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A Hemoxigenase (HO) é uma enzima que atua na degradação/metabolização do grupo heme gerando trêsprodutos: o monóxido de carbono (CO), a biliverdina (BVD) e o ferro ferroso (Fe2+). Uma das isoformas daHO, a Hemoxigenase-1 (HO-1) vem sendo amplamente estudada devido às suas potentes propriedadesprotetoras, desempenhando ações anti-inflamatórias, antioxidantes, antiapoptóticas e antiproliferativas,e exercendo funções citoprotetoras em diversos tipos celulares, bem como, em diferentes tipos de lesõesem modelo animal e em humanos. A HO-1 é uma enzima induzida em resposta ao estresse celular, sendoestimulada por diversos fatores como: luz ultravioleta, hipóxia, metais pesados, óxido nítrico, citocinasinflamatórias, etanol, prostaglandinas e choque térmico. Além disso, a HO-1 também é induzida emresposta ao exercício físico realizado em diferentes durações e intensidades, conferindo proteção celulare manutenção da homeostase celular pós-exercício. Na presente revisão de literatura, em uma primeiraparte serão abordadas a localização e a função da hemoxigenase no organismo e, na segunda parte, osefeitos do exercício físico sobre a HO-1 e sua citoproteção serão abordados levando-se em consideraçãoexercícios aeróbios vs. anaeróbios; exercícios aeróbios agudos (uma única sessão de exercício) realizadosem diferentes durações e intensidades; exercício aeróbio crônico treinamento físico) e exercício físico nasdoenças crônicas.

Exercise physiology has evolved as a main area of investigation, in which the central goal is to betterunderstand how the physiological systems respond to an acute bout of exercise and how these systemsadapt to different types of exercise training. For many years and until now, exercise physiology field have been grounded in the fundamentals of biology and human physiology. However, during the last century,scientific knowledge has changed our understanding of biological sciences, allowing the integration of different areas, and increasing the focus on many sub-areas like cellular and molecular investigation. Thedevelopment of new experimental techniques in the last years provided detailed information about cellstructure and function and, as a result, we could better understand not only the human body physiology,but also many diseases and their pathophysiology. Therefore, this present review intends to discuss moreabout cellular and molecular exercise physiology area, focusing on historical and methodological approaches,and highlighting the future perspectives for scientific knowledge and their practical application in healthand exercise.

Simple Summary Chronic disease-related muscle atrophy is a serious public health problem since it reduces mobility and contributes to increases in hospitalization costs. Unfortunately, there is no approved treatment for muscle wasting at present. Thus, an understanding of the mechanisms underlying the control of muscle mass and function under chronic diseases can pave the way for the discovery of innovative therapeutic strategies to counteract muscle wasting. Since numerous types of cancer induce cachexia, which has no cure nor an effective treatment, the main proposal here was to study the effects of AET in cancer cachexia, and to investigate, through in vivo manipulation of the Akt/mTORC1 pathway, whether the cachectic muscle still presents conditions to respond adaptively to hypertrophic stimuli. Our results could provide a basis for innovative research lines to better understand muscle plasticity and to investigate potential therapeutic approaches necessary to prevent muscle loss. Abstract Cancer cachexia is a multifactorial and devastating syndrome characterized by severe skeletal muscle mass loss and dysfunction. As cachexia still has neither a cure nor an effective treatment, better understanding of skeletal muscle plasticity in the context of cancer is of great importance. Although aerobic exercise training (AET) has been shown as an important complementary therapy for chronic diseases and associated comorbidities, the impact of AET on skeletal muscle mass maintenance during cancer progression has not been well documented yet. Here, we show that previous AET induced a protective mechanism against tumor-induced muscle wasting by modulating the Akt/mTORC1 signaling and eukaryotic initiation factors, specifically eIF2-α. Thereafter, it was determined whether the in vivo Akt activation would induce a hypertrophic profile in cachectic muscles. As observed for the first time, Akt-induced hypertrophy was able and sufficient to either prevent or revert cancer cachexia by modulating both Akt/mTORC1 pathway and the eIF-2α activation, and induced a better muscle functionality. These findings provide evidence that skeletal muscle tissue still preserves hypertrophic potential to be stimulated by either AET or gene therapy to counteract cancer cachexia.

We tested the hypothesis that cancer cachexia progression would induce oxidative post-translational modifications (Ox-PTMs) associated with skeletal muscle wasting, with different responses in muscles with the prevalence of glycolytic and oxidative fibers. We used cysteine-specific isotopic coded affinity tags (OxICAT) and gel-free mass spectrometry analysis to investigate the cysteine Ox-PTMs profile in the proteome of both plantaris (glycolytic) and soleus (oxidative) muscles in tumor-bearing and control rats. Histological analysis revealed muscle atrophy in type II fibers in plantaris muscle, with no changes in plantaris type I fibers and no differences in both soleus type I and II fibers in tumor-bearing rats when compared to healthy controls. Tumor progression altered the Ox-PTMs profile in both plantaris and soleus. However, pathway analysis including the differentially oxidized proteins revealed tricarboxylic acid cycle and oxidative phosphorylation as main affected pathways in plantaris muscle from tumor-bearing rats, while the same analysis did not show main metabolic pathways affected in the soleus muscle. In addition, cancer progression affected several metabolic parameters such as ATP levels and markers of oxidative stress associated with muscle atrophy in plantaris muscle, but not in soleus. However, isolated soleus from tumor-bearing rats had a reduced force production capacity when compared to controls. These novel findings demonstrate that tumor-bearing rats have severe muscle atrophy exclusively in glycolytic fibers. Cancer progression is associated with cysteine Ox-PTMs in the skeletal muscle, but these modifications affect different pathways in a glycolytic muscle compared to an oxidative muscle, indicating that intrinsic muscle oxidative capacity determines the response to cancer cachectic effects.

Objective We tested the hypothesis that exercise training would attenuate metabolic impairment in a model of severe cancer cachexia. Methods We used multiple in vivo and in vitro methods to explore the mechanisms underlying the beneficial effects induced by exercise training in tumor-bearing rats. Results Exercise training improved running capacity, prolonged lifespan, reduced oxidative stress, and normalized muscle mass and contractile function in tumor-bearing rats. An unbiased proteomic screening revealed COP9 signalosome complex subunit 2 (COPS2) as one of the most downregulated proteins in skeletal muscle at the early stage of cancer cachexia. Exercise training normalized muscle COPS2 protein expression in tumor-bearing rats and mice. Lung cancer patients with low endurance capacity had low muscle COPS2 protein expression as compared to age-matched control subjects. To test whether decrease in COPS2 protein levels could aggravate or be an intrinsic compensatory mechanism to protect myotubes from cancer effects, we performed experiments in vitro using primary myotubes. COPS2 knockdown in human myotubes affected multiple cellular pathways, including regulation of actin cytoskeleton. Incubation of cancer-conditioned media in mouse myotubes decreased F-actin expression, which was partially restored by COPS2 knockdown. Direct repeat 4 (DR4) response elements have been shown to positively regulate gene expression. COPS2 overexpression decreased the DR4 activity in mouse myoblasts, and COPS2 knockdown inhibited the effects of cancer-conditioned media on DR4 activity. Conclusions These studies demonstrated that exercise training may be an important adjuvant therapy to counteract cancer cachexia and uncovered novel mechanisms involving COPS2 to regulate myotube homeostasis in cancer cachexia., Highlights • Exercise training prolongs lifespan, reduces oxidative stress, and improves skeletal muscle function in tumor-bearing rats. • Muscle COP9 signalosome complex subunit 2 (COPS2) protein is downregulated in cancer cachexia. • Exercise training restores COPS2 protein expression to control levels. • Cancer-conditioned media decreased F-actin expression in myotubes, which is partially restored by COPS2 knockdown. • COPS2 overexpression decreases DR4 activity and COPS2 knockdown inhibits cancer-conditioned media effects on DR4 activity.

Aims We aimed to investigate the impact of cancer cachexia and previous aerobic exercise training (AET) on cardiac function and structure in tumor bearing mice. Main methods Colon adenocarcinoma cells 26 (CT26) were subcutaneously injected in BALB/c mice to establish robust cancer cachexia model. AET was performed on a treadmill during 45 days, 60 min/5 days per week. Cardiac function was evaluated by echocardiography and cardiac morphology was assessed by light microscopy. The protein expression levels of mitochondrial complex were analyzed by Western blotting. The mRNA levels of genes related to cardiac remodeling and autophagy were analyzed by quantitative Real-Time PCR. Key findings Our data confirms CT26 tumor bearing mice as a well-characterized and robust model of cancer cachexia. CT26 mice exhibited cardiac remodeling and dysfunction characterized by cardiac atrophy and impaired left ventricle ejection fraction paralleled by cardiac necrosis, inflammation and fibrosis. AET partially reversed the left ventricle ejection fraction and led to significant anti-cardiac remodeling effect associated reduced necrosis, inflammation and cardiac collagen deposition in CT26 mice. Reduced TGF-β1 mRNA levels, increased mitochondrial complex IV protein levels and partial recovery of BNIP3 mRNA levels in cardiac tissue were associated with the cardiac effects of AET in CT26 mice. Thus, we suggest AET as a powerful regulator of key pathways involved in cardiac tissue homeostasis in cancer cachexia. Significance Our study provides a robust model of cancer cachexia, as well as highlights the potential and integrative effects of AET as a preventive strategy for reducing cardiac damage in cancer cachexia.

Background Disruption of endoplasmic reticulum (ER) homeostasis is a common feature of cardiac diseases. However, the signaling events involved in ER stress-induced cardiac dysfunction are still elusive. Here, we uncovered a mechanism by which disruption of ER homeostasis impairs cardiac contractility. Methods/results We found that ER stress is associated with activation of JNK and upregulation of BNIP3 in a post-myocardial infarction (MI) model of cardiac dysfunction. Of interest, 4-week treatment of MI rats with the chemical ER chaperone 4-phenylbutyrate (4PBA) prevented both activation of JNK and upregulation of BNIP3, and improved cardiac contractility. We showed that disruption of ER homeostasis by treating adult rat cardiomyocytes in culture with tunicamycin leads to contractile dysfunction through JNK signaling pathway. Upon ER stress JNK upregulates BNIP3 in a FOXO3a-dependent manner. Further supporting a BNIP3 mechanism for ER stress-induced deterioration of cardiac function, siRNA-mediated BNIP3 knockdown mitigated ER stress-induced cardiomyocyte dysfunction by reestablishing sarcoplasmic reticulum Ca2+ content. Conclusions Collectively, our data identify JNK-dependent upregulation of BNIP3 as a critical process involved in ER stress-induced cardiomyocyte contractile dysfunction and highlight 4PBA as a potential intervention to counteract ER stress-mediated BNIP3 upregulation in failing hearts.

Os efeitos benéficos do exercício físico aeróbico (EFA) frente à prevenção e ao tratamento de doenças ocorrem por meio de adaptações agudas e crônicas em diversos tecidos e sistemas orgânicos, dentre os quais o tecido muscular esquelético assume papel de destaque. Considerando que a musculatura esquelética é rica em mitocôndrias, não surpreende o fato de que o aumento da capacidade e potência aeróbicas induzido pelo EFA ocorra principalmente em função das adaptações mitocondriais. Contudo, os mecanismos moleculares envolvidos nessas adaptações mitocondriais induzidas pelo EFA não são completamente compreendidos. Considerando-se que a atividade nervosa simpática aumenta consideravelmente durante a realização de uma sessão de EFA, parte das respostas mitocondriais ao EFA poderia ser mediada pela ativação dos receptores \'beta\'-adrenérgicos (\'beta\'-AR) na musculatura esquelética, representados predominantemente pelo subtipo \'beta\'2 (~ 90%). Com isso, na primeira parte do presente estudo, testou-se a hipótese de que a sinalização \'beta\'2-AR contribuiria para as respostas de função e dinâmica mitocondriais induzidas pela ativação adrenérgica aguda na musculatura esquelética. Para isso, o agonista \'beta\'-AR não seletivo isoproterenol foi administrado na presença ou ausência do antagonista \'beta\'2-AR específico, ICI 118,551, tanto em miotúbulos diferenciados a partir da linhagem de células C2C12 como em camundongos da cepa FVB. Os dados demonstraram que a ativação \'beta\'-AR aumentou significativamente a respiração mitocondrial em miotúbulos e em fibras musculares isoladas do músculo tibial anterior de camundongos, e que tal resposta foi dependente da ativação dos receptores \'beta\'2-AR via eixo proteína Gαs-AMPc-PKA. Em relação à dinâmica mitocondrial, os dados obtidos demonstraram que a ativação dos receptores \'beta\'2-AR aumentou a fusão mitocondrial na musculatura esquelética, e que esta resposta estava associada a um aumento da proteína de fusão Mfn1. Na segunda parte do presente estudo, testou-se a hipótese de que a ativação \'beta\'2-AR contribuiria para as respostas mitocondriais agudas induzidas pelo EFA na musculatura esquelética. Para isso, camundongos FVB foram submetidos a uma sessão de EFA em esteira rolante com e sem o bloqueio dos receptores \'beta\'2-AR com o antagonista ICI 118,551, e parâmetros de função e dinâmica mitocondriais foram avaliados. O EFA foi capaz de aumentar a respiração máxima mitocondrial em fibras musculares isoladas, e este efeito foi parcialmente prevenido pelo bloqueio dos receptores \'beta\'2-AR. Além disso, assim como observado com o tratamento com isoproterenol, o EFA promoveu aumento da fusão mitocondrial na musculatura esquelética, sendo que a ativação \'beta\'2-AR estava associada a essa resposta, porém em uma menor magnitude em relação aos efeitos observados com o estímulo farmacológico. Assim como observado no tratamento com isoproterenol, a expressão de Mfn1 foi aumentada pela sessão de EFA, a qual não foi atenuada pelo bloqueio dos receptores \'beta\'2-AR com ICI 118,551. Dessa forma, a partir dos resultados obtidos no presente estudo, pode-se concluir que os receptores \'beta\'2-AR têm papel importante no metabolismo muscular esquelético, modulando a função e a dinâmica mitocondriais. Essa resposta é evidente mediante ativação direta via agonistas \'beta\'-AR no músculo esquelético, no entanto, os receptores \'beta\'2-AR modulam parcialmente as respostas mitocondriais em resposta à uma sessão de EFA The beneficial effects of aerobic exercise (AE) for prevention and treatment of diseases are due to acute and chronic adaptations in several organ systems, including skeletal muscle, one of the main tissues involved in these adaptations. Since skeletal muscle contains high amounts of mitochondria, the increased aerobic capacity induced by AE occurs mainly due to mitochondrial adaptations. However, the molecular mechanisms underlying skeletal muscle mitochondrial adaptations induced by AE are not fully understood. One of the mechanisms by which these mitochondrial adaptations may occur is sympathetic activation mediated by \'beta\'2-adrenergic receptors (\'beta\'-AR) in skeletal muscle, predominantly represented by \'beta\'2 subtype (~90%). Considering that sympathetic activity increases during AE, \'beta\'2-AR signaling might be involved in skeletal muscle mitochondrial adaptations in response to AE. Thus, in the first part of this study, we tested the hypothesis that acute \'beta\'2-AR activation contributes to mitochondrial function and dynamics adaptations in skeletal muscle. For this, both differentiated C2C12 myotubes and FVB mice were treated with the nonselective \'beta\'-AR agonist isoproterenol in the presence or absence of the specific \'beta\'2-AR antagonist, ICI 118,551. The data showed that β-AR activation significantly increased mitochondrial respiration in myotubes and muscle fiber bundles isolated from mice tibialis anterior, and that this response was dependent on \'beta\'2-AR receptors activation via protein G\' alpha\'s-cAMP-PKA signaling cascade. Regarding mitochondrial dynamics, the data showed that \'beta\'2-AR receptor activation increased mitochondrial fusion in skeletal muscle and that this response was associated to an increase in Mfn1. In the second part of this study, we tested the hypothesis that \'beta\'2-AR activation would contribute to acute mitochondrial responses induced by AE in skeletal muscle. For this, FVB mice underwent one session of AE on a treadmill with and without \'beta\'2-AR receptor blockade with ICI 118,551, and parameters of mitochondrial function and dynamics were evaluated. AE was able to increase maximal mitochondrial respiration in isolated muscle fibers, and this effect was partially prevented by blocking \'beta\'2-AR receptors. Furthermore, as observed with isoproterenol treatment, AE increased mitochondrial fusion in skeletal muscle. \'beta\'2-AR activation was associated with this response, but in a smaller magnitude compared to the effects observed with pharmacological stimulation. As observed with the isoproterenol treatment, Mfn1 expression was increased by AE, which was not attenuated by blocking \'beta\'2-AR receptors with ICI 118,551. Therefore, one can conclude that \'beta\'2-AR receptors play an important role in skeletal muscle metabolism, modulating mitochondrial function and dynamics. This response is evident by direct activation through \'beta\'-AR agonists in skeletal muscle, however, \'beta\'2-AR receptors partially modulate mitochondrial responses induced by AE

Increased proteolytic activity has been widely associated with skeletal muscle atrophy. However, elevated proteolysis is also critical for the maintenance of cellular homeostasis by disposing cytotoxic proteins and non-functioning organelles. We recently demonstrated that exercise activates autophagy and re-establishes proteostasis in cardiac diseases. Here, we characterized the impact of exercise on skeletal muscle autophagy and proteostasis in a model of neurogenic myopathy induced by sciatic nerve constriction in rats. Neurogenic myopathy, characterized by progressive atrophy and impaired contractility, was paralleled by accumulation of autophagy-related markers and loss of acute responsiveness to both colchicine and chloroquine. These changes were correlated with elevated levels of damaged proteins, chaperones and pro-apoptotic markers compared to control animals. Sustained autophagy inhibition using chloroquine in rats (50 mg.kg−1.day−1) or muscle-specific deletion of Atg7 in mice was sufficient to impair muscle contractility in control but not in neurogenic myopathy, suggesting that dysfunctional autophagy is critical in skeletal muscle pathophysiology. Finally, 4 weeks of aerobic exercise training (moderate treadmill running, 5x/week, 1 h/day) prior to neurogenic myopathy improved skeletal muscle autophagic flux and proteostasis. These changes were followed by spared muscle mass and better contractility properties. Taken together, our findings suggest the potential value of exercise in maintaining skeletal muscle proteostasis and slowing down the progression of neurogenic myopathy. pen Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

O objetivo deste artigo de revisão sistemática foi apresentar o envolvimento da sinalização de aminoácidos e mecanotransdução na ativação do complexo 1 do alvo da rapamicina em mamíferos (mTORC1) na musculatura esquelética de animais e a expressão e papel do mTORC1 em humanos submetidos ao treinamento de força/estímulo mecânico. Foi realizada uma busca na base de dados PubMed com as seguintes palavras-chave: mTORC1, mammalian target of rapamycin complex 1, resistance exercise, strength training, mechanical overload e skeletal muscle hypertrophy, amino acid sensing transporter e mechanotransduction. Evidências demonstram que a ativação do mTORC1 possui correlação positiva com a hipertrofia muscular induzida pelo treinamento de força/estímulo mecânico. O mTORC1 integra diversos sinais oriundos de aminoácidos (sinalização de transportadores e sensores) e estímulo mecânico/treinamento de força (mecanotransdução). Ademais, o emprego de modelos de camundongos mutantes, abordagens genéticas, farmacológicas, cultura de células, modelos experimentais de treinamento de força para animais, assim como estudos com humanos, vêm possibilitando a elucidação destes mecanismos moleculares.

Background Skeletal muscle wasting is associated with poor prognosis and increased mortality in heart failure (HF) patients. Glycolytic muscles are more susceptible to catabolic wasting than oxidative ones. This is particularly important in HF since glycolytic muscle wasting is associated with increased levels of reactive oxygen species (ROS). However, the main ROS sources involved in muscle redox imbalance in HF have not been characterized. Therefore, we hypothesized that NADPH oxidases would be hyperactivated in the plantaris muscle of infarcted rats, contributing to oxidative stress and hyperactivation of the ubiquitin-proteasome system (UPS), ultimately leading to atrophy. Methods Rats were submitted to myocardial infarction (MI) or Sham surgery. Four weeks after surgery, MI and Sham groups underwent eight weeks of treatment with apocynin, a NADPH oxidase inhibitor, or placebo. NADPH oxidase activity, oxidative stress markers, NF-κB activity, p38 MAPK phosphorylation, mRNA and sarcolemmal protein levels of NADPH oxidase components, UPS activation and fiber cross-sectional area were assessed in the plantaris muscle. Results The plantaris of MI rats displayed atrophy associated with increased Nox2 mRNA and sarcolemmal protein levels, NADPH oxidase activity, ROS production, lipid hydroperoxides levels, NF-κB activity, p38 MAPK phosphorylation and UPS activation. NADPH oxidase inhibition by apocynin prevented MI-induced skeletal muscle atrophy by reducing ROS production, NF-κB hyperactivation, p38 MAPK phosphorylation and proteasomal hyperactivity. Conclusion Our data provide evidence for NADPH oxidase hyperactivation as an important source of ROS production leading to plantaris atrophy in heart failure rats, suggesting that this enzyme complex plays key role in skeletal muscle wasting in HF.

The current study tested the hypotheses that 1) an acute bout of aerobic exercise impairs isolated skeletal muscle contractile properties and 2) N-acetylcysteine (a thiol antioxidant; NAC) administration can restore the impaired muscle contractility after exercise.At rest or immediately after an acute bout of aerobic exercise, extensor digitorum longus (EDL) and soleus muscles from male Wistar rats were harvested for ex vivo skeletal muscle contraction experiments. Muscles from exercised animals were incubated in Krebs Ringer's buffer in absence or presence of 20mM of NAC. Force capacity and fatigue properties were evaluated.Exercised EDL and soleus displayed lower force production across various stimulation frequencies (p0.001), indicating that skeletal muscle force production was impaired after an acute bout of exercise. However, NAC treatment restored the loss of force production in both EDL and soleus after fatiguing exercise (p0.05). Additionally, NAC treatment increased relative force production at different time points during a fatigue-induced protocol, suggesting that NAC treatment mitigates fatigue induced by successive contractions.NAC treatment improves force capacity and fatigue properties in ex vivo skeletal muscle from rats submitted to an acute bout of aerobic exercise.

β(2)-adrenergic receptor (β(2)-AR) agonists have been used as ergogenics by athletes involved in training for strength and power in order to increase the muscle mass. Even though anabolic effects of β(2)-AR activation are highly recognized, less is known about the impact of β(2)-AR in endurance capacity. We presently used mice lacking β(2)-AR [β(2)-knockout (β(2) KO)] to investigate the role of β(2)-AR on exercise capacity and skeletal muscle metabolism and phenotype. β(2) KO mice and their wild-type controls (WT) were studied. Exercise tolerance, skeletal muscle fiber typing, capillary-to-fiber ratio, citrate synthase activity and glycogen content were evaluated. When compared with WT, β(2) KO mice displayed increased exercise capacity (61%) associated with higher percentage of oxidative fibers (21% and 129% of increase in soleus and plantaris muscles, respectively) and capillarity (31% and 20% of increase in soleus and plantaris muscles, respectively). In addition, β(2) KO mice presented increased skeletal muscle citrate synthase activity (10%) and succinate dehydrogenase staining. Likewise, glycogen content (53%) and periodic acid-Schiff staining (glycogen staining) were also increased in β(2) KO skeletal muscle. Altogether, these data provide evidence that disruption of β(2)-AR improves oxidative metabolism in skeletal muscle of β(2) KO mice and this is associated with increased exercise capacity.

We have recently demonstrated that NADPH oxidase hyperactivity, NF-κB activation, and increased p38 phosphorylation lead to atrophy of glycolytic muscle in heart failure (HF). Aerobic exercise training (AET) is an efficient strategy to counteract skeletal muscle atrophy in this syndrome. Therefore, we tested whether AET would regulate muscle redox balance and protein degradation by decreasing NADPH oxidase hyperactivity and reestablishing NF-κB signaling, p38 phosphorylation, and proteasome activity in plantaris muscle of myocardial infarcted-induced HF (MI) rats. Thirty-two male Wistar rats underwent MI or fictitious surgery (SHAM) and were randomly assigned into untrained (UNT) and trained (T; 8 wk of AET on treadmill) groups. AET prevented HF signals and skeletal muscle atrophy in MI-T, which showed an improved exercise tolerance, attenuated cardiac dysfunction and increased plantaris fiber cross-sectional area. To verify the role of inflammation and redox imbalance in triggering protein degradation, circulating TNF-α levels, NADPH oxidase profile, NF-κB signaling, p38 protein levels, and proteasome activity were assessed. MI-T showed a reduced TNF-α levels, NADPH oxidase activity, and Nox2 mRNA expression toward SHAM-UNT levels. The rescue of NADPH oxidase activity induced by AET in MI rats was paralleled by reducing nuclear binding activity of the NF-κB, p38 phosphorylation, atrogin-1, mRNA levels, and 26S chymotrypsin-like proteasome activity. Taken together our data provide evidence for AET improving plantaris redox homeostasis in HF associated with a decreased NADPH oxidase, redox-sensitive proteins activation, and proteasome hyperactivity further preventing atrophy. These data reinforce the role of AET as an efficient therapy for muscle wasting in HF. NEW & NOTEWORTHY This study demonstrates, for the first time, the contribution of aerobic exercise training (AET) in decreasing muscle NADPH oxidase activity associated with reduced reactive oxygen species production and systemic inflammation, which diminish NF-κB overactivation, p38 phosphorylation, and ubiquitin proteasome system hyperactivity. These molecular changes counteract plantaris atrophy in trained myocardial infarction-induced heart failure rats. Our data provide new evidence into how AET may regulate protein degradation and thus prevent skeletal muscle atrophy.

Cardiac endoplasmic reticulum (ER) stress through accumulation of misfolded proteins plays a pivotal role in cardiovascular diseases. In an attempt to reestablish ER homoeostasis, the unfolded protein response (UPR) is activated. However, if ER stress persists, sustained UPR activation leads to apoptosis. There is no available therapy for ER stress relief. Considering that aerobic exercise training (AET) attenuates oxidative stress, mitochondrial dysfunction and calcium imbalance, it may be a potential strategy to reestablish cardiac ER homoeostasis. We test the hypothesis that AET would attenuate impaired cardiac ER stress after myocardial infarction (MI). Wistar rats underwent to either MI or sham surgeries. Four weeks later, rats underwent to 8 weeks of moderate‐intensity AET. Myocardial infarction rats displayed cardiac dysfunction and lung oedema, suggesting heart failure. Cardiac dysfunction in MI rats was paralleled by increased protein levels of UPR markers (GRP78, DERLIN‐1 and CHOP), accumulation of misfolded and polyubiquitinated proteins, and reduced chymotrypsin‐like proteasome activity. These results suggest an impaired cardiac protein quality control. Aerobic exercise training improved exercise capacity and cardiac function of MI animals. Interestingly, AET blunted MI‐induced ER stress by reducing protein levels of UPR markers, and accumulation of both misfolded and polyubiquinated proteins, which was associated with restored proteasome activity. Taken together, our study provide evidence for AET attenuation of ER stress through the reestablishment of cardiac protein quality control, which contributes to better cardiac function in post‐MI heart failure rats. These results reinforce the importance of AET as primary non‐pharmacological therapy to cardiovascular disease.

A insuficiência cardíaca (IC) é uma síndrome complexa que envolve múltiplos sistemas e mecanismos compensatórios neuro-hormonais, acompanhada de altos índices de morbidade e mortalidade, e caracterizada por sintomas clínicos como fadiga, dispneia, e intolerância aos esforços físicos. Embora a IC seja uma síndrome de origem cardíaca, observam-se alterações em outros tecidos, como na musculatura esquelética. As modificações do fenótipo muscular e a perda de massa muscular esquelética observadas na IC contribuem para o mau prognóstico e para o aumento da mortalidade dos pacientes. Considerando que os receptores 2-adrenérgicos medeiam a atividade do sistema nervoso simpático na musculatura esquelética, e que a hiperatividade simpática é um dos principais componentes envolvidos no desenvolvimento da miopatia esquelética observada IC, sugere-se que estes receptores estejam associados às alterações morfofuncionais da musculatura esquelética na síndrome. Na presente Dissertação, avaliamos a contribuição dos receptores 2-adrenérgicos nas alterações metabólicas e morfofuncionais da musculatura esquelética e na intolerância aos esforços físicos decorrentes da IC. Para isso, utilizamos camundongos da linhagem FVB controles e com inativação gênica dos receptores 2-adrenérgicos (2KO) que foram submetidos à cirurgia de infarto ou à cirurgia fictícia (sham). O infarto induziu disfunção e remodelamento cardíacos nos animais controles, acompanhados de aumento nos níveis de noradrenalina e adrenalina circulantes, intolerância ao esforço físico, mudança na tipagem de fibras musculares e rarefação capilar nos músculos sóleo e plantar. Essas mesmas respostas foram observadas nos animais com inativação gênica dos receptores 2-adrenérgicos após o infarto do miocárdio, no entanto, os animais 2KO infartados apresentaram uma maior redução da tolerância ao esforço físico e um quadro mais grave de miopatia esquelética. Acompanhando essa atrofia muscular esquelética presente nos animais 2KO infartados, foi observado aumento da expressão de proteínas relacionadas ao sistema proteolítico ubiquitina-proteassoma, aumento da atividade do 26S-proteassoma e tendência a diminuição na expressão proteica de p-Akt (ser473), p-4E-BP1 (thr37/46) e p-FoxO3 (ser253). Os resultados sugerem que a ausência dos receptores 2-adrenérgicos agrava e/ou antecipa as alterações musculoesqueléticas observadas na insuficiência cardíaca, principalmente a atrofia muscular, e que a ativação do sistema proteolítico ubiquitina proteassoma e a inibição da síntese proteica por meio da via Akt/4E-BP1 parecem colaborar para estas respostas Heart failure (HF) is a complex syndrome involving multiple systems and neurohumoral compensatory mechanisms accompanied by high morbidity and mortality, and it is characterized by clinical signs like fatigue, dyspnea, and increased exercise intolerance. Although HF is a syndrome of cardiac origin, it promotes changes in other tissues, such as skeletal muscle, where modifications of muscle phenotype and the loss of skeletal muscle mass observed in HF contribute to poor prognosis and increased mortality of patients. Taking into consideration that 2-adrenoceptors mediate the activity of sympathetic nervous system in skeletal muscle and that sympathetic hyperactivity is one of the main components involved in developing skeletal muscle abnormalities in HF, it has been suggested that 2-adrenoceptors would be associated with morphofunctional alterations due to chronic sympathetic hyperactivity in skeletal muscles in HF. In the present study, we evaluated the contribution of 2-adrenoceptors on metabolic and morphofunctional alterations in skeletal muscle and also on exercise intolerance-induced by HF. For this, we used FVB mice and mice lacking 2-adrenoceptors (2KO) that were submitted to myocardial infarction or sham surgery. Myocardial infarction induced cardiac dysfunction and remodeling in FVB mice, which was accompanied by significantly increased plasma levels of norepinephrine and epinephrine, exercise intolerance, changes in muscle fiber type and vascular rarefaction in both soleus and plantaris muscles. These same responses were observed in mice lacking 2-adrenoceptors submitted to myocardial infarction. However, infarcted 2KO mice presented a higher decrease of exercise tolerance and more severe skeletal myopathy. Accompanying the skeletal muscle atrophy of infarcted 2KO mice, it was observed a significantly increased protein expression of proteins related with the ubiquitin-proteasome system, an increased 26S-proteassome activity and a trend toward decreased protein expression of p-Akt (ser473), p-4E-BP1 (thr37/46) and p- FoxO3a (ser253). These results suggest that the lack of 2-adrenoceptors worsens and/or anticipates the skeletal muscle alterations observed in HF, mainly muscular atrophy, and the activation of ubiquitin-proteassome system and inhibition of protein synthesis by Akt/4E-BP1 pathway seem to play an important role in skeletal muscle myopathy

Skeletal myopathy has been identified as a major comorbidity of heart failure (HF) affecting up to 20% of ambulatory patients leading to shortness of breath, early fatigue, and exercise intolerance. Neurohumoral blockade, through the inhibition of renin angiotensin aldosterone system (RAS) andβ-adrenergic receptor blockade (β-blockers), is a mandatory pharmacological therapy of HF since it reduces symptoms, mortality, and sudden death. However, the effect of these drugs on skeletal myopathy needs to be clarified, since exercise intolerance remains in HF patients optimized withβ-blockers and inhibitors of RAS. Aerobic exercise training (AET) is efficient in counteracting skeletal myopathy and in improving functional capacity and quality of life. Indeed, AET has beneficial effects on failing heart itself despite being of less magnitude compared with neurohumoral blockade. In this way, AET should be implemented in the care standards, together with pharmacological therapies. Since both neurohumoral inhibition and AET have a direct and/or indirect impact on skeletal muscle, this review aims to provide an overview of the isolated effects of these therapeutic approaches in counteracting skeletal myopathy in HF. The similarities and dissimilarities of neurohumoral inhibition and AET therapies are also discussed to identify potential advantageous effects of these combined therapies for treating HF.

This study aimed to investigate mitophagy-related signals in skeletal muscle following a single bout of aerobic exercise in mice. C57BL6 mice were submitted to a single bout of treadmill aerobic exercise for 80 minutes at 60% of maximal speed achieved in a graded treadmill test. Skeletal muscles were harvested at rest, immediately after the bout of aerobic exercise and after 3, 6, 12 and 24h of recovery. One hindlimb was used for fresh isolation of skeletal muscle mitochondrial-enriched fraction (MF), and contralateral muscles were immediately frozen. Immunoblotting for LC3, p62 and Bnip3 were performed either in quadriceps muscles or MF. qPCR were performed to evaluate mRNA levels of LC3, p62 and Bnip3 in quadriceps muscles. No protein was differentially expressed in total lysate of quadriceps muscle. However, LC3-II levels were significantly increased in MF immediately after the bout of aerobic exercise (72.5%) and after 12h of recovery (63.3%), whereas p62 protein levels were increased at 6 (30%) and 1...

The beneficial effects of physical exercise for health are due to physiological adaptations that coordinate several organs. In short term, these adaptations occur to supply the increased metabolic demand imposed by exercising muscles, and in long term, to reduce the homeostatic disturbance caused by exercise training. In this sense, the autonomic nervous system plays a crucial role in integrating these short- and long-term physical exercise adjustments by modulating the sympathetic and parasympathetic outflows in health and disease conditions. Additionally, accumulated evidences have shown that exercise training is an efficient strategy for treatment and prevention of cardiovascular diseases. For instance, one striking effect of exercise training is a reduction in sympathetic hyperactivity observed in heart failure. This response results in a better autonomic control of cardiovascular system by improving the cardiac and vascular adrenergic responses to exercise stimulus in heart failure.

Skeletal myopathy is a hallmark of heart failure (HF) and has been associated with a poor prognosis. HF and other chronic degenerative diseases share a common feature of a stressed system: sympathetic hyperactivity. Although beneficial acutely, chronic sympathetic hyperactivity is one of the main triggers of skeletal myopathy in HF. Considering that β2 -adrenoceptors mediate the activity of sympathetic nervous system in skeletal muscle, we presently evaluated the contribution of β2 -adrenoceptors for the morphofunctional alterations in skeletal muscle and also for exercise intolerance induced by HF. Male WT and β2 -adrenoceptor knockout mice on a FVB genetic background (β2 KO) were submitted to myocardial infarction (MI) or SHAM surgery. Ninety days after MI both WT and β2 KO mice presented to cardiac dysfunction and remodelling accompanied by significantly increased norepinephrine and epinephrine plasma levels, exercise intolerance, changes towards more glycolytic fibres and vascular rarefaction in plantaris muscle. However, β2 KO MI mice displayed more pronounced exercise intolerance and skeletal myopathy when compared to WT MI mice. Skeletal muscle atrophy of infarcted β2 KO mice was paralleled by reduced levels of phosphorylated Akt at Ser 473 while increased levels of proteins related with the ubiquitin--proteasome system, and increased 26S proteasome activity. Taken together, our results suggest that lack of β2 -adrenoceptors worsen and/or anticipate the skeletal myopathy observed in HF.

Made available in DSpace on 2014-09-30T18:18:31Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-06-01Bitstream added on 2014-10-01T14:03:24Z : No. of bitstreams: 1 S1517-86922013000300012.pdf: 259455 bytes, checksum: 30704e3d729ec41395df6f17b108afb4 (MD5) INTRODUÇÃO: São escassos os estudos os quais verificaram os efeitos do exercício físico sobre os biomarcadores de estresse oxidativo e sua relação com o glicogênio muscular. OBJETIVO: O presente estudo verificou o efeito do treinamento aeróbio de natação (TAN) sobre os biomarcadores de estresse oxidativo, conteúdo de glicogênio e crescimento celular no músculo esquelético de ratos. MÉTODOS: Dezoito ratos Wistar macho (60 dias) foram divididos em dois grupos: Grupo Sedentário (SG; n = 10): ratos sedentários; e Grupo Treinado (TG; n = 8): ratos submetidos ao TAN (5,0 % do peso corporal), 1h/dia, 5X/semana, durante oito semanas. A atividade das enzimas antioxidantes (AEA) superóxido dismutase (SOD; U/ml), catalase (CAT; µmol/min/100 mg) e glutationa peroxidase (GPx; nmol/min/100 mg), bem como a concentração das substâncias que reagem ao ácido tiobarbitúrico (TBARs; nmolMDA/mg proteína) foram determinadas no músculo gastrocnêmio direito. Os conteúdos de glicogênio (mg/100 mg), proteína (g/100 g) e DNA (g/100 g) foram mensurados no músculo gastrocnêmio esquerdo. RESULTADOS: A AEA foi maior no TG (CAT: 0,87 ± 0,04; SOD: 6,49 ± 0,045; GPX: 6,49 ± 0,52) quando comparados com SG (CAT: 0,52 ± 0,03; SOD: 4,1 ± 0,37; GPx: 2,94 ± 0,56). Os níveis de TBARs foram menores em TG (TG: 2,35 ± 0,41; SG: 8,90 ± 0,47). O conteúdo de glicogênio muscular (SG: 0,108 ± 0,013; TG: 0,320 ± 0,012) e a razão proteína/DNA (SG: 24,94 ± 3,25; TG: 41,68 ± 4,02) foram maiores no TG. CONCLUSÃO: Em conjunto, estes dados confirmam que o TAN melhorou a defesa antioxidante, a qual pode estar associada ao aumento do conteúdo de glicogênio no músculo esquelético dos animais. INTRODUCTION: Studies which verified the effects of physical exercise on oxidative stress biomarkers and its relation to muscle glycogen are lack. OBJECTIVE: The present study verified the effects of aerobic swimming training (AST) on biomarkers of oxidative stress, glycogen content and cell growth in the skeletal muscle of rats. METHODS: Eighteen male Wistar rats (60 days) were divided into two groups: Sedentary Group (SG; n = 10): sedentary rats; and Trained Group (TG; n = 8): rats subjected to AST (5.0% of body weight), 1h/day, 5x/week, during 8 weeks. The activity of antioxidant enzymes (AOE) superoxide dismutase (SOD; U/ml), catalase (CAT; µmol/min/100mg), glutathione peroxidase (GPx; nmol/min/100mg), and substances that react with thiobarbituric acid levels (TBARs; nmolMDA/mg protein) were determined in the right gastrocnemius muscle. Glycogen (mg/100mg), protein (g/100g), and DNA (g/100g) contents were evaluated in the left gastrocnemius muscle. Data were analyzed by Student t-test (p < 0.05). RESULTS: AOE activity was higher in the TG group (CAT: 0.87 ± 0.04; SOD: 6.49 ± 0.45; GPX: 6.49 ± 0.52) when compared to SG group (CAT: 0.52 ± 0.03; SOD: 4.10 ± 0.37; GPx: 2.87 ± 0.35). TBARs levels was lower in TG (TG: 2.35 ± 0.45; SG: 8.90 ± 0.47). Gastrocnemius glycogen content (SG: 0.108 ± 0.013; TG: 0.320 ± 0.012) and protein/DNA ratio (SG: 24.94 ± 3.25; TG: 41.68 ± 4.02) were higher in TG group. CONCLUSION: Altogether, these data provide evidence that AST improved antioxidant defense, which may be associated to higher glycogen content of skeletal muscle of the animals. UFMT Programa de Pós-Graduação em Biociências/Nutrição USP Escola de Educação Física e Esporte UFMT Programa de Pós-Graduação em Educação Física UNIVAG Universidade Estadual Paulista Universidade Estadual Paulista

INTRODUÇÃO: São escassos os estudos os quais verificaram os efeitos do exercício físico sobre os biomarcadores de estresse oxidativo e sua relação com o glicogênio muscular. OBJETIVO: O presente estudo verificou o efeito do treinamento aeróbio de natação (TAN) sobre os biomarcadores de estresse oxidativo, conteúdo de glicogênio e crescimento celular no músculo esquelético de ratos. MÉTODOS: Dezoito ratos Wistar macho (60 dias) foram divididos em dois grupos: Grupo Sedentário (SG; n = 10): ratos sedentários; e Grupo Treinado (TG; n = 8): ratos submetidos ao TAN (5,0 % do peso corporal), 1h/dia, 5X/semana, durante oito semanas. A atividade das enzimas antioxidantes (AEA) superóxido dismutase (SOD; U/ml), catalase (CAT; µmol/min/100 mg) e glutationa peroxidase (GPx; nmol/min/100 mg), bem como a concentração das substâncias que reagem ao ácido tiobarbitúrico (TBARs; nmolMDA/mg proteína) foram determinadas no músculo gastrocnêmio direito. Os conteúdos de glicogênio (mg/100 mg), proteína (g/100 g) e DNA (g/100 g) foram mensurados no músculo gastrocnêmio esquerdo. RESULTADOS: A AEA foi maior no TG (CAT: 0,87 ± 0,04; SOD: 6,49 ± 0,045; GPX: 6,49 ± 0,52) quando comparados com SG (CAT: 0,52 ± 0,03; SOD: 4,1 ± 0,37; GPx: 2,94 ± 0,56). Os níveis de TBARs foram menores em TG (TG: 2,35 ± 0,41; SG: 8,90 ± 0,47). O conteúdo de glicogênio muscular (SG: 0,108 ± 0,013; TG: 0,320 ± 0,012) e a razão proteína/DNA (SG: 24,94 ± 3,25; TG: 41,68 ± 4,02) foram maiores no TG. CONCLUSÃO: Em conjunto, estes dados confirmam que o TAN melhorou a defesa antioxidante, a qual pode estar associada ao aumento do conteúdo de glicogênio no músculo esquelético dos animais. INTRODUCTION: Studies which verified the effects of physical exercise on oxidative stress biomarkers and its relation to muscle glycogen are lack. OBJECTIVE: The present study verified the effects of aerobic swimming training (AST) on biomarkers of oxidative stress, glycogen content and cell growth in the skeletal muscle of rats. METHODS: Eighteen male Wistar rats (60 days) were divided into two groups: Sedentary Group (SG; n = 10): sedentary rats; and Trained Group (TG; n = 8): rats subjected to AST (5.0% of body weight), 1h/day, 5x/week, during 8 weeks. The activity of antioxidant enzymes (AOE) superoxide dismutase (SOD; U/ml), catalase (CAT; µmol/min/100mg), glutathione peroxidase (GPx; nmol/min/100mg), and substances that react with thiobarbituric acid levels (TBARs; nmolMDA/mg protein) were determined in the right gastrocnemius muscle. Glycogen (mg/100mg), protein (g/100g), and DNA (g/100g) contents were evaluated in the left gastrocnemius muscle. Data were analyzed by Student t-test (p < 0.05). RESULTS: AOE activity was higher in the TG group (CAT: 0.87 ± 0.04; SOD: 6.49 ± 0.45; GPX: 6.49 ± 0.52) when compared to SG group (CAT: 0.52 ± 0.03; SOD: 4.10 ± 0.37; GPx: 2.87 ± 0.35). TBARs levels was lower in TG (TG: 2.35 ± 0.45; SG: 8.90 ± 0.47). Gastrocnemius glycogen content (SG: 0.108 ± 0.013; TG: 0.320 ± 0.012) and protein/DNA ratio (SG: 24.94 ± 3.25; TG: 41.68 ± 4.02) were higher in TG group. CONCLUSION: Altogether, these data provide evidence that AST improved antioxidant defense, which may be associated to higher glycogen content of skeletal muscle of the animals.

Aims Increased cardiac sympathetic neuron (SN) activity has been associated with pathologies such as heart failure and hypertrophy, suggesting that cardiac innervation regulates cardiomyocyte trophism. Whether continuous input from the SNs is required for the maintenance of the cardiomyocyte size has not been determined thus far. Methods and results To address the role of cardiac innervation in cardiomyocyte size regulation, we monitored the effect of pharmacological sympathetic denervation in mice on cardiac structure, function, and signalling from 24 h to 30 days in the absence of other pathological stimuli. SN ablation caused an immediate reduction in the cardiomyocyte size with minimal consequences on the resting contractile function. Atrophic remodelling was mediated by the ubiquitin–proteasome system through FOXO-dependent early induction of the muscle-specific E3 ubiquitin ligases Atrogin-1/MAFbx and MuRF1, which was followed by activation of the autophagy–lysosome system. MuRF1 was found to be determinant in denervation atrophy as remodelling did not develop in denervated MuRF1 knock-out (KO) hearts. These effects were caused by decreased basal stimulation of cardiomyocyte β2-adrenoceptor (AR), as atrophy was prevented by treatment of denervated mice with the β2-AR agonist clenbuterol. Consistent with these data, we also observed that β2-AR KO mice showed cardiac atrophy at rest. Conclusion Cardiac SNs are strong regulators of the cardiomyocyte size via β2-AR-dependent repression of proteolysis, demonstrating that the neuro-cardiac axis operates constitutively for the determination of the physiological cardiomyocyte size. These results are of great clinical relevance given the role of β-AR in cardiovascular diseases and their modulation in therapy.

The present investigation was undertaken to test whether exercise training (ET) associated with AMPK/PPAR agonists (EM) would improve skeletal muscle function in mdx mice. These drugs have the potential to improve oxidative metabolism. This is of particular interest because oxidative muscle fibers are less affected in the course of the disease than glycolitic counterparts. Therefore, a cohort of 34 male congenic C57Bl/10J mdx mice included in this study was randomly assigned into four groups: vehicle solution (V), EM [AICAR (AMPK agonist, 50 mg/Kg-1.day-1, ip) and GW 1516 (PPARδ agonist, 2.5 mg/Kg-1.day-1, gavage)], ET (voluntary running on activity wheel) and EM+ET. Functional performance (grip meter and rotarod), aerobic capacity (running test), muscle histopathology, serum creatine kinase (CK), levels of ubiquitined proteins, oxidative metabolism protein expression (AMPK, PPAR, myoglobin and SCD) and intracellular calcium handling (DHPR, SERCA and NCX) protein expression were analyzed. Treatments started when the animals were two months old and were maintained for one month. A significant functional improvement (p