Changes in Skeletal Muscle Protein Metabolism Signaling Induced by Glutamine Supplementation and Exercise.

AIM: To evaluate the effects of resistance exercise training (RET) and/or glutamine supplementation (GS) on signaling protein synthesis in adult rat skeletal muscles. METHODS: The following groups were studied: (1) control, no exercise (C); (2) exercise, hypertrophy resistance exercise training protocol (T); (3) no exercise, supplemented with glutamine (G); and (4) exercise and supplemented with glutamine (GT). The rats performed hypertrophic training, climbing a vertical ladder with a height of 1.1 m at an 80° incline relative to the horizontal with extra weights tied to their tails. The RET was performed three days a week for five weeks. Each training session consisted of six ladder climbs. The extra weight load was progressively increased for each animal during each training session. The G groups received daily L-glutamine by gavage (one g per kilogram of body weight per day) for five weeks. The C group received the same volume of water during the same period. The rats were euthanized, and the extensor digitorum longus (EDL) muscles from both hind limbs were removed and immediately weighed. Glutamine and glutamate concentrations were measured, and histological, signaling protein contents, and mRNA expression analyses were performed. RESULTS: Supplementation with free L-glutamine increased the glutamine concentration in the EDL muscle in the C group. The glutamate concentration was augmented in the EDL muscles from T rats. The EDL muscle mass did not change, but a significant rise was reported in the cross-sectional area (CSA) of the fibers in the three experimental groups. The levels of the phosphorylated proteins (pAkt/Akt, pp70S6K/p70S6K, p4E-BP1/4E-BP1, and pS6/S6 ratios) were significantly increased in EDL muscles of G rats, and the activation of p4E-BP1 was present in T rats. The fiber CSAs of the EDL muscles in T, G, and GT rats were increased compared to the C group. These changes were accompanied by a reduction in the 26 proteasome activity of EDL muscles from T rats. CONCLUSION: Five weeks of GS and/or RET induced muscle hypertrophy, as indicated by the increased CSAs of the EDL muscle fibers. The increase in CSA was mediated via the upregulated phosphorylation of Akt, 4E-BP1, p70S6k, and S6 in G animals and 4E-BP1 in T animals. In the EDL muscles from T animals, a decrease in proteasome activity, favoring a further increase in the CSA of the muscle fibers, was reported.

Changes in skeletal muscle protein metabolism signaling induced by glutamine supplementation and esxercise

Aim: To evaluate the effects of resistance exercise training (RET) and/or glutamine supplementation (GS) on signaling protein synthesis in adult rat skeletal muscles. Methods: The following groups were studied: (1) control, no exercise (C); (2) exercise, hypertrophy resistance exercise training protocol (T); (3) no exercise, supplemented with glutamine (G); and (4) exercise and supplemented with glutamine (GT). The rats performed hypertrophic training, climbing a vertical ladder with a height of 1.1 m at an 80° incline relative to the horizontal with extra weights tied to their tails. The RET was performed three days a week for five weeks. Each training session consisted of six ladder climbs. The extra weight load was progressively increased for each animal during each training session. The G groups received daily L-glutamine by gavage (one g per kilogram of body weight per day) for five weeks. The C group received the same volume of water during the same period. The rats were euthanized, and the extensor digitorum longus (EDL) muscles from both hind limbs were removed and immediately weighed. Glutamine and glutamate concentrations were measured, and histological, signaling protein contents, and mRNA expression analyses were performed. Results: Supplementation with free L-glutamine increased the glutamine concentration in the EDL muscle in the C group. The glutamate concentration was augmented in the EDL muscles from T rats. The EDL muscle mass did not change, but a significant rise was reported in the cross-sectional area (CSA) of the fibers in the three experimental groups. The levels of the phosphorylated proteins (pAkt/Akt, pp70S6K/p70S6K, p4E-BP1/4E-BP1, and pS6/S6 ratios) were significantly increased in EDL muscles of G rats, and the activation of p4E-BP1 was present in T rats. The fiber CSAs of the EDL muscles in T, G, and GT rats were increased compared to the C group. These changes were accompanied by a reduction in the 26 proteasome activity of EDL muscles from T rats. Conclusion: Five weeks of GS and/or RET induced muscle hypertrophy, as indicated by the increased CSAs of the EDL muscle fibers. The increase in CSA was mediated via the upregulated phosphorylation of Akt, 4E-BP1, p70S6k, and S6 in G animals and 4E-BP1 in T animals. In the EDL muscles from T animals, a decrease in proteasome activity, favoring a further increase in the CSA of the muscle fibers, was reported.

Endoplasmic Reticulum Stress and Autophagy Markers in Soleus Muscle Disuse-Induced Atrophy of Rats Treated with Fish Oil.

Endoplasmic reticulum stress (ERS) and autophagy pathways are implicated in disuse muscle atrophy. The effects of high eicosapentaenoic (EPA) or high docosahexaenoic (DHA) fish oils on soleus muscle ERS and autophagy markers were investigated in a rat hindlimb suspension (HS) atrophy model. Adult Wistar male rats received daily by gavage supplementation (0.3 mL per 100 g b.w.) of mineral oil or high EPA or high DHA fish oils (FOs) for two weeks. Afterward, the rats were subjected to HS and the respective treatments concomitantly for an additional two-week period. After four weeks, we evaluated ERS and autophagy markers in the soleus muscle. Results were analyzed using two-way analysis of variance (ANOVA) and Bonferroni post hoc test. Gastrocnemius muscle ω-6/ω-3 fatty acids (FAs) ratio was decreased by both FOs indicating the tissue incorporation of omega-3 fatty acids. HS altered (p < 0.05) the protein content (decreasing total p38 and BiP and increasing p-JNK2/total JNK2 ratio, and caspase 3) and gene expressions (decreasing BiP and increasing IRE1 and PERK) of ERS and autophagy (decreasing Beclin and increasing LC3 and ATG14) markers in soleus. Both FOs attenuated (p < 0.05) the increase in PERK and ATG14 expressions induced by HS. Thus, both FOs could potentially attenuate ERS and autophagy in skeletal muscles undergoing atrophy.

Endoplasmic reticulum stress and autophagy markers in Soleus muscle disuse-induced atrophy of rats treated with fish oil

Endoplasmic reticulum stress (ERS) and autophagy pathways are implicated in disuse muscle atrophy. The effects of high eicosapentaenoic (EPA) or high docosahexaenoic (DHA) fish oils on soleus muscle ERS and autophagy markers were investigated in a rat hindlimb suspension (HS) atrophy model. Adult Wistar male rats received daily by gavage supplementation (0.3 mL per 100 g b.w.) of mineral oil or high EPA or high DHA fish oils (FOs) for two weeks. Afterward, the rats were subjected to HS and the respective treatments concomitantly for an additional two-week period. After four weeks, we evaluated ERS and autophagy markers in the soleus muscle. Results were analyzed using two-way analysis of variance (ANOVA) and Bonferroni post hoc test. Gastrocnemius muscle ω-6/ω-3 fatty acids (FAs) ratio was decreased by both FOs indicating the tissue incorporation of omega-3 fatty acids. HS altered (p < 0.05) the protein content (decreasing total p38 and BiP and increasing p-JNK2/total JNK2 ratio, and caspase 3) and gene expressions (decreasing BiP and increasing IRE1 and PERK) of ERS and autophagy (decreasing Beclin and increasing LC3 and ATG14) markers in soleus. Both FOs attenuated (p < 0.05) the increase in PERK and ATG14 expressions induced by HS. Thus, both FOs could potentially attenuate ERS and autophagy in skeletal muscles undergoing atrophy.

Addition of Caffeine to a Carbohydrate Feeding Strategy Prior to Intermittent Exercise.

The ergogenic effect of caffeine is well established, although no investigations providing a high carbohydrate feeding strategy (pre-exercise meal=2 g/kg BM) co-ingested with caffeine exist for soccer. This investigation examines the effect of caffeine in addition to a pre-exercise carbohydrate meal and drink mid-way through a soccer simulation. Eight recreational soccer players completed an 85-minute soccer simulation followed by an exercise capacity test (Yo-yo Intermittent Endurance test level 2) on two occasions. Prior to exercise participants consumed a high carbohydrate meal, with placebo or 5 mg/kg BM-1 caffeine. No significant performance effect was identified (p=0.099) despite a 12.8% (109 m) improvement in exercise capacity following caffeine. Rates of carbohydrate and fat oxidation did not differ between conditions and nor were differences apparent for plasma glucose, fatty acids, glycerol, ß-hydroxybutyrate (p>0.05). However, an increase in lactate was observed for caffeine (p=0.039). A significant condition effect on rating of perceived exertion was identified (p<0.001), with the overall mean for the protocol lowered to 11.7±0.9 au for caffeine compared to 12.8±1.3 au. Caffeine supplementation with a carbohydrate feeding strategy failed to affect metabolic and metabolite responses, although reductions in perception of exercise were observed. While a 12.8% increase in exercise capacity was noted the findings were not significant, possibly due to the small sample size.

Glutamine supplementation versus functional overload in extensor digitorum longus muscle hypertrophy

Background Glutamine levels directly associate with total protein content in cultured skeletal muscle cells, whereas glutamine supplementation enhances skeletal muscle mass in catabolic experimental conditions. Methods We compared the effect of glutamine administration on Extensor Digitorum Longus muscle (EDL) weight, fiber cross-sectional area (CSA), contractile activity, and protein metabolism signaling with a functional overload-induced skeletal muscle hypertrophy protocol. Results Glutamine supplementation raised the predominance of EDL muscle fibers with CSA between 1001 and 2000 μm2 (49.7 %), the p-4E-BP1/total 4E-BP1 ratio, and the effect of overload on resistance to fatigue. The proportion of the EDL muscle fiber CSA distribution for the combination of both treatments was similar to that induced by overload or glutamine separately; 54.3 % muscle fibers with CSA between 1001 and 2000 μm². Glutamine supplementation did not markedly affect the changes induced by overload on protein synthesis signaling pathways, except for a further increase of the p-4E-BP1/total 4E-BP1 ratio. Conclusions The effect of glutamine on EDL muscle fiber CSA distribution and protein synthesis signaling mimicked the response to overload. The association of glutamine and overload induced EDL muscle hypertrophy further increased the resistance to fatigue.

Effectiveness of short-term physical agents treatment on macroscopic morphology in patients with plaque psoriasis / Efectividad del tratamiento a corto plazo de los agentes físicos en la morfología macroscópica de las placas de psoriasis

Psoriasis is a chronic inflammatory disease that presents skin rashes which can arise through plaques. The aim of this work was to compare the effectiveness of short-term physical agents treatment on macroscopic morphology (area and erythema) in patients with plaque psoriasis. This prospective randomized experimental study included fourteen subjects, medically diagnosed with psoriasis, with more than one plaque in the skin and voluntarily without topical treatment. All subjects completed the study that consisted of 12 treatment sessions divided in control (C), artificial balneotherapy (AB), phototherapy (PT) or balneophototherapy (BPT) groups. After session 12, there was a significant reduction of the plaque area by all treatments when compared to C group and BPT was the most effective one. However, only AB and PT presented a reduction of erythema. Regarding severity, 9 patients changed to a lower category on the PASI test, and 5 of them maintained a mild psoriasis, but lowered their score. Finally, 13 of 14 subjects improved their quality of life. The physical agents used reduced the severity of psoriasis and improved quality of life of patients after 12 sessions of treatment during a onemonth period. The BPT was the more effective in controlling psoriasis by diminishing its area and PT by attenuating the erythema.

La Psoriasis es una enfermedad inflamatoria crónica que presenta irritación cutánea que puede derivar a placas. El objetivo de este trabajo fue comparar la efectividad del tratamiento a corto plazo con agentes físicos en la morfología macroscópica (área y eritema) en pacientes con placas de psoriasis. Estudio experimental, prospectivo, randomizado. Catorce sujetos participaron con diagnóstico médico de psoriasis, con más de una placa en la piel y sin tener tratamiento tópico de forma voluntaria. Todos los sujetos completaron el estudio, el cual consistió de 12 sesiones de tratamiento dividido en grupo control (C), BA, FT y BFA. Posterior a la sesión 12, se observó una reducción significativa en toda el área de las placas que recibieron tratamiento al compararlas al grupo C y el grupo BFA fue el más efectivo. Sin embargo, solo los grupos BA y FT presentaron una reducción del eritema. Respecto a la severidad, 9 pacientes cambiaron de la baja categoría en el test de PASI y 5 de ellos se mantuvieron en el nivel medio, pero disminuyeron su puntaje. Finalmente, 13 de 14 sujetos mejoraron su calidad de vida. Los agentes físicos usados redujeron la severidad de la psoriasis y mejoraron la calidad de vida de los pacientes después de 12 sesiones de tratamiento durante el período de un mes. La BFA fue la más efectiva en controlar la psoriasis por la disminución en el área y la FT por la atenuación del eritema.

Oral L-glutamine pretreatment attenuates skeletal muscle atrophy induced by 24-h fasting in mice.

L-Glutamine (L-Gln) supplementation has been pointed out as an anticatabolic intervention, but its effects on protein synthesis and degradation signaling in skeketal muscle are still poorly known. The effects of L-Gln pretreatment (1 g kg-1 day-1 body weight for 10 days) on muscle fiber cross-sectional area (CSA), amino acid composition (measured by LC-MS/MS) and protein synthesis (Akt-mTOR) and degradation (ubiquitin ligases) signaling in soleus and extensor digitorum longus (EDL) muscles in 24-h-fasted mice were investigated. The fiber CSA of EDL muscle was not different between the L-Gln-fasted and L-Gln-fed groups. This finding was associated with reduced contents of L-Leu and L-Iso and activation of protein synthesis signaling (p-RPS6Ser240/244 and Akt-mTOR). The spectrum of soleus muscle fiber CSA distribution was larger in L-Gln-fasted as compared with placebo-fasted mice. This effect of L-Gln pretreatment was associated with changes in red fibers L-Gln metabolism as indicated by increased intracellular L-glutamine/L-glutamate ratio, L-aspartate and GABA levels. L-Gln supplementation reduced fasting-induced mass loss in tibialis anterior and gastrocnemius muscles. Evidence is presented that pretreatment with L-glutamine attenuates skeletal muscle atrophy induced by 24-h fasting through mechanisms that vary with the muscle fiber type.

In Vivo Electrical Stimulation for the Assessment of Skeletal Muscle Contractile Function in Murine Models.

Skeletal muscle electrical stimulation is commonly used for clinical purposes, assisting recovery, preservation, or even improvement of muscle mass and function in healthy and pathological conditions. Additionally, it is a useful research tool for evaluation of skeletal muscle contractile function. It may be applied in vitro, using cell culture or isolated fibers/muscles, and in vivo, using human subjects or animal models (neuromuscular electrical stimulation - NMES). This chapter focuses on the electrical stimulation of the sciatic nerve as a research method for evaluation of the contractile properties of murine hind limb muscles. Variations of this protocol allow for the assessment of muscle force, fatigue resistance, contraction and relaxation times, and can be used as a model of contraction-induced muscle injury, reactive oxygen species production, and muscle adaptation to contractile activity.

Hypertrophy Stimulation at the Onset of Type I Diabetes Maintains the Soleus but Not the EDL Muscle Mass in Wistar Rats.

Diabetes mellitus induces a reduction in skeletal muscle mass and strength. Strength training is prescribed as part of treatment since it improves glycemic control and promotes increase of skeletal muscle mass. The mechanisms involved in overload-induced muscle hypertrophy elicited at the establishment of the type I diabetic state was investigated in Wistar rats. The purpose was to examine whether the overload-induced hypertrophy can counteract the hypotrophy associated to the diabetic state. The experiments were performed in oxidative (soleus) or glycolytic (EDL) muscles. PI3K/Akt/mTOR protein synthesis pathway was evaluated 7 days after overload-induced hypertrophy of soleus and of EDL muscles. The mRNA expression of genes associated with different signaling pathways that control muscle hypertrophy was also evaluated: mechanotransduction (FAK), Wnt/ß-catenin, myostatin, and follistatin. The soleus and EDL muscles when submitted to overload had similar hypertrophic responses in control and diabetic animals. The increase of absolute and specific twitch and tetanic forces had the same magnitude as muscle hypertrophic response. Hypertrophy of the EDL muscle from diabetic animals mostly involved mechanical loading-stimulated PI3K/Akt/mTOR pathway besides the reduced activation of AMP-activated protein kinase (AMPK) and decrease of myostatin expression. Hypertrophy was more pronounced in the soleus muscle of diabetic animals due to a more potent activation of rpS6 and increased mRNA expression of insulin-like growth factor-1 (IGF-1), mechano-growth factor (MGF) and follistatin, and decrease of myostatin, MuRF-1 and atrogin-1 contents. The signaling changes enabled the soleus muscle mass and force of the diabetic rats to reach the values of the control group.

Effects of high EPA and high DHA fish oils on changes in signaling associated with protein metabolism induced by hindlimb suspension in rats.

The effects of either eicosapentaenoic (EPA)- or docosahexaenoic (DHA)-rich fish oils on hindlimb suspension (HS)-induced muscle disuse atrophy were compared. Daily oral supplementations (0.3 mL/100 g b.w.) with mineral oil (MO) or high EPA or high DHA fish oils were performed in adult rats. After 2 weeks, the animals were subjected to HS for further 2 weeks. The treatments were maintained alongside HS At the end of 4 weeks, we evaluated: body weight gain, muscle mass and fat depots, composition of fatty acids, cross-sectional areas (CSA) of the soleus muscle and soleus muscle fibers, activities of cathepsin L and 26S proteasome, and content of carbonylated proteins in the soleus muscle. Signaling pathway activities associated with protein synthesis (Akt, p70S6K, S6, 4EBP1, and GSK3-beta) and protein degradation (atrogin-1/MAFbx, and MuRF1) were evaluated. HS decreased muscle mass, CSA of soleus muscle and soleus muscle fibers, and altered signaling associated with protein synthesis (decreased) and protein degradation (increased). The treatment with either fish oil decreased the ratio of omega-6/omega-3 fatty acids and changed protein synthesis-associated signaling. EPA-rich fish oil attenuated the changes induced by HS on 26S proteasome activity, CSA of soleus muscle fibers, and levels of p-Akt, total p70S6K, p-p70S6K/total p70S6K, p-4EBP1, p-GSK3-beta, p-ERK2, and total ERK 1/2 proteins. DHA-rich fish oil attenuated the changes induced by HS on p-4EBP1 and total ERK1 levels. The effects of EPA-rich fish oil on protein synthesis signaling were more pronounced. Both EPA- and DHA-rich fish oils did not impact skeletal muscle mass loss induced by non-inflammatory HS.

Contractile function recovery in severely injured gastrocnemius muscle of rats treated with either oleic or linoleic acid.

NEW FINDINGS: What is the central question of this study? Oleic and linoleic acids modulate fibroblast proliferation and myogenic differentiation in vitro. However, their in vivo effects on muscle regeneration have not yet been examined. We investigated the effects of either oleic or linoleic acid on a well-established model of muscle regeneration after severe laceration. What is the main finding and its importance? We found that linoleic acid increases fibrous tissue deposition and impairs muscle regeneration and recovery of contractile function, whereas oleic acid has the opposite effects in severely injured gastrocnemius muscle, suggesting that linoleic acid has a harmful effect and oleic acid a potential therapeutic effect on muscle regeneration. Oleic and linoleic acids control fibroblast proliferation and myogenic differentiation in vitro; however, there was no study in skeletal muscle in vivo. The aim of this study was to evaluate the effects of either oleic or linoleic acid on the fibrous tissue content (collagen deposition) of muscle and recovery of contractile function in rat gastrocnemius muscle after being severely injured by laceration. Rats were supplemented with either oleic or linoleic acid for 4 weeks after laceration [0.44 g (kg body weight)-1 day-1 ]. Muscle injury led to an increase in oleic-to-stearic acid and palmitoleic-to-palmitic acid ratios, suggesting an increase in Δ9 desaturase activity. Increased fibrous tissue deposition and reduced isotonic and tetanic specific forces and resistance to fatigue were observed in the injured muscle. Supplementation with linoleic acid increased the content of eicosadienoic (20:2, n-6) and arachidonic (20:4, n-6) acids, reduced muscle mass and fibre cross-sectional areas, increased fibrous tissue deposition and further reduced the isotonic and tetanic specific forces and resistance to fatigue induced by laceration. Supplementation with oleic acid increased the content of docosahexaenoic acid (22:6, n-3) and abolished the increase in fibrous tissue area and the decrease in isotonic and tetanic specific forces and resistance to fatigue induced by muscle injury. We concluded that supplementation with linoleic acid impairs muscle regeneration and increases fibrous tissue deposition, resulting in impaired recovery of contractile function. Oleic acid supplementation reduced fibrous tissue deposition and improved recovery of contractile function, attenuating the tissue damage caused by muscle injury.

Heat transfer by three types of hot pack and its implication on the flexibility of the lower back: a randomized, controlled trial / Transferência de calor por três tipos de compressas quentes e sua implicação na flexibilidade da região lombar: ensaio clínico randomizado e controlado / Transferencia de calor por tres tipos de compresas calientes y su implicación en la flexibilidad en la región lumbar: ensayo clínico aleatorio y controlado

ABSTRACT The use of hot pack is a common superficial thermotherapy strategy and one of its benefits is the increase of muscle flexibility. However, there is a lack of information about the effects of the heat pack alone, without being used in association with other therapeutic interventions, in the flexibility of the lumbar region. The aim of this study was to compare the effects generated by the application of three different pack on the flexibility of the lower backs of healthy students. Three sessions of 15 minutes of superficial heat through a hot pack (moist heat pack-MHP, seed pack-SP or gel pack-GP) were applied to the lower back. Pack and lower back temperatures and erythema were registered every 5 minutes. A Schober test was performed before the first session and after the third session. After 15 minutes of treatment, pack temperature was higher in the SP group. At the same time, lumbar temperature was lower in the GP group. The heat treatment also increased erythema in the lower back for all three groups. There was a significant increase in intragroup flexibility as assessed by the Schober Test for all groups. There are significant differences in the effect generated between the three types of pack on the flexibility of the lower back. The MHP was able to transfer more heat to the lumbar area and provided a more pronounced increase in the flexibility of lower back tissues.

RESUMO O uso de compressas quentes é uma estratégia de termoterapia superficial amplamente utilizada e um de seus benefícios é o aumento da flexibilidade muscular. Porém, existem poucas informações sobre os efeitos das compressas quentes, quando não associadas a outras intervenções terapêuticas, na flexibilidade da região lombar. O objetivo do seguinte trabalho foi comparar os efeitos gerados pela aplicação de três tipos diferentes de compressas quentes na flexibilidade da região lombar de estudantes saudáveis. Três sessões de 15 minutos de calor superficial aplicado através de compressas quentes (compressa úmida quente, compressa de sementes e compressa de gel). A temperatura da compressa e da região lombar e a ocorrência de eritema foram registradas a cada 5 minutos. O teste de Schober foi realizado antes da primeira e após a última sessão. Após 15 minutos de tratamento, a compressa de sementes apresentou maior temperatura final. No mesmo período, a menor temperatura lombar foi obtida pela compressa de gel. O tratamento com os três tipos de compressa aumentou a ocorrência de eritema e causou aumento significativo da flexibilidade da região lombar avaliada pelo teste de Schober. Existem diferenças significativas no efeito gerado pelos três tipos de compressas quentes sobre a flexibilidade da região lombar. A compressa úmida quente proporcionou maior transferência de calor para a região lombar e propiciou um aumento mais pronunciado da flexibilidade da região lombar.

RESUMEN El empleo de compresas calientes es muy utilizado por la termoterapia superficial como forma de aumentar la flexibilidad muscular. Pero son pocas las informaciones sobre sus efectos, cuando asociadas a otras intervenciones terapéuticas, en la flexibilidad de la región lumbar. Este estudio tiene el objeto de comparar los efectos de la aplicación de tres tipos distintos de compresas calientas en la flexibilidad de la región lumbar en estudiantes saludables. Fueron tres sesiones de quince minutos de calor superficial aplicadas a través de compresas calientes (compresa húmeda, compresa de semillas y compresa de gel) en la región lumbar de estos participantes. Se registraban cada cinco minutos la temperatura de la compresa y de la región lumbar y la existencia de eritema. Se empleó la prueba de Schober realizada antes de la primera y después de la última sesión. Tras quince minutos de tratamiento, la compresa de semillas presentó una temperatura final mayor. En este mismo periodo, la menor temperatura lumbar la registró la compresa de gel. El tratamiento con tres tipos de compresas aumentó la existencia de eritema y el significativo aumento de la flexibilidad de la región lumbar, evaluado por la prueba de Schober. Diferencias significativas ocurrieron con el empleo de los tres tipos de compresas calientes sobre la flexibilidad de la región lumbar. La compresa húmeda caliente tuvo una transferencia de calor para la región lumbar mayor, por lo que aumentó más la flexibilidad de la región evaluada.

Loss of the anorexic response to systemic 5-aminoimidazole-4-carboxamide-1-ß-D-ribofuranoside administration despite reducing hypothalamic AMP-activated protein kinase phosphorylation in insulin-deficient rats.

This study tested whether chronic systemic administration of 5-aminoimidazole-4-carboxamide-1-ß-D-ribofuranoside (AICAR) could attenuate hyperphagia, reduce lean and fat mass losses, and improve whole-body energy homeostasis in insulin-deficient rats. Male Wistar rats were first rendered diabetic through streptozotocin (STZ) administration and then intraperitoneally injected with AICAR for 7 consecutive days. Food and water intake, ambulatory activity, and energy expenditure were assessed at the end of the AICAR-treatment period. Blood was collected for circulating leptin measurement and the hypothalami were extracted for the determination of suppressor of cytokine signaling 3 (SOCS3) content, as well as the content and phosphorylation of AMP-kinase (AMPK), acetyl-CoA carboxylase (ACC), and the signal transducer and activator of transcription 3 (STAT3). Rats were thoroughly dissected for adiposity and lean body mass (LBM) determinations. In non-diabetic rats, despite reducing adiposity, AICAR increased (∼1.7-fold) circulating leptin and reduced hypothalamic SOCS3 content and food intake by 67% and 25%, respectively. The anorexic effect of AICAR was lost in diabetic rats, even though hypothalamic AMPK and ACC phosphorylation markedly decreased in these animals. Importantly, hypothalamic SOCS3 and STAT3 levels remained elevated and reduced, respectively, after treatment of insulin-deficient rats with AICAR. Diabetic rats were lethargic and displayed marked losses of fat and LBM. AICAR treatment increased ambulatory activity and whole-body energy expenditure while also attenuating diabetes-induced fat and LBM losses. In conclusion, AICAR did not reverse hyperphagia, but it promoted anti-catabolic effects on skeletal muscle and fat, enhanced spontaneous physical activity, and improved the ability of rats to cope with the diabetes-induced dysfunctional alterations in glucose metabolism and whole-body energy homeostasis.

Short-term creatine supplementation decreases reactive oxygen species content with no changes in expression and activity of antioxidant enzymes in skeletal muscle.

The effect of short-term creatine (Cr) supplementation upon content of skeletal muscle-derived-reactive oxygen species (ROS) was investigated. Wistar rats were supplemented with Cr (5 g/kg BW) or vehicle, by gavage, for 6 days. Soleus and extensor digitorum longus (EDL) muscles were removed and incubated for evaluation of ROS content using Amplex-UltraRed reagent. The analysis of expression and activity of antioxidant enzymes (superoxide dismutase 1 and 2, catalase and glutathione peroxidase) were performed. Direct scavenger action of Cr on superoxide radical and hydrogen peroxide was also investigated. Short-term Cr supplementation attenuated ROS content in both soleus and EDL muscles (by 41 and 33.7%, respectively). Cr supplementation did not change expression and activity of antioxidant enzymes. Basal TBARS content was not altered by Cr supplementation. In cell-free experiments, Cr showed a scavenger effect on superoxide radical in concentrations of 20 and 40 mM, but not on hydrogen peroxide. These results indicate that Cr supplementation decreases ROS content in skeletal muscle possibly due to a direct action of Cr molecule on superoxide radical.

Metabolic and functional effects of beta-hydroxy-beta-methylbutyrate (HMB) supplementation in skeletal muscle.

Beta-hydroxy-beta-methylbutyrate (HMB) is a metabolite derived from leucine. The anti-catabolic effect of HMB is well documented but its effect upon skeletal muscle strength and fatigue is still uncertain. In the present study, male Wistar rats were supplemented with HMB (320 mg/kg per day) for 4 weeks. Placebo group received saline solution only. Muscle strength (twitch and tetanic force) and resistance to acute muscle fatigue of the gastrocnemius muscle were evaluated by direct electrical stimulation of the sciatic nerve. The content of ATP and glycogen in red and white portions of gastrocnemius muscle were also evaluated. The effect of HMB on citrate synthase (CS) activity was also investigated. Muscle tetanic force was increased by HMB supplementation. No change was observed in time to peak of contraction and relaxation time. Resistance to acute muscle fatigue during intense contractile activity was also improved after HMB supplementation. Glycogen content was increased in both white (by fivefold) and red (by fourfold) portions of gastrocnemius muscle. HMB supplementation also increased the ATP content in red (by twofold) and white (1.2-fold) portions of gastrocnemius muscle. CS activity was increased by twofold in red portion of gastrocnemius muscle. These results support the proposition that HMB supplementation have marked change in oxidative metabolism improving muscle strength generation and performance during intense contractions.

Effect of short-term creatine supplementation on markers of skeletal muscle damage after strenuous contractile activity.

The protective effect of short-term creatine supplementation (CrS) upon markers of strenuous contractile activity-induced damage in human and rat skeletal muscles was investigated. Eight Ironman triathletes were randomized into the placebo (Pl; n = 4) and creatine-supplemented (CrS; n = 4) groups. Five days prior to the Ironman competition, the CrS group received creatine monohydrate (20 g day(-1)) plus maltodextrin (50 g) divided in two equal doses. The Pl group received maltodextrin (50 g day(-1)) only. The effect of CrS (5 g day(-1)/kg body weight for 5 days) was also evaluated in a protocol of strenuous contractile activity induced by electrical stimulation in rats. Blood samples were collected before and 36 and 60 h after the competition and were used to determine plasma activities of creatine kinase (CK), lactate dehydrogenase (LDH), aldolase (ALD), glutamic oxaloacetic acid transaminase (GOT), glutamic pyruvic acid transaminase (GPT), and C-reactive protein (CRP) level. In rats, plasma activities of CK and LDH, muscle vascular permeability (MVP) using Evans blue dye, muscle force and fatigue were evaluated. Activities of CK, ALD, LDH, GOT, GTP, and levels of CRP were increased in the Pl group after the competition as compared to basal values. CrS decreased plasma activities of CK, LDH, and ALD, and prevented the rise of GOT and GPT plasma activities. In rats, CrS delayed the fatigue, preserved the force, and prevented the rise of LDH and CK plasma activities and MVP in the gastrocnemius muscle. CrS presented a protective effect on muscle injury induced by strenuous contractile activities.