , Fibrinolisina/metabolismo , Quinasa 2 de Adhesión Focal/metabolismo , Ratones , Ratones Endogámicos C57BL , Neuronas/citología , Transducción de Señal/fisiología , Activador de Tejido Plasminógeno/metabolismoRESUMEN
To study the functional role of plasminogen activator inhibitor-1 (PAI-1) in obesity, the effect of its overexpression on de novo adipogenesis was evaluated in murine models in vivo. Therefore, 3T3-F442A preadipocytes expressing murine PAI-1 (mPAI-1) or control cells were injected in the back of male NUDE mice, which were fed a high-fat diet (HFD) for four weeks. De novo fat pads that formed from the PAI-1 expressing cells were larger (21 +/- 2.4 mg vs. 14 +/- 1.4 mg; p = 0.017) and showed a higher adipocyte density (373 +/- 28 mm(-2) vs. 301 +/- 12 mm(-2); p = 0.03) as compared to those formed from control cells. In a second model, male NUDE mice were injected in the tail vein with an adenoviral construct expressing mPAI-1 or with the empty vector, and three days later with 3T3-F442A cells. After four weeks of HFD, total body weight and de novo fat pad weight were comparable for both groups. Mild adipocyte hypotrophy was observed in the de novo fat pads of the PAI-1 overexpressing mice (1180 +/- 33 microm(2) vs. 1285 +/- 32 microm(2); p = 0.024), whereas the blood vessel size was significantly smaller than in controls (30 +/- 1.8 microm(2) vs. 63 +/- 3.6 microm(2); p < 0.0001). Thus, the effect of local or systemic PAI-1 (over)expression on adipocyte or blood vessel size and density of de novo formed fat pads appears to be different, and concentration-dependent. Whereas local expression resulted in larger fat pads, systemic overexpression had no effect on de novo adipogenesis, although angiogenesis appeared to be impaired.
Asunto(s)
Adipocitos/metabolismo , Adipogénesis , Tejido Adiposo/metabolismo , Serpinas/metabolismo , Células 3T3 , Adipocitos/trasplante , Tejido Adiposo/irrigación sanguínea , Tejido Adiposo/citología , Tejido Adiposo/crecimiento & desarrollo , Animales , Peso Corporal , Tamaño de la Célula , Grasas de la Dieta/administración & dosificación , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Modelos Animales , Neovascularización Fisiológica , Serpina E2 , Serpinas/genética , Transfección , Regulación hacia ArribaRESUMEN
Development of obesity is associated with substantial modulation of adipose tissue structure, involving adipogenesis, angiogenesis, and extracellular matrix remodelling. These processes require proteolytic activity, provided mainly by the fibrinolytic (plasminogen/plasmin), matrix metalloproteinase, and ADAM/ADAMTS systems. In early-stage development of adipose tissue, adipogenesis is tightly associated with angiogenesis. Thus, adipose tissue explants trigger blood vessel formation, and in turn adipose tissue endothelial cells promote pre-adipocyte differentiation. Modulation of angiogenesis and of proteolytic systems may have the potential to impair adipose tissue development.
Asunto(s)
Tejido Adiposo/irrigación sanguínea , Neovascularización Fisiológica , Obesidad/fisiopatología , Adipocitos/metabolismo , Adipoquinas/metabolismo , Tejido Adiposo/efectos de los fármacos , Tejido Adiposo/metabolismo , Inhibidores de la Angiogénesis/farmacología , Proteínas Angiogénicas/metabolismo , Animales , Fármacos Antiobesidad/farmacología , Células Endoteliales/metabolismo , Humanos , Neovascularización Fisiológica/efectos de los fármacos , Obesidad/tratamiento farmacológico , Obesidad/metabolismo , Péptido Hidrolasas/metabolismo , Inhibidores de Proteasas/farmacología , Transducción de SeñalRESUMEN
BACKGROUND: Mice deficient in the circadian clock gene BMAL1 (Brain and Muscle ARNT-like protein-1) exhibit a hypercoagulable state and an enhanced arterial and venous thrombogenicity, which aggravates with age. We investigated the effect of BMAL1 deficiency in mice at a different age on the diurnal rhythm of factors involved in coagulation and fibrinolysis. MATERIALS AND METHODS: Hepatic, cardiac and brain tissues were isolated from 10- and 25-weeks-old Bmal1-deficient (BMAL1-/-) and wild-type (BMAL1+/+) mice at ZT2 and at ZT14 to analyze the mRNA expression level of genes involved in coagulation and fibrinolysis. RESULTS: Body weight and brain weight were significantly lower in all BMAL1-/- versus BMAL1+/+ mice. Bmal1 deficiency disturbed the diurnal rhythm of plasminogen activator inhibitor-1 (PAI-1) in liver and plasma, but not in cardiac or brain tissues. BMAL1+/+ livers showed diurnal fluctuations in factor (F)VII, FVII, protein S and anti-thrombin gene expression, which were not observed in BMAL1-/- tissues. Interestingly, tissue plasminogen activator (t-PA) expression was significantly upregulated in all BMAL1-/- versus BMAL1+/+ brains at both time points. Plasma t-PA-PAI-1 complex levels were however similar for all groups. CONCLUSION: Bmal1 deficiency affected the biphasic rhythm of coagulation and fibrinolysis factors predominantly in the liver. In the brain, Bmal1-dependent control of t-PA gene expression was documented for the first time.
Asunto(s)
Factores de Transcripción ARNTL/fisiología , Ritmo Circadiano/fisiología , Hemostasis/fisiología , Factores de Transcripción ARNTL/deficiencia , Animales , Recuento de Células Sanguíneas , Barrera Hematoencefálica , Composición Corporal , Encéfalo/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Miocardio/metabolismo , Inhibidor 1 de Activador Plasminogénico/sangre , Activador de Tejido Plasminógeno/sangreRESUMEN
Butein is a plant flavonoid chalcone, with presumed anti-adipogenic properties. It was reported to impair preadipocyte differentiation, limit adipose tissue (AT) development and enhance white AT browning in rodents. In this study, we investigated the hypothesis that these effects of butein may occur via reduction of ADAMTS5 (A Disintegrin And Metalloproteinase with ThromboSpondin motifs 5) expression. Murine 3T3-L1 or 3T3-F442A preadipocytes were differentiated into mature adipocytes in the presence of butein or vehicle. At regular time intervals RNA was collected for gene expression studies. Male hemizygous mice for Tg(Ucp1-luc2,-tdTomato)1Kajim (ThermoMouse) were exposed to butein or vehicle, after which ATs were analyzed for Adamts5 and uncoupling protein-1 (Ucp-1) mRNA level changes. During preadipocyte differentiation, butein (25 - 50 mM) did not affect Adamts5 or Ucp-1 expression. Oil Red O analysis and monitoring of differentiation markers failed to demonstrate effects of butein on the differentiation extent. Furthermore, butein administration to the ThermoMouse (10 or 20 mg/kg, 4 days) or to the C57BL6/Rj mice (20 mg/kg, 4 weeks) did not enhance Adamts5 or Ucp-1 expression. Thus, we could not demonstrate marked effects of butein on the preadipocyte differentiation extent or AT development and browning, nor on Adamts5 or Ucp-1 gene expression during these processes.
Asunto(s)
Adipocitos/efectos de los fármacos , Adipogénesis , Antiinflamatorios/farmacología , Chalconas/farmacología , Células 3T3 , Proteína ADAMTS5/genética , Proteína ADAMTS5/metabolismo , Adipocitos/citología , Adipocitos/metabolismo , Animales , Células Cultivadas , Masculino , Ratones , Ratones Endogámicos C57BL , Proteína Desacopladora 1/genética , Proteína Desacopladora 1/metabolismoRESUMEN
Obesity has become a global health-threat for every age group. It is well known that young mice (10-12 weeks of age) fed a western-type diet (WD) become obese and develop higher cholesterol levels and liver steatosis whereas insulin sensitivity is reduced. Less is known, however, about the effect of a WD on advanced-age mice. Therefore, 10 week-old (young) and 22 month-old (advanced-age), male C57BL/6JRj mice were kept on either a WD or a control diet (SFD) for 15 weeks. In contrast to young mice, advanced-age mice on WD did not show a higher body weight or adipose tissue (AT)-masses, suggesting a protection against diet-induced obesity. Furthermore, plasma adiponectin and leptin levels were not affected upon WD-feeding. A WD, however, did induce more hepatic lipid accumulation as well as increased hepatic expression of the macrophage marker F4/80, in advanced-age mice. There were no significant differences in mRNA levels of uncoupling protein-1 or F4/80 in brown AT (BAT) or of several intestinal integrity markers in colon suggesting that the protection against obesity is not due to excessive BAT or to impaired intestinal absorption of fat. Thus, advanced-age mice, in contrast to their younger counterparts, appeared to be protected against diet-induced obesity.
Asunto(s)
Factores de Edad , Dieta Occidental/efectos adversos , Obesidad/metabolismo , Tejido Adiposo/metabolismo , Tejido Adiposo Pardo/metabolismo , Animales , Hígado Graso , Glucosa/metabolismo , Resistencia a la Insulina/fisiología , Leptina/metabolismo , Hígado/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Proteína Desacopladora 1/metabolismoRESUMEN
Pregnancy-induced metabolic changes are regulated by signals from an expanded adipose organ. Placental growth factor (PlGF), acting through vascular endothelial growth factor receptor-1, may be among those signals. There is a steep rise in circulating PlGF during normal pregnancy, which is repressed in gravidas who develop preeclampsia. PlGF-deficiency in mice impairs adipose vascularization and development. Here we studied young-adult PlGF-deficient (PlGF(-/-)) and wild-type mice on a high-fat diet in the nongravid state and at embryonic day (E) 13.5 or E18.5 of gestation. Litter size and weight were normal, but E18.5 placentas were smaller in PlGF(-/-) pregnancies. PlGF(-/-) mice showed altered intraadipose dynamics, with the following: 1) less blood vessels and fewer brown, uncoupling protein (UCP)-1-positive, adipocytes in white sc and perigonadal fat compartments and 2) white adipocyte hypertrophy. The mRNA expression of beta(3)-adrenergic receptors, peroxisome proliferator-activated receptor-gamma coactivator-1alpha, and UCP-1 was decreased accordingly. Moreover, PlGF(-/-) mice showed hyperinsulinemia. Pregnancy-associated changes were largely comparable in PlGF(-/-) and wild-type dams. They included expanded sc fat compartments and adipocyte hypertrophy, whereas adipose expression of key angiogenesis/adipogenesis (vascular endothelial growth factor receptor-1, peroxisome proliferator-activated receptor-gamma(2)) and thermogenesis (beta(3)-adrenergic receptors, peroxisome proliferator-activated receptor-gamma coactivator-1alpha, and UCP-1) genes was down-regulated; circulating insulin levels gradually increased during pregnancy. In conclusion, reduced adipose vascularization in PlGF(-/-) mice impairs adaptive thermogenesis in favor of energy storage, thereby promoting insulin resistance and hyperinsulinemia. Pregnancy adds to these changes by PlGF-independent mechanisms. Disturbed intraadipose dynamics is a novel mechanism to explain metabolic changes in late pregnancy in general and preeclamptic pregnancy in particular.
Asunto(s)
Tejido Adiposo/anatomía & histología , Hiperinsulinismo/genética , Proteínas Gestacionales/genética , Preñez , Tejido Adiposo/irrigación sanguínea , Tejido Adiposo/metabolismo , Animales , Metabolismo Energético/genética , Femenino , Regulación de la Expresión Génica , Edad Gestacional , Resistencia a la Insulina/genética , Masculino , Ratones , Ratones Noqueados , Tamaño de los Órganos/genética , Fenotipo , Placenta/anatomía & histología , Factor de Crecimiento Placentario , Embarazo , Complicaciones del Embarazo/genética , Proteínas Gestacionales/metabolismo , Termogénesis/genéticaRESUMEN
Obesity is a common disorder, and related diseases such as diabetes, atherosclerosis, hypertension, cardiovascular disease and cancer are a major cause of mortality and morbidity in Western-type societies. Development of obesity is associated with extensive modifications in adipose tissue involving adipogenesis, angiogenesis and extracellular matrix proteolysis. The fibrinolytic (plasminogen/plasmin) and matrix metalloproteinase (MMP) systems cooperate in these processes. A nutritionally induced obesity model in transgenic mice has been used extensively to study the role of the fibrinolytic and MMP systems in the development of obesity. These studies support a role of both systems in adipogenesis and obesity, and suggest that modulation of proteolytic activity may affect development of adipose tissue.
Asunto(s)
Adipogénesis , Tejido Adiposo/crecimiento & desarrollo , Fibrinólisis , Metaloproteinasas de la Matriz/metabolismo , Obesidad/enzimología , Proteínas ADAM/metabolismo , Adipogénesis/efectos de los fármacos , Adipogénesis/genética , Tejido Adiposo/efectos de los fármacos , Tejido Adiposo/enzimología , Animales , Fármacos Antiobesidad/farmacología , Fármacos Antiobesidad/uso terapéutico , Modelos Animales de Enfermedad , Fibrinolisina/metabolismo , Fibrinólisis/efectos de los fármacos , Fibrinólisis/genética , Inhibidores de la Metaloproteinasa de la Matriz , Metaloproteinasas de la Matriz/genética , Ratones , Obesidad/sangre , Obesidad/tratamiento farmacológico , Obesidad/genética , Plasminógeno/metabolismo , Inhibidores de Proteasas/farmacología , Inhibidores de Proteasas/uso terapéuticoRESUMEN
The effect of rofecoxib (Vioxx), a cyclooxygenase (COX)-2 inhibitor, on adipose tissue development was studied in a murine model of diet-induced obesity. Oral administration of Vioxx for six weeks (34 mg/kg/day) to C57Bl/6 mice kept on high-fat diet (n = 19) resulted in a significant reduction in total body weight (p < 0.01) and of subcutaneous (p < 0.05) and gonadal (p < 0.01) adipose tissue mass, as compared to placebo-treated animals (n = 21). There was no significant difference in food intake between both groups (2.8 +/- 0.09 vs. 3.0 +/- 0.10 g/day; p = 0.20). Administration of Vioxx resulted in reduced total cholesterol and high-density lipoprotein (HDL) cholesterol levels (p < 0.0001) and in enhanced levels of liver enzymes, as compared to place-bo. In the gonadal but not in the subcutaneous adipose tissue, adipocytes were smaller after Vioxx treatment (p < 0.05). The macrophage content was significantly lower in gonadal adipose tissues of Vioxx-treated mice (p < 0.05), but not in the subcutaneous adipose tissues. This was, however, not associated with differences in adipose tissue levels of the pro-inflammatory tumor necrosis factor (TNF)-alpha. Blood vessel size or density in either fat pad were not affected by Vioxx treatment. Thus, in a nutritionally induced murine obesity model, oral administration of Vioxx, as compared to placebo, resulted in reduced adipose tissue development, associated with lower feeding efficiency and smaller adipocytes.
Asunto(s)
Tejido Adiposo/patología , Inhibidores de la Ciclooxigenasa 2/farmacología , Lactonas/farmacología , Obesidad/tratamiento farmacológico , Obesidad/inmunología , Sulfonas/farmacología , Tejido Adiposo/inmunología , Animales , Glucemia/metabolismo , Grasas de la Dieta/farmacología , Modelos Animales de Enfermedad , Conducta Alimentaria , Inflamación/tratamiento farmacológico , Inflamación/inmunología , Inflamación/patología , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Masculino , Ratones , Ratones Endogámicos C57BL , Obesidad/patologíaRESUMEN
An FeCl(3) induced femoral arterial thrombosis model was applied to lean (47+/-1.4 g) and obese (64+/-1.7 g) mice (Swiss genetic background) in order to study the relation between obesity and thrombotic risk. As compared to lean mice, obese mice showed a significantly shorter occlusion time (9.9+/-1.0 min versus 13+/-0.5 min; p=0.04) and lower total blood flow (37+/-7.3% versus 69+/-6.7%; p=0.008). A significant negative correlation was observed between body weight and both occlusion time (r=-0.57; p=0.014) and blood flow (r=-0.57; p=0.028). Analysis of the coagulation profile revealed significantly higher levels of plasminogen activator inhibitor-1 (PAI-1), thrombin-antithrombin complex, Factor V activity and combined Factors II/VII/X activity, and moderately elevated Factor VIII activity in obese mice. The degree of arterial damage and the thrombus extension were, however, not significantly different. A significant positive correlation was observed between body weight and either PAI-1 (r=0.63; p=0.003), Factors II/VII/X levels (r=0.80; p<0.0001) or Factor V levels (r=0.65; p=0.003). Thus, this injury induced femoral artery thrombosis model in mice establishes experimentally a correlation between obesity and prothrombotic tendency.