Oral glutamine supplementation relieves muscle loss in immobilized rats, altering p38MAPK and FOXO3a signaling pathways.

BACKGROUND: Skeletal muscle synthesizes, stores, and releases body L-glutamine (GLN). Muscle atrophy due to disabling diseases triggers the activation of proteolytic and pro-apoptotic cell signaling, thus impairing the body's capacity to manage GLN content. This situation has a poor therapeutic prognosis. OBJECTIVE: Evaluating if oral GLN supplementation can attenuate muscle wasting mediated by elevated plasma cortisol and activation of caspase-3, p38MAPK, and FOXO3a signaling pathways in soleus and gastrocnemius muscles of rats submitted to 14-day bilateral hindlimbs immobilization. METHODS: Animals were randomly distributed into six groups: non-immobilized rats (Control), control orally supplemented with GLN (1 g kg-1) in solution with L-alanine (ALA: 0.61 g kg-1; GLN+ALA), control orally supplemented with dipeptide L-alanyl-L-glutamine (DIP; 1.49 g kg-1), hindlimbs immobilized rats (IMOB), IMOB orally GLN+ALA supplemented (GLN+ALA-IMOB), and IMOB orally DIP supplemented (DIP-IMOB). Plasma and muscle GLN concentration, plasma cortisol level, muscle caspase-3 activity, muscle p38MAPK and FOXO3a protein content (total and phosphorylated forms), and muscle cross-sectional area (CSA) were measured. RESULTS: Compared to controls, IMOB rats presented: a) increased plasma cortisol levels; b) decreased plasma and muscle GLN concentration; c) increased muscle caspase-3 activity; d) increased total and phosphorylated p38MAPK protein content; e) increased FOXO3a and decreased phosphorylated FOXO3a protein content; f) reduced muscle weight and CSA befitting to atrophy. Oral supplementation with GLN+ALA and DIP was able to significantly attenuate these effects. CONCLUSIONS: These findings attest that oral GLN supplementation in GLN+ALA solution or DIP forms attenuates rats' skeletal muscle mass wasting caused by disuse-mediated muscle atrophy.

Oral glutamine supplementation attenuates inflammation and oxidative stress-mediated skeletal muscle protein content degradation in immobilized rats: Role of 70 kDa heat shock protein.

Skeletal muscle disuse results in myofibrillar atrophy and protein degradation, via inflammatory and oxidative stress-mediated NF-kB signaling pathway activation. Nutritional interventions, such as l-glutamine (GLN) supplementation have shown antioxidant properties and cytoprotective effects through the modulation on the 70-kDa heat shock protein (HSP70) expression. However, these GLN-mediated effects on cell signaling pathways and biochemical mechanisms that control the myofibrillar protein content degradation in muscle disuse situations are poorly known yet. This study investigated the effects of oral GLN plus l-alanine (ALA; GLN â€‹+ â€‹ALA-solution) supplementation, either in their free or dipeptide (L-alanyl-l-glutamine-DIP) form, on GLN-glutathione (GSH) axis and cytoprotection mediated by HSP70 protein expression in the slow-twitch soleus and fast-twitch gastrocnemius skeletal muscle of rats submitted to 14-days of hindlimb immobilization-induced disuse muscle atrophy. Forty-eight Wistar rats were distributed into 6 groups: hindlimb immobilized (IMOB group) and hindlimb immobilized orally supplemented with either GLN (1 g kg-1) plus ALA (0.61 g kg-1) â€‹(GLN â€‹+ â€‹ALA-IMOB group) or 1.49 â€‹g â€‹kg-1 of DIP (DIP-IMOB group) and; no-immobilized (CTRL) and no-immobilized supplemented GLN â€‹+ â€‹ALA and DIP baselines groups. All animals, including CTRL and IMOB rats (water), were supplemented via intragastric gavage for 14 days, concomitantly to immobilization period. Plasma and muscle GLN levels, lipid (thiobarbituric acid reactive substances-TBARS) and protein (carbonyl) peroxidation, erythrocyte concentration of reduced GSH and GSH disulfide (GSSG), plasma and muscle pro-inflammatory TNF-α levels, muscle IKKα/ß-NF-kB signaling pathway and, the myofibrillar protein content (MPC) were measured. The MPC was significantly lower in IMOB rats, compared to CTRL, GLN â€‹+ â€‹ALA, and DIP animals (p â€‹< â€‹0.05). This finding was associated with reduced plasma and muscle GLN concentration, equally in IMOB animals. Conversely, both GLN â€‹+ â€‹ALA and DIP supplementation restored plasma and muscle GLN levels, which equilibrated GSH and intracellular redox status (GSSG/GSH ratio) in erythrocytes and skeletal muscle even as, increased muscle HSP70 protein expression; attenuating oxidative stress and TNF-α-mediated NF-kB pathway activation, fact that reverberated on reduction of MPC degradation in GLN â€‹+ â€‹ALA-IMOB and DIP-IMOB animals (p â€‹< â€‹0.05). In conclusion, the findings shown herein support the oral GLN â€‹+ â€‹ALA and DIP supplementations as a therapeutic and effective nutritional alternative to attenuate the deleterious effects of the skeletal muscle protein degradation induced by muscle disuse.

L-glutamine supplementations enhance liver glutamine-glutathione axis and heat shock factor-1 expression in endurance-exercise trained rats.

Liver L-glutamine is an important vehicle for the transport of ammonia and intermediary metabolism of amino acids between tissues, particularly under catabolic situations, such as high-intensity exercise. Hence, the aim of this study was to investigate the effects of oral supplementations with L-glutamine in its free or dipeptide forms (with L-alanine) on liver glutamine-glutathione (GSH) axis, and 70 kDa heat shock proteins (HSP70)/heat shock transcription factor 1 (HSF1) expressions. Adult male Wistar rats were 8-week trained (60 min/day, 5 days/week) on a treadmill. During the last 21 days, the animals were daily supplemented with 1 g of L-glutamine/kg body weight per day in either l-alanyl-L-glutamine dipeptide (DIP) form or a solution containing L-glutamine and l-alanine in their free forms (GLN+ALA) or water (controls). Exercise training increased cytosolic and nuclear HSF1 and HSP70 expression, as compared with sedentary animals. However, both DIP and GLN+ALA supplements enhanced HSF1 expression (in both cytosolic and nuclear fractions) in relation to exercised controls. Interestingly, HSF1 rises were not followed by enhanced HSP70 expression. DIP and GLN+ALA supplements increased plasma glutamine concentrations (by 62% and 59%, respectively) and glutamine to glutamate plasma ratio in relation to trained controls. This was in parallel with a decrease in plasma ammonium levels. Supplementations increased liver GSH (by 90%), attenuating the glutathione disulfide (GSSG) to GSH ratio, suggesting a redox state protection. In conclusion, oral administration with DIP and GLN+ALA supplements in endurance-trained rats improve liver glutamine-GSH axis and modulate HSF1 pathway.

Alanyl-glutamine and glutamine plus alanine supplements improve skeletal redox status in trained rats: involvement of heat shock protein pathways.

AIMS: We hypothesized that oral l-glutamine supplementations could attenuate muscle damage and oxidative stress, mediated by glutathione (GSH) in high-intensity aerobic exercise by increasing the 70-kDa heat shock proteins (HSP70) and heat shock factor 1 (HSF1). MAIN METHODS: Adult male Wistar rats were 8-week trained (60-min/day, 5 days/week) on a treadmill. During the last 21 days, the animals were supplemented with either l-alanyl-l-glutamine dipeptide (1.5 g/kg, DIP) or a solution containing the amino acids l-glutamine (1g/kg) and l-alanine (0.67 g/kg) in their free form (GLN+ALA) or water (controls). KEY FINDINGS: Plasma from both DIP- and GLN+ALA-treated animals showed higher l-glutamine concentrations and reduced ammonium, malondialdehyde, myoglobin and creatine kinase activity. In the soleus and gastrocnemius muscle of both supplemented groups, l-glutamine and GSH contents were increased and GSH disulfide (GSSG) to GSH ratio was attenuated (p<0.001). In the soleus muscle, cytosolic and nuclear HSP70 and HSF1 were increased by DIP supplementation. GLN+ALA group exhibited higher HSP70 (only in the nucleus) and HSF1 (cytosol and nucleus). In the gastrocnemius muscle, both supplementations were able to increase cytosolic HSP70 and cytosolic and nuclear HSF1. SIGNIFICANCE: In trained rats, oral supplementation with DIP or GLN+ALA solution increased the expression of muscle HSP70, favored muscle l-glutamine/GSH status and improved redox defenses, which attenuate markers of muscle damage, thus improving the beneficial effects of high-intensity exercise training.

Suplementações nutricionais e estresse oxidativo: implicações na atividade física e no esporte / Nutritional supplementation and oxidative stress: implications for physical activity and sport / Suplementación nutricional y estrés oxidativo: implicaciones para la actividad física y el deporte

Exercícios físicos associados a uma dieta balanceada são importantes fatores para a promoção da saúde. Contudo, a realização de exercícios físicos intensos e prolongados ou de caráter exaustivo pode promover inflamação crônica, overtraining e maior susceptibilidade a infecções. Sendo causa ou consequência, um dos fatores que contribuem para estes efeitos é o aumento exacerbado da síntese de compostos pró-oxidantes, conhecidos como espécies reativas do oxigênio (ERO) e nitrogênio (ERN). O aumento de ERO e ERN pode reduzir a capacidade antioxidante corporal, situação conhecida como estresse oxidativo. O estresse oxidativo tem sido relacionado como promotor de lesões a diversos constituintes celulares, principalmente sobre as membranas, efeito denominado como peroxidação lipídica. Para neutralizar os efeitos das ERO e ERN, o organismo dispõe do sistema de defesa antioxidante, localizado em diferentes compartimentos celulares e com funções diversas. Estudos têm, cada vez mais, demonstrado que o sistema antioxidante pode ser influenciado por intervenções nutricionais específicas, dentre as quais se incluem vitaminas, minerais, flavonóides e aminoácidos. Considerando o fato de muitas pessoas iniciarem a prática de exercícios físicos a cada dia, e que muitas destas ultrapassam seus limites, esta revisão visa abordar os principais sítios de síntese de ERO e ERN durante exercícios físicos, bem como possíveis estratégias nutricionais e seus mecanismos de ação sobre o sistema de defesa antioxidante.

Physical exercises associated with a balanced diet are important factors for health promotion. However intense and prolonged or strenuous exercise may promote chronic inflammation, overtraining and increased susceptibility to infections. Being cause or consequence, one of the factors that contribute to deleterious effects is exacerbated increase in the synthesis of pro-oxidant compounds, known as reactive oxygen species (ROS) and nitrogen species (RNS). The increase of ROS and RNS may reduce the body antioxidant capability, a condition known as oxidative stress. Oxidative stress has been implicated as a promoter of injuries to various cellular constituents, especially on the membranes, an effect known as lipid peroxidation. To attenuate the effects of ROS and RNS, the body has the antioxidant defense system, located in different cellular compartments and with different functions. Studies have increasingly shown that the antioxidant system can be influenced by specific nutritional interventions, among which are included vitamins, minerals, flavonoids and amino acids. Considering the fact that thousands of people engage in the practice of physical exercise every day, and that many of them go beyond their limits, this review aims to address the major sites of synthesis of ROS during exercise and nutrition strategies and their possible mechanisms action on the antioxidant defense system.

Ejercicios físicos asociados con una dieta equilibrada son factores importantes para la promoción de la salud. Sin embargo el ejercicio intenso y prolongado puede promover la inflamación crónica, el sobreentrenamiento y el aumento de la susceptibilidad a las infecciones. Siendo causa o consecuencia, uno de los factores que contribuyen a los efectos nocivos es el aumento exagerado de la síntesis de compuestos pro-oxidantes, conocidos como especies reactivas del oxígeno (ERO) y nitrógeno (ERN). El aumento de ERO y ERN puede reducir la capacidad antioxidante del cuerpo, una condición conocida como estrés oxidativo. El estrés oxidativo ha sido implicado como un promotor de las lesiones de varios componentes celulares, especialmente en las membranas, un efecto conocido como la peroxidación lipídica. Para atenuar los efectos de los ERO y ERN, el cuerpo tiene el sistema de defensa antioxidante, ubicado en diferentes compartimentos celulares y con diferentes funciones. Los estudios han demostrado cada vez que el sistema antioxidante puede ser influenciado por intervenciones nutricionales específicas, entre las que se incluyen vitaminas, minerales, flavonoides y aminoácidos. Teniendo en cuenta el hecho de que miles de personas participan en la práctica de ejercicio físico todos los días, y que muchos de estos van más allá de sus límites, esta revisión tiene como objetivo abordar los principales sitios de síntesis de ERO durante el ejercicio y estrategias de nutrición y sus mecanismos de acción en el sistema de defensa antioxidante.

Exercício físico e estresse oxidativo: mecanismos e efeitos / Exercise and oxidative stress: mechanisms and effects

Exercícios físicos associados a uma dieta balanceada são importantes fatores para a promoção da saúde. Contudo, a realização de exercícios físicos intensos e prolongados ou de caráter exaustivo podem promover inflamação crônica, overtraining e maior susceptibilidade de infecções. Sendo causa ou consequência, um dos fatores que contribuem para estes efeitos é o aumento exacerbado da síntese de compostos pró-oxidantes, conhecidos como espécies reativas do oxigênio (ERO). O aumento das ERO pode reduzir a capacidade antioxidante corporal, situação conhecida como estresse oxidativo. O estresse oxidativo tem sido relacionado ao aumento de lesões a diversos constituintes celulares, principalmente sobre as membranas, haja vista desencadear um processo de degeneração dos fosfolipídios, conhecido como peroxidação lipídica (PL). Dentre as fontes de síntese de ERO, induzidas pelo exercício físico estão às mitocôndrias, o processo de isquemia e reperfusão tecidual, a inflamação e a exacerbada liberação de íons metais de transição. Quando ocorrido cronicamente, o estresse oxidativo pode reduzir a massa e a força muscular, bem como, aumentar a gravidade de lesões às células, resultando em menor capacidade de recuperação. Deste modo, face essencial o conhecimento dos mecanismos e efeitos das ERO induzidas por exercícios físicos, bem como seus efeitos sobre o sistema antioxidante corporal.

Physical exercises associated with a balanced diet are important factors for health promotion. However intense and prolonged or strenuous exercise may promote chronic inflammation, overtraining and increased susceptibility to infections. Being cause or consequence, one of the factors that contribute to deleterious effects is exacerbated increase in the synthesis of pro-oxidant compounds, known as reactive oxygen species (ROS). The increase of ROS may reduce the body antioxidant capability, a condition known as oxidative stress. Oxidative stress has been implicated as a promoter of injuries to various cellular constituents, especially, on the membranes, due to trigger the degeneration of phospholipids, an effect known as lipid peroxidation. Among the sources for the synthesis of ROS induced by exercise is the mitochondria, the process of tissue ischemia and reperfusion, inflammation and exaggerated release of transition metal ions. When occurred chronically, oxidative stress can reduce muscle mass and strength, as well as increase the severity of injuries to the cells, resulting in lower resilience. Thus, given the essential knowledge of the mechanisms and effects of ROS induced by exercise, as well as its effects on the antioxidant system of body.

Glutamina: aspectos bioquímicos, metabólicos, moleculares e suplementação / Glutamine: biochemical, metabolic, molecular aspects and supplementation

A glutamina é o aminoácido livre mais abundante no plasma e no tecido muscular. Nutricionalmente é classificada como um aminoácido não essencial, uma vez que pode ser sintetizada pelo organismo a partir de outros aminoácidos. A glutamina está envolvida em diferentes funções, tais como a proliferação e desenvolvimento de células, o balanço acidobásico, o transporte da amônia entre os tecidos, a doação de esqueletos de carbono para a gliconeogênese, a participação no sistema antioxidante e outras. Por meio de técnicas de biologia molecular, estudos demonstram que a glutamina pode também influenciar diversas vias de sinalização celular, em especial a expressão de proteínas de choque térmico (HSPs). As HSPs contribuem para a manutenção da homeostasia da célula na presença de agentes estressores, tais como as espécies reativas de oxigênio (ERO). Em situações de elevado catabolismo muscular, como após exercícios físicos intensos e prolongados, a concentração de glutamina pode tornar-se reduzida. A menor disponibilidade desse aminoácido pode diminuir a resistência da célula a lesões, levando a processos de apoptose celular. Por essas razões, a suplementação com L-glutamina, tanto na forma livre, quanto como dipeptídeo, tem sido investigada. Alguns aspectos bioquímicos, metabólicos e mecanismos moleculares da glutamina, bem como os efeitos de sua suplementação, são abordados no presente trabalho.

Glutamine is the most frequent free amino acid in the serum and muscular tissue. Nutritionally, it is classified as a non-essential amino acid, once it can be synthesized by the body from other amino acids. Glutamine is involved in different functions, such as cell proliferation and development, basic acid balance, ammonia transportation between tissues, carbon skeleton donation to the gluconeogenesis, participation in the antioxidant system, among others. Molecular biology techniques show that it may also influence several cell signaling ways, especially the expression of heat shock proteins (HSP). The HSPs contribute to the maintenance of the cellular homeostasis in the presence of stress agents such as oxygen reactive species (ORE). In situations of high cellular catabolism, as after intense and prolonged physical exercises, the glutamine concentration may become reduced. Lower availability of this amino acid may decrease the cell resistance to injuries, leading to cellular apoptosis processes. Therefore, L-glutamine supplementation either in free form or as dipeptide has been investigated. Some biochemical and metabolic aspects, molecular mechanism of glutamine, as well as the effects of its supplementation are approached in the present article.