Made available in DSpace on 2016-06-02T19:22:51Z (GMT). No. of bitstreams: 1 2555.pdf: 842616 bytes, checksum: 8ea895d65abda1d2a1c558d27226a50b (MD5) Previous issue date: 2009-07-31 / Financiadora de Estudos e Projetos / Myostatin (MSTN) is a secreted growth factor expressed in skeletal muscle and negatively regulates skeletal muscle mass. MSTN is also expressed in adipose tissue. Recent data suggest that MSTN regulates adipogenesis and energy metabolism. Aims: The objective of this study was to determine the influence of exercise in the expression of myostatin (MSTN) and its receptor in fat and muscle in obese and insulin resistant rats. Methods: Adult male Wistar rats were housed under controlled conditions (20-220 C, 10-14h light-dark cycle) and were allowed free access to standard rodent chow (control group, CG) or HF diet (58% Kcal from fat, high-fat group, HG) during 12 weeks. Glucose tolerance test (GTT) and insulin tolerance test (ITT) were performed before and at 3, 4, 8 and 12 weeks after diet regimen. After 12 weeks, CG and HG rats were randomly assigned to a swimming training group (CGE and HGE) or a sedentary group (CGS and HGS). CGE and HE swam individually in water tanks (50x30cm) at 340 C, for 45 minutes at 0900h and 1700h, 5 day week-1, for 4 weeks. After this period, rats were submitted again to GTT and ITT and then killed by decapitation. GTT area under the curve was lower in HE compared to HGS. White gastrocnemius muscle and fat pads were dissected, weighted, immediately cooled in liquid nitrogen and stored at - 700 C for subsequent analysis. The mRNA levels were quantified by real time RT-PCR. The animals were maintained according to the local University Committee guidelines for the care and use of laboratory animals. Results: HG developed insulin resistance according to GTT and ITT tests. Adipose fat pads (epididymal, retropritoneal and mesenteric) were 2 to 3 times heavier in high-fat fed groups (HGS and HGE) compared to standard chow fed groups (CGS and CGE). In HGS, the expression of MSTN was significantly higher in mesenteric fat and lower in epididymal fat and Brown tissue adipose (BAT) compared to CGS rats. The expression of ActRIIB significantly decreased in epididymal fat and BAT. The training resulted in decreased of expression of expression of MSTN in muscle and mesenteric fat and increased expression in BAT of CGE compared CGS. The expression of ActRIIB increased significantly in mesenteric fat and BAT in CGE compared to CGS. The expression of MSTN decreased in mesenteric fat and increased in epididymal fat and BAT in HGE compared to HGS. The expression of ActRIIB decreased in muscle and mesenteric fat and increased in BAT in HGE compared to HGS. Conclusions: These data suggest that the expression of MSTN in fat tissues may be involved in the metabolic response of rats submitted to high fat diet and exercise. / Miostatina (MSTN) é um fator de crescimento expresso e secretado pelo músculo esquelético que regula negativamente a massa muscular. MSTN é também expressa no tecido adiposo em menores quantidades. Dados recentes sugerem que a MSTN regula a adipogênese e o metabolismo energético. Objetivo: Verificar o efeito da dieta hiperlipídica e exercício na expressão da MSTN e de seu receptor (ActRIIB) no músculo esquelético e tecido adiposo na obesidade dietética e resistência insulínica. Métodos: Ratos Wistar adultos realizaram testes de tolerância à glicose (TTG) e tolerância à insulina (TTI) antes e após 3, 4, 8 e 12 semanas sob alimentação com dieta padrão (Grupo Controle, GC) ou dieta hiperlipídica (Grupo dieta hiperlipídica, GH). Após 12 semanas, os ratos dos GC e GH foram, aleatoriamente, separados nos grupos Controle Exercício (GCE), Dieta Hiperlipídica Exercício (GHE), Controle Sedentário (GCS) e Dieta Hiperlipídica Sedentário (GHS). GCE e GHE nadaram em tanques individuais (50 x 30cm) a 34ºC, por 45 minutos, às 9h e 17h, 5 dias semana, durante 4 semanas. Após esse período, os ratos foram submetidos à TTG e sacrificados por decapitação. Músculo Gastrocnêmio e tecido adiposo foram dissecados, pesados, imediatamente congelados e estocados a -80ºC. O RNAm da MSTN e ActRIIB foi quantificado por RT-PCR. Resultados: GH desenvolveu resistência insulínica de acordo com TTG e TTI. O treinamento aumentou a sensibilidade à insulina no GHE. No GHS, a expressão de MSTN foi significativamente maior na gordura mesentérica e menor na gordura epididimal e tecido adiposo marrom (TAM) comparado ao GCS. A expressão de ActRIIB diminuiu significativamente na gordura epididimal e TAM do GHS comparada ao GCS. O treinamento resultou em redução significativa da expressão de MSTN no músculo e gordura mesentérica e aumento no TAM do GCE comparado ao GCS. A expressão de ActRIIB diminuiu significativamente na gordura mesentérica e TAM no GCE comparado ao GCS. A expressão de MSTN diminuiu na gordura mesentérica e aumentou na gordura epididimal e TAM no GHE comparada ao GHS. A expressão de ActRIIB diminuiu no músculo e gordura mesentérica e aumentou no TAM no GHE comparado ao GHS. Conclusão: Esses dados sugerem que a expressão de MST e ActRIIB no tecido adiposo pode estar envolvida na resposta metabólica de ratos submetidos à dieta hiperlipídica e exercício.

La myostatine, un facteur de croissance de la famille des TGF-β dont la voie de signalisation agit via l'Activine récepteur de type IIB (ActRIIB), a été identifié comme un régulateur négatif important de la croissance du muscle squelettique. Toutefois, son effet sur le métabolisme énergétique musculaire et sur la fonction du muscle reste largement inexploré. Dans mes travaux de thèse, j'ai étudié la conséquence de l'inhibition de la voie de signalisation ActRIIB sur le métabolisme musculaire, et ceci dans deux modèles expérimentaux, i) les souris constitutives knock-out myostatine et ii) après l'administration pharmacologique de l'ActRIIB soluble chez les souris adultes. Nos résultats démontrent que les souris knock-out myostatine développent une forte fatigabilité, une diminution de la respiration mitochondriale et une signature moléculaire qui tend vers un métabolisme glycolytique. Comme ces résultats peuvent s'expliquer par une conversion congénitale vers des fibres musculaires glycolytiques rapides chez ces souris, j'ai étudié l'effet de l'inhibition de la voie de signalisation ActRIIB chez la souris adulte. J'ai fourni des preuves, notamment pour la souris mdx, modèle animal de la myopathie de Duchenne, que l'inhibition de l'ActRIIB, malgré une distribution de typage de fibres qui reste normale, conduit à une intolérance extrême à l'exercice. Cela a été associé à une augmentation pathologique des taux de lactate sérique ainsi que des caractéristiques prononcées de la myopathie. Plus en détail, l'analyse biochimique et moléculaire montre que l'inhibition de la voie de signalisation ActRIIB diminue l'expression de la protéine porine, réduit la capillarisation musculaire et provoque une déficience de la phosphorylation oxydative. Je montre aussi que l’ActRIIB régule les composants clés du métabolisme musculaire, comme PPARß, Pgc1α, et PDK4, optimisant ainsi les différentes composantes du métabolisme énergétique musculaire. En somme, mes résultats démontrent que l’inhibition de l’ActRIIB provoque une myopathie métabolique, en particulier dans le contexte d’un muscle dystrophique, chez lequel un stress métabolique sous-jacent existe déjà. En conclusion, je ne peux pas recommander l'utilisation de l’inhibition de la voie de signalisation de l’ActRIIB comme stratégie thérapeutique pour les maladies musculaires. / Myostatin, a growth factor of the TGF-β family that signals through the activin receptor-IIB (ActRIIB), has been identified as an important negative regulator of skeletal muscle growth. However, its effect on muscle energy metabolism and energy dependent muscle function remains largely unexplored. I here investigated the consequence of impaired ActRIIB signaling for muscle metabolism in two experimental models, i) the constitutive myostatin knockout mice and ii) following pharmacological administration of soluble ActRIIB in adult mice. Our results demonstrate that myostatin knockout mice develop a strong fatigability, a decrease in mitochondrial respiration and a molecular signature towards a glycolytic metabolism. As these findings may be explained by the congenital shift towards fast glycolytic muscle fibers in these mice, I investigated the effect of inhibition of ActRIIB signaling in adult mice. I provide evidence, notably for the mdx mouse, model for Duchenne muscular dystrophy, that ActRIIB blockade, despite an unchanged fiber type distribution, leads to extreme exercise intolerance. This was associated with pathologically increased serum lactate levels and myopathic features. In-depth biochemical and molecular analysis demonstrates that blockade of ActRIIB signaling down-regulates the ATP channel porin, reduces muscle capillarization and leads to a consecutive deficiency in oxidative phosphorylation. I also show that ActRIIB regulates key determinants of muscle metabolism, such as Pparβ, Pgc1α, and Pdk4, thereby optimizing different components of muscle energy metabolism. Taken together, my results demonstrate that ActRIIB blockade provokes a metabolic myopathy, especially in the context of dystrophic muscle, in which an underlying metabolic stress already exists. In conclusion, I cannot recommend the use of ActRIIB signaling blockade as a therapeutic strategy for muscle diseases.