Depuis des décennies, l’un des enjeux des sciences de l’espace est d’accompagner les astronautes lors de leur séjour de courte ou longue durée, principalement en vols orbitaux à bord d’une station spatiale. Dans ce contexte, nos travaux participent à la compréhension du déconditionnement musculaire associé à un séjour en impesanteur et à l’évaluation des différentes contremesures.Que ce soit dans le secteur spatial ou celui de la santé, les scientifiques ont eu l’opportunité d’étudier les effets de la réduction chronique d’activité et de l’impesanteur, grâce à différents modèles expérimentaux qui sont présentés dans le chapitre 1. La partie 1 fait également l’état des lieux des paramètres du déconditionnement musculaire, que sont principalement la perte de masse et de force musculaire. Ces deux paramètres apparaissent de manière précoce dans une situation d’hypoactivité et ne sont pas corrélés au cours du temps, ce qui se traduit par une diminution de force de plus grande amplitude que les degrés d’atrophie rapportés. Parmi les facteurs explicatifs de cette disproportion, l’accumulation accrue d’infiltrations graisseuses et la dénervation sont des éléments explicatifs et ont également fait l’objet de nos expérimentations.Nos travaux ont pour objectif principal de caractériser les altérations précoces du muscle strié squelettique à l’aide d’un modèle d’impesanteur simulée, appelé immersion sèche ou Dry Immersion (DI). Ce modèle est reconnu depuis plusieurs années par les scientifiques russes pour être plus sévère que l’alitement prolongé et a été mis en place il y a quelques années pour la première fois en Europe (Toulouse). L’étude a consisté à placer 12 bénévoles masculins sains en immersion sèche pendant 3 jours. Cette première étude menée en Europe nous a permis de caractériser les changements précoces du déconditionnement musculaire au niveau structural et fonctionnel. Les biopsies musculaires pre- et post-DI, obtenues à partir du muscle Vastus Lateralis ont permis de quantifier par immunohistologie le degré d’atrophie musculaire par type de fibre, et d’analyser les changements myotypologiques. Au niveau structural, nous avons rapporté par IRM une diminution de 10% du volume du quadriceps, ainsi qu’une atrophie de 10%, principalement attribuée aux fibres de type 1. Dès 3 jours, le pourcentage de fibres exprimant des MyHC 1 a également diminué, alors qu’on relève en parallèle une apparition de fibres hybrides (+1.3%). Au niveau fonctionnel, la perte de force musculaire est de 11%, et nous relevons une altération des propriétés viscoélastique du rectus femoris. Au niveau moléculaire, l’expression de protéines clés impliquées dans la régulation de la balance protéique rapporte par exemple une augmentation précoce de l’expression de l’atrogène MURF1 (+41%) et une diminution de l’expression de 4EBP1 (-17%). Une analyse transcriptomique par RNA-seq a également mis en valeur un changement d’expression significatif de 2872 gènes.Une seconde étude nous a permis d’évaluer les effets d’un cocktail anti-oxydant, anti-inflammatoire comme contremesure lors d’une réduction d’activité de 20 jours et lors d’un bed rest de 2 mois. Des biopsies musculaires pre- et post-DI et BR ont été obtenues à partir du muscle Vastus Lateralis et rapportent un degré d’atrophie d’environ 20% dans les deux protocoles. La contremesure a principalement montré un effet protecteur sur les fibres de type Iia. L’analyse des E3 ligases MuRF-1 et Atrogin-1, n’indique aucune modification dans les deux protocoles de longue durée.Les résultats de nos études à courts et longs termes du déconditionnement musculaire soulignent les changements rapides et précoces au niveau du système musculaire et mettent en valeur la nécessité d’étudier finement ces changements initiaux et bio-marqueurs du déconditionnement musculaire ainsi que la nécessité d’optimiser les effets des contremesures sur cette période initiale. / For decades, one of the challenges of space science has been to assist the astronauts during their short or long term stay, mainly in orbital flights aboard a space station. In this context, our work contributes to the understanding of muscle deconditioning associated with a stay in weightlessness and to the evaluation of differents countermeasures.Whether in the space or health sector, scientists have had the opportunity to study the effects of chronic hypoactivity and weightlessness through various experimental models that are presented in Chapter 1. Part 1 also reviews the parameters of muscle deconditioning, which are mainly loss of mass and muscle strength. These two parameters appear early in a situation of hypoactivity and are not correlated with time, which results in a decrease in force of greater amplitude than the degrees of atrophy reported. Among the explanatory factors of this disproportion, the increased accumulation of fatty infiltrations and the denervation processes are explanatory elements and have also been the subject of our experiments.Our main objective is to characterize the early alterations of skeletal muscle using a model of dry immersion (DI). This model has been recognized for several years by Russian scientists to be more severe than extended bed rest and was set up a few years ago for the first time in Europe (Toulouse). The study consisted of placing 12 healthy male volunteers in dry immersion for 3 days. This first study in Europe allowed us to characterize the early changes of muscle deconditioning at the structural and functional level. The pre- and post-DI muscle biopsies, obtained from the Vastus Lateralis muscle, were used to quantify by immunohistology the degree of muscle atrophy by fiber type and to analyze the changes in myotypology. At the structural level, we reported by MRI a 10% decrease in the volume of the quadriceps, as well as a 10% atrophy, mainly attributed to type 1 fibers. From 3 days, the percentage of fibers expressing MyHC 1 also decreased, while the percentage of hybrid fiber increased (+ 1.3%). At the functional level, the loss of muscle strength is 11%, and we note an alteration of the viscoelastic properties of the rectus femoris. At the molecular level, we reported the changes of key proteins involved in protein balance regulation, for example, an early increase in the expression of the atrogene MURF1 (+ 41%) and a decrease expression of 4EBP1 (-17%). Transcriptomic analysis by RNA-seq also highlighted a significant expression change of 2872 genes.A second study allowed us to evaluate the effects of an anti-oxidant, anti-inflammatory cocktail as countermeasure during a hypoactivity experiments of 20 days and during a bed rest of 2 months. Pre- and post-DI and BR muscle biopsies were obtained from the Vastus Lateralis muscle and reported a degree of atrophy of about 20% in both protocols. The countermeasure has mainly shown a protective effect on type Iia fibers. Analysis of the E3 ligases MuRF-1 and Atrogin-1 did not indicate any modification in the two long-term protocols.The results of our short and long term studies of muscle deconditioning underline the rapid and early changes in the muscle system and highlight the need to study finely these initial changes and biomarkers of deconditioning, in order to optimize the effects of countermeasures over this initial period.

Phosphatidylinositol 5-phosphate (PtdIns5P) is the least well-characterised member of the phosphoinositide family of essential regulatory phospholipids. PtdIns5P levels are altered within cells in response to a number of stimuli and evidence is accumulating to suggest that it possesses important functions in cellular signalling. However, the physiological role of this lipid remains imperfectly understood. Previous studies have shown that PtdIns5P is elevated in adipocytes in response to insulin, and microinjection of PtdIns5P into these cells promotes plasma membrane insertion of the insulin-regulated glucose transporter GLUT4 (Sbrissa et al., 2004). This finding suggests a potential role of PtdIns5P as a mediator in insulin-stimulated glucose uptake, a process essential for efficient glucose homeostasis. As approximately 75% of postprandial glucose disposal is carried out by skeletal muscle, it is important to investigate the role of PtdIns5P in the response of this tissue to insulin. Therefore, this work has used differentiated myotubes of the rat muscle cell line, L6, to explore the effects of altered PtdIns5P levels on insulin-stimulated glucose uptake. This cell model had not been previously used in the laboratory so it first required characterisation. Here insulin is shown to stimulate a transient increase of PtdIns5P in L6 myotubes, indicative of a signalling role in response to insulin. This project developed several tools to further investigate this potential role for PtdIns5P in the insulin response of myotubes. One such development was the successful overexpression of the PtdIns5P 4-kinase PIP4KIIalpha in these cells, which was able to abolish the insulin-stimulated PtdIns5P rise. This correlated with a loss of insulin-stimulated glucose uptake (upon PIP4KIIalpha expression). Interestingly, artificial elevation of PtdIns5P in L6 myotubes increases glucose uptake in the absence of stimulation. This phenomenon appears to result from the activation of PI3-kinase signalling, as it is abolished by the PI3-kinase inhibitor wortmannin, and involves activation of the PI3-kinase effector Akt. These results are consistent with the idea that insulin-stimulated PtdIns5P production contributes to the robust PI3-kinase/Akt activation necessary for insulin-stimulated glucose uptake in muscle.

Abstract Non-pathological change in body weight and adiposity is one distinct adaptive feature that seasonal species undergo, and it can offer a novel way to study the mechanisms underlying body weight regulation and energy homeostasis. Changes in the expression and activity of metabolic enzymes are essential for the physiological adaptation seasonal species exhibit. AMP-activated protein kinase (AMPK) is a key regulatory enzyme that controls the energy homeostasis both on cellular and whole-body level. In this thesis, the main focus was to clarify how seasonal adaptation affects AMPK and its downstream target in lipid metabolism, acetyl-CoA carboxylase (ACC), in different metabolic tissues of two model species with diverse wintering strategies: the raccoon dog and the Djungarian hamster. In addition, the effect of periodic fasting on the raccoon dog skeletal muscle was studied. It was observed that seasonal differences in AMPK and ACC expression were evident mainly in adipose tissues of both species. AMPK was down-regulated in white adipose tissue (WAT) of the winter-adapted raccoon dog, whereas in the Djungarian hamster WAT, the abundance of AMPK increased in response to winter acclimatization. ACC expression was maintained or increased in winter in both species. The seasonal changes in AMPK and ACC expression observed, in particular, in adipose tissues reflects the wintering strategy of the species and presumably facilitates the lipid usage and/or preservation during wintertime scarcity. Raccoon dogs were quite resistant to the prolonged wintertime fast, as no changes were observed in AMPK and ACC expression levels in the WAT, liver or hypothalamus between the fasted and fed groups. Skeletal muscle function also appears to be well preserved, as there were no changes in the expression of proteins involved in insulin signaling, and the fiber type composition and muscle energy reserves were not affected. This thesis offers novel information on protein level changes in metabolic adaptation. / Tiivistelmä Useat luonnonvaraiset eläinlajit ovat fysiologisesti sopeutuneet ravinnonsaannin vuodenaikaisiin vaihteluihin. Vuodenaikaisrytmiin kytketty rasvakudoksen määrän vaihtelu ja siihen liittyvät aineenvaihdunnalliset muutokset tarjoavat mielenkiintoisen tutkimuskohteen ruumiinpainon säätelyn ja energiatasapainon ylläpidon molekulaaristen mekanismien selvittämiseen. Oleellinen osa fysiologista sopeutumista ovat muutokset energia-aineenvaihduntaa säätelevien proteiinien ekspressio- ja aktiivisuustasoissa. Yksi keskeinen elimistön energiatasapainoa kontrolloiva entsyymi on AMP-aktivoituva proteiinikinaasi (AMPK). AMPK toimii solunsisäisenä energiasensorina ja säätelee energiametaboliaa koko kehon tasolla. Tässä väitöskirjatutkimuksessa selvitettiin talviadaptaation vaikutusta AMPK:n ja sen kohdemolekyylin, rasvahappojen biosynteesiä säätelevän asetyyli-CoA karboksylaasin (ACC), ilmenemiseen ja aktiivisuuteen eri kudoksissa. Mallieläiminä käytettiin kahta eri talvehtimisstrategian omaavaa ja eri lailla ruumiinpainoaan säätelevää lajia, kääpiöhamsteria ja supikoiraa. Lisäksi tutkittiin pitkäaikaisen talvipaaston vaikutusta supikoiran luustolihakseen. Tulokset osoittivat, että molemmilla lajeilla AMPK- ja ACC-pitoisuuksissa on vuodenaikaisia eroja erityisesti rasvakudoksessa. Supikoiralla AMPK:n määrä väheni talviadaptaation seurauksena, kun taas kääpiöhamstereilla talviakklimatisaatio johti korkeampaan AMPK-pitoisuuteen rasvakudoksissa. ACC-pitoisuus puolestaan säilyi samana tai oli korkeampi talviadaptoituneilla yksilöillä. Havaitut muutokset AMPK:n ja ACC:n ilmenemisessä kuvastavat supikoiran ja kääpiöhamsterin eroja talvehtimisessa ja havainnollistavat entsyymien oleellista osaa rasvavarastojen vuodenaikaisessa säätelyssä ja käytössä, mikä on edellytys eläinten selviämiselle yli talven niukkuuden. Lisäksi havaittiin talviadaptoituneen supikoiran olevan melko resistentti 10 viikon paastolle tutkittujen parametrien suhteen. AMPK- ja ACC-pitoisuus tai aktiivisuus ei muuttunut aineenvaihdunnallisesti oleellisissa kudoksissa (rasvakudos, maksa, hypotalamus) paasto- ja kontrolliryhmän välillä. Supikoiran lihasten toimintakyky vaikuttaisi säilyvän, sillä insuliinisignalointiin liittyvien entsyymien pitoisuus, lihasten solutyyppikoostumus tai energiavarastot eivät muuttuneet paaston myötä. Tämä tutkimus tarjoaa uutta tietoa proteiinitason muutoksista osana fysiologista sopeutumista.

La contraction musculaire est initiée par des relâchements massifs de calcium intracellulaire après stimulation par le motoneurone. Le couplage entre la stimulation neuronale et la libération de calcium dépend du complexe de relâchement du calcium (CRC), et est réalisé dans un compartiment particulier des cellules musculaires : la triade. Les triades sont formées par une apposition de trois systèmes membranaires, deux citernes terminales du réticulum sarcoplasmique qui encadrent un tubule transverse, qui est une invagination de la membrane plasmique. Toutes les protéines du CRC sont exclusivement localisées à la triade, cependant les mécanismes de formation des triades et d'adressage des protéines du CRC dans ce compartiment spécifique sont encore largement inconnus. Au niveau de la triade, des points de contacts réguliers avec les microtubules ont été observés, et la triadine, une protéine du CRC, a été proposée comme pouvant servir de lien entre les microtubules et les autres protéines du CRC.L'objectif de ce travail est d'étudier les mécanismes à l'origine de l'organisation des membranes de la triade, de la localisation des protéines du CRC, ainsi que l'implication des microtubules dans ces processus. Nous nous sommes intéressés au rôle de deux protéines associées aux microtubules, Climp63 et MAP6. Des travaux de microscopie ont également permis de caractériser le comportement de chimères fluorescentes de triadine exprimées dans des myotubes de souris différenciés en culture.Climp63 en association avec la triadine, forme un lien entre les triades et le réseau microtubulaire. Cependant, la relation fonctionnelle entre le réseau de microtubules et le relâchement de calcium reste complexe à envisager. Si l’étude d’un modèle animal montre clairement que l’absence de la protéine MAP6 affecte la force musculaire, il n’a pas été possible de montrer que ce défaut est sous-tendu par un dysfonctionnement des microtubules. L’étude de la mise en place des triades a révélé que les microtubules participent à cette organisation, en supportant la mobilité des triades en cours de positionnement dans la fibre musculaire. Enfin, nous avons pu montrer que l’adressage de la triadine à la triade se fait par une diffusion de la protéine au sein du RS, et une rétention dans la citerne terminale grâce à un mécanisme complexe, potentiellement indépendant de RYR1, et nécessitant à la fois les domaines lumenal et cytosolique de la protéine. / Muscle contraction is initiated by massive intracellular calcium releases after motoneuron stimulation. The coupling between neuronal stimulation and calcium release depends on the calcium release complex (CRC), and takes place in a very special compartment of muscle cells : the triad. Triads are formed by the apposition of three membrane systems, two terminal cisternae of the sarcoplasmic reticulum, on each side of a transverse tubule, which is an invagination of the plasma membrane. All CRC proteins are exclusively localized at the triads, however the mechanisms leading to triad formation, and CRC protein targeting at this special compartment are still largely unknown. At the triads, regular cross-talks with microtubules were observed, and triadin, one of the CRC protein, was proposed to serve as a link between microtubules and the other CRC proteins.The goal of this work is to study the mechanisms leading to the organization of triad membranes, and to triad CRC proteins targeting, as well as the involvement of microtubules in these processes. We focused our interest on the role of two microtubule-associated proteins, Climp63 and MAP6. Microscopy studies allowed us to characterize the behaviour of fluorescent triadin chimeras expressed in mouse myotubes, differentiated in culture.Climp63 when associated to triadin forms a link between triads and microtubules. However the functional relationship between microtubules and calcium releases remains complex to consider. Even if the study of an animal model shows clearly that the lack of MAP6 protein affects muscle force, we were not able to show that this defect depends on microtubule dysfunction. The study of triad set up revealed that microtubules participate in that organization, by sustaining triads mobility during positioning in the muscle cell. Finally we were able to show that triadin targeting to triads is done by diffusion of the protein in the SR membrane, and retention in the terminal cisternae thanks to a complex mechanism. This mechanism could be independant of RyR1, but needs the lumenal and cytosolic domains of the protein.

Title: Influence of alcohol on the contractile properties of skeletal muscle Objectives: To identify the impact of alcohol to contractile attribute (Tc - contraction time and Dm - maximal displacement) m. rectus femoris. Method: The diploma thesis corresponds with empirically-theoretical based study. The research has a character of a quasiexperiment. The measured participants were consisted of students (n = 8) of Military Department (VO) and students (n = 3) of civilian (TVS) of attending full-time studies at the Faculty of Physical Education and Sport (FTVS) of Charles University (UK) in Prague. There were used the methods of descriptive statistics to describe it - rate position (arithmetic mean) and a measure of variability (standard deviation). Contractile attributes (Tc, Dm) were measured on the device TGM 100 at an electricity current intensity 80 mA. To analyse the normality of data was used Kolmogorov-Smirkov's test. For further calculations was used parametric method one-way ANOVA for repeated measurements were used statistical significance. For statistical processing was used computer program IBM SPSS Statistics 22 and the individual dose of alcohol relative to body weight was administered in three rounds at 20 minute intervals. Results: The average Tc was 33.23 ± 3.45 ms for the first...

Title: The effect of intensity of voluntary isometric contraction on rheological characteristics of skeletal muscle tissue in vivo, in situ Objectives: The main aim of this study is to determine the effect of intensity of isometric voluntary contraction of skeletal muscle on its viscoelastic characteristics. The work also aims to contribute to the verification of myotonometer as an objective diagnostic instrument and compares it to known methods for evaluation of muscle tone, or its partial characteristics. Methods: We used myotonometer - utility model 29456 for evaluation of changes of stiffness and viscous behavior of skeletal muscle in vivo, in situ in 20 healthy volunteers. The values were compared during 0%, 20%, 35% and 50% of maximal strength of isometric contraction of finger flexors, controlled by hand-held dynamometer. Results: We concluded that both stiffness and viscous behavior of skeletal muscle increases with higher intensity of isometric voluntary contraction. Keywords: myotonometer, skeletal muscle, viscous behavior, stiffness, muscle tone

AN ABSTRACT OF THE DISSERTATION OF Priya Londhe, for the Doctor of Philosophy degree in Biochemistry and Molecular Biology, presented on 30th July, 2013 at Southern Illinois University Carbondale. TITLE: UNRAVELING THE MOLECULAR MECHANISMS OF THE CLASS II TRANSACTIVATOR IN SKELETAL MUSCLE MAJOR PROFESSOR: Dr. Judy Davie The inflammatory cytokine, interferon gamma, IFN-gamma orchestrates a diverse array of fundamental physiological processes and exhibits complex effects on myogenesis. IFN-gamma also induces the class II transactivator, CIITA, which is a critical mediator of IFN-gamma mediated repression and activation. The aims in my dissertation are directed towards understanding the role of IFN-gamma and CIITA in muscle. Stimulation by IFN-gamma in skeletal muscle cells induces CIITA expression as well as MHC class II gene expression. We show that the IFN-gamma induced inhibition of myogenesis is mediated by CIITA, which specifically interacts with myogenin. CIITA acts by both, repressing the expression and inhibiting the activity of myogenin at different stages of myogenesis. The IFN-gamma mediated repression is reversible, with myogenesis proceeding normally upon removal of IFN-gamma. We also show that CIITA is indispensible for the inhibition of myogenesis. To gain a mechanistic insight into the IFN-gamma induced repression of myogenesis, we have discovered that IFN-gamma and CIITA inhibit myogenesis by modifying gene regulation in a muscle cell subject to inflammation. We show that CIITA first interacts with JARID2, a non catalytic subunit of PRC2 complex, which induces a paused RNAPII, phosphorylated at serine 5 and then interacts with the catalytic subunit EZH2, in a JARID2 dependent manner. Our data show that both CIITA and IFN-gamma block myogenesis by the induction and recruitment of the PRC2 complex, which is normally silenced in a differentiating muscle cell. One of my dissertation aims sheds light on the silencing of CIITA in Rhabdomyosarcoma. Silencing of CIITA prevents the expression of MHC Class I and II genes. We have found that IFN-gamma signaling is intact in these cells, but pSTAT1 and IRF1 do not bind to the CIITA PIV promoter. The CIITA promoter is not hypermethylated in RD (ERMS) cells, but shows a modestly enhanced methylation status in SJRH30 (ARMS) cells. We have also observed that histone acetylation, which normally increases on the CIITA PIV promoter following IFN-gamma treatment, is blocked in both types of RMS cells. Further, our studies also impart a novel role for IFN-gamma and CIITA in inhibiting the IGF induced activation of muscle specific genes. Our data show that IFN-gamma does not block the signaling cascade of IGF. However, blocking exogenous IFN-gamma restores IGF activation of muscle specific genes. My dissertation also reveals an important role for the FACT complex in the early steps of gene activation through its histone chaperone activities that serve to open chromatin structure and facilitate transcription promoting muscle differentiation. We show that myogenin interacts with the FACT complex and the recruitment of FACT complex to muscle specific genes is dependent on myogenin. The final aim in my dissertation highlights the distinct binding profiles of the MRFs and E proteins during proliferation and differentiation. Our sequential ChIP assays show that MYOD, MYOG, and MYF5 co-occupy promoters. Taken together, my dissertation provides a comprehensive understanding of the molecular mechanisms during myogenesis and reveals the deleterious effects of chronic inflammation in skeletal muscle.

Le muscle strié squelettique subit de graves lésions d’ischémie-reperfusion (IR) au cours de la progression de l’artériopathie oblitérante des membres inférieurs et lors d’interventions chirurgicales qui nécessitent l’interruption transitoire du flux sanguin dans les artères des membres. Dans ce contexte, nos objectifs étaient de mettre à profit deux modèles expérimentaux d’IR des membres inférieurs par clampage aortique et garrotage unilatéral pour : ° tester l’efficacité d’une alternative médicamenteuse au postconditionnement ischémique par l’utilisation de la cyclosporine A (CsA). En se liant à la cyclophiline D, la CsA empêche l’ouverture du pore de transition mitochondrial (mPTP) à un niveau très distal de la cascade d’évènements qui conduit à la nécrose après IR. ° déterminer de quelle façon deux comorbidités fréquemment retrouvées chez des patients souffrant d’atteinte artérielle (le diabète et l’âge) influencent l’effet de la cyclosporine. Avec les protocoles de conditionnement et aux doses que nous avons utilisées, la cyclosporine a des effets différents sur les conséquences musculaires de l’ischémie-reperfusion des membres inférieurs, dépendant de la pathologie sous-jacente des animaux étudiés. Il semble intéressant d’étudier l’effet dose-réponse de la cyclosporine A pour déterminer l’intervalle thérapeutique optimal, celui-ci pouvant être différent chez l’animal sain et pathologique. D’autre part, étant donné l’importance considérable du stress oxydant chez les animaux diabétiques et sénescents, la co-administration de cyclosporine et d’un antioxydant au moment de la reperfusion pourrait rétablir une protection. / Peripheral arterial disease and many surgical procedures (requiring vascular clamping or tourniquet application) induce severe skeletal muscle ischemia-reperfusion (IR) injuries. As a result, using experimental hind limb ischemia-reperfusion models, our goals were: ° To test a pharmacologic substitute to ischemic postconditioning by using cyclosporine A, that acts on a very downstream step of IR injury cascade by binding to cyclophilin D and inhibiting mitochondrial transition pore opening. We wondered if cyclosporine could alleviate mitochondrial dysfunction and reduce ROS production in skeletal muscles submitted to IR. ° To determine how diabetes and senescence would influence cyclosporine A protective effects. In conclusion, the protective effects of pharmacologic postconditioning with cyclosporine A seem to critically depend on the model under study. A variable and narrow dose-effect relationship is likely and makes it necessary to perform a dose finding study in every pathologic model. Considering the narrow relationships between mitochondrial protection and oxidative stress, combing cyclosporine A postconditioning with antioxidant therapy may restore a more robust protective effect but this hypothesis has to be validated.

Desde seu desenvolvimento na década de 60, a fototerapia têm sido utilizada no tratamento de diversas condições patológicas, havendo um considerável corpo de evidências acerca de sua ação regenerativa, analgésica e anti-inflamatória. Tais efeitos terapêuticos podem ser explicados pela capacidade que a energia luminosa possui de ser absorvida pelos tecidos e estimular ou inibir processos intracelulares. Estudos recentes têm apresentado resultados promissores desta terapia também sobre a redução da fadiga e do dano muscular induzido pelo exercício. A fadiga muscular é um fenômeno multifacetado caracterizado por uma progressiva redução da capacidade de produção de força do músculo. O dano muscular, causado principalmente pelas ações excêntricas do exercício, é caracterizado pela desorganização da estrutura microscópica do músculo e redução da capacidade contrátil deste tecido. Assim, o objetivo do presente trabalho foi verificar o efeito da fototerapia aplicada imediatamente antes do exercício sobre: (1) a fadiga muscular de extensores de joelho submetidos a exercício isocinético concêntrico; (2) o dano muscular de extensores de joelho submetidos a exercício isocinético excêntrico. No primeiro estudo, 17 homens saudáveis e fisicamente ativos participaram de um desenho experimental cruzado no qual foram submetidos a 30 repetições concêntricas máximas de flexo-extensão do joelho, precedidas de tratamento com fototerapia ou placebo. A fototerapia foi aplicada através de um equipamento de light emitting diodes therapy (LEDT) composto por 35 diodos infravermelhos de 850 nm e 34 diodos vermelhos de 660 nm. O tratamento foi realizado em três pontos do quadríceps com aplicação de uma dose total de 125,1 J. Mensurações da função muscular dos extensores de joelho foram realizadas antes e imediatamente após o exercício através de contrações voluntárias máximas (CVM) de extensores de joelho a 60º de flexão da articulação. Como resultado, observou-se que os voluntários apresentaram um decréscimo de torque significativamente menor quando tratados com fototerapia em comparação ao tratamento placebo. No segundo estudo, 36 homens saudáveis e fisicamente ativos foram randomizados em grupo fototerapia (n=18) e grupo placebo (n=18), e submetidos a cinco séries de 15 contrações excêntricas máximas de extensores de joelho. Avaliações de dor muscular e níveis séricos das enzimas lactato desidrogenase (LDH) e creatina kinase (CK) foram mensuradas pré-exercício, 24 e 48 horas pós-exercício. Avaliações da função muscular (CVM de extensores de joelho) foram realizadas pré-exercício, imediatamente após, 24 e 48 horas após o exercício. Um equipamento de low level laser therapy (LLLT) composto por cinco diodos infravermelhos de 810 nm foi utilizado para aplicar o tratamento em seis pontos do quadríceps e transmitir uma dose total de 180 J. Como resultado, observou-se que o grupo fototerapia apresentou: (1) menores incrementos de LDH 48 horas após o exercício; (2) menores incrementos de CK 24 e 48 horas após o exercício; e (3) menor decréscimo do torque de extensores de joelho imediatamente após, 24 e 48 horas após o exercício, em comparação ao grupo placebo. Os achados destes estudos permitem concluir que o tratamento com fototerapia foi capaz de atenuar os efeitos da fadiga e do dano muscular induzidos por exercício em dinamômetro isocinético. / Since its development in the 60’s, phototherapy has been used in the treatment of several pathological conditions, with a considerable body of evidence with respect to its regenerative, analgesic and anti-inflammatory action. These therapeutic effects may be explained by the capacity that the light energy has of being absorbed by soft tissues and stimulate or inhibit intracellular processes. Recent studies have also shown promising results regarding the reduction of muscle fatigue and exercise induced muscle damage. Muscle fatigue is a multifaceted phenomenon characterized by a progressive reduction in muscle force production capacity. Muscle damage, mainly caused by eccentric exercise, is characterized by the microscopic disorganization of muscle structure and reduction of the contractile capacity of this tissue. Thus, the purpose of this study was to verify the effect of phototherapy applied immediately before exercise on: (1) knee extensors muscle fatigue after isokinetic concentric exercise; (2) knee extensor muscle damage after isokinetic eccentric exercise. In the first study 17 healthy and physically active male subjects participated of a cross-over design trial. Subjects were subjected to 30 maximal concentric repetitions of knee flexion-extension, preceded by placebo or phototherapy treatment. Phototherapy was applied with a light emitting diodes therapy (LEDT) equipment composed by 35 infrared diodes of 850 nm wavelength and 34 red diodes of 660 nm. Treatment was applied in three different points of the quadriceps muscle with a total dose of 125.1 J. Measurements of knee extensor muscle function were obtained before and immediately after exercise by maximal voluntary contractions (MVC) at a knee angle of 60º of joint flexion. Subjects showed a significant smaller decrease in torque when treated with phototherapy compared to placebo treatment. On the second study, 36 healthy and physically active male subjects were randomized into a phototherapy (n=18) and a placebo (n=18) group, and subjected to five series of 15 maximal knee extensor eccentric contractions. Measurements of pain and serum levels of lactate dehydrogenase (LDH) and creatine kinase (CK) enzymes were obtained pre-exercise, 24 and 48 hours postexercise. Evaluations of muscle function (knee extensor MVC) were obtained preexercise, immediately after, and 24 and 48 hours after exercise. A low level laser therapy (LLLT) equipment composed by five infrared diodes of 810 nm wavelength was used to apply the treatment on six different points of the quadriceps muscle with a total dose of 180 J. The phototherapy group showed: (1) smaller increments of LDH 48 hours after exercise; (2) smaller increments of CK 24 and 48 hours after exercise; and (3) smaller decrease on knee extensor torque immediately after, 24 and 48 hours after exercise compared to the placebo group. These findings allow us to conclude that the phototherapy treatment was able to attenuate the effects of fatigue and muscle damage induced by isokinetic exercise.

Made available in DSpace on 2014-06-11T19:25:36Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-02-26Bitstream added on 2014-06-13T20:33:22Z : No. of bitstreams: 1 damatto_rl_me_botfm.pdf: 863048 bytes, checksum: 20cdd00e6c2f8499693cd4baea8e854d (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A insuficiência cardíaca (IC) caracteriza-se por redução da tolerância aos exercícios com a ocorrência precoce de fadiga e dispnéia. Além de disfunção cardíaca e pulmonar, anormalidades intrínsicas da musculatura esquelética têm sido responsabilizadas pela intolerância aos esforços físicos. Em músculos periféricos e respiratórios, frequentemente são observadas atrofia e modificação nas isoformas das cadeias pesadas de miosina (MyHC) na IC. Os mecanismos e vias intracelulares de sinalização responsáveis por essas alterações ainda não estão completamente definidos. Em modelos experimentais de IC induzida por estenose aórtica ou infarto do miocárdio, verificamos que alterações na expressão dos fatores de regulação miogênica e da via miostatina/folistatina podem modular o trofismo muscular e a composição das MyHCs. Um dos modelos experimentais muito utilizados para o estudo da IC é o rato espontaneamente hipertenso (SHR). Estes animais apresentam, precocemente, hipertensão arterial e hipertrofia ventricular esquerda e, em idade avançada, desenvolvem IC. Não identificamos estudos que avaliaram o comprometimento da musculatura esquelética de SHR com IC. O objetivo deste estudo foi caracterizar as alterações da musculatura esquelética de SHR com IC por meio de avaliação da morfologia, das isoformas das cadeias pesadas de miosina e da expressão gênica e protéica dos fatores de regulação miogênica e da via miostatina/folistatina. A partir de 18 meses de idade, ratos espontaneamente hipertensos foram avaliados duas vezes por semana à procura de evidências clínicas de IC como taquipnéia, perda de peso e apatia. Após a detecção de IC, os animais foram submetidos a ecocardiograma transtorácico para a confirmação de disfunção ventricular e eutanasiados. No momento da eutanásia, foram... / Heart failure (HF) is characterized by limited exercise tolerance due to increased muscle fatigue and impaired endurance. Besides cardiac and pulmonary dysfunction, intrinsic skeletal muscle abnormalities have been shown to be involved on reduced exercise tolerance. muscle mass loss and a shift in myosin heavy chain (MyHC) isoforms have been frequently observed in peripheral and respiratory skeletal muscles during HF. The pathophysiological mechanisms and intracellular pathways responsible for muscle changes are not completely defined. We observed that myogenic regulatory factors expression and myostation/follistatin pathway modulate muscle trophism and MyHC isoforms in experimental aortic stenosis- and myocardial infarction-induced HF. The spontaneously hypertensive rat (SHR) is often used in HF studies. These rats develop systemic arterial hypertension and left ventricular hypertrophy early and HF at 18-22 month-age approximately. To the best of our knowledgement, there is not study on skeletal muscle evaluation in SHR with HF. The aim of this study was to characterize skeletal myopathy of SHR with HF by evaluating soleus muscle morphology, MyHC isoforms, and gene and protein expression of myogenic regulatory factors, myostatin, and follistatin. Eighteen month-old spontaneously hypertensive rats were evaluated twice a week to identify HF clinical features such as taquipnea, weight loss, and apathy. After detecting HF, rats were subjected to transthoracic echocardiogram. During euthanasia, we evaluated pathological evidences of HF such as pleuropericardial effusion, ascites, left atrial thrombi, right ventricular hypertrophy, and lung congestion. Agematched Wistar-Kyoto rats used as controls. Soleus morphology was analyzed in haematoxyin and eosin and picro-sirius red stained sections, and MyHC isoforms were evaluated by protein... (Complete abstract click electronic access below)