Humberto Nicastro

Potential antiproteolytic effects of L-leucine: observations of in vitro and in vivo studies

The purpose of present review is to describe the effect of leucine supplementation on skeletal muscle proteolysis suppression in both in vivo and in vitro studies. Most studies, using in vitro methodology, incubated skeletal muscles with leucine with different doses and the results suggests that there is a dose-dependent effect. The same responses can be observed in in vivo studies. Importantly, the leucine effects on skeletal muscle protein synthesis are not always connected to the inhibition of skeletal muscle proteolysis. As a matter of fact, high doses of leucine incubation can promote suppression of muscle proteolysis without additional effects on protein synthesis, and low leucine doses improve skeletal muscle protein ynthesis but have no effect on skeletal muscle proteolysis. These research findings may have an important clinical relevancy, because muscle loss in atrophic states would be reversed by specific leucine supplementation doses. Additionally, it has been clearly demonstrated that leucine administration suppresses skeletal muscle proteolysis in various catabolic states. Thus, if protein metabolism changes during different atrophic conditions, it is not surprising that the leucine dose-effect relationship must also change, according to atrophy or pathological state and catabolism magnitude. In conclusion, leucine has a potential role on attenuate skeletal muscle proteolysis. Future studies will help to sharpen the leucine efficacy on skeletal muscle protein degradation during several atrophic states.

An overview of the therapeutic effects of leucine supplementation on skeletal muscle under atrophic conditions

The characterization of the mechanisms underlying skeletal muscle atrophy under different conditions has been a constant focus of research. Among anti-atrophic therapies, amino acid supplementation, particularly with leucine, has received a lot of attention. Supplementation has been shown to have remarkable effects on muscle remodeling through protein turnover modulation. This may then impact physiological parameters related to muscle function, and even quality of life. In light of this, leucine supplementation could be a useful therapy for mitigating the atrophic effects of catabolic conditions. The purpose of this review is to present the major results of human studies evaluating the effects of leucine supplementation on structure and function of skeletal muscle in atrophic conditions such as muscle disuse, sarcopenia, and cancer.

Does Branched-Chain Amino Acids Supplementation Modulate Skeletal Muscle Remodeling through Inflammation Modulation? Possible Mechanisms of Action

Skeletal muscle protein turnover is modulated by intracellular signaling pathways involved in protein synthesis, degradation, and inflammation. The proinflammatory status of muscle cells, observed in pathological conditions such as cancer, aging, and sepsis, can directly modulate protein translation initiation and muscle proteolysis, contributing to negative protein turnover. In this context, branched-chain amino acids (BCAAs), especially leucine, have been described as a strong nutritional stimulus able to enhance protein translation initiation and attenuate proteolysis. Furthermore, under inflammatory conditions, BCAA can be transaminated to glutamate in order to increase glutamine synthesis, which is a substrate highly consumed by inflammatory cells such as macrophages. The present paper describes the role of inflammation on muscle remodeling and the possible metabolic and cellular effects of BCAA supplementation in the modulation of inflammatory status of skeletal muscle and the consequences on protein synthesis and degradation.

Glucocorticoids: Extensive physiological actions modulated through multiple mechanisms of gene regulation

Glucocorticoid hormones are important regulators of several physiological processes. Despite having been initially named based on their role in glucose metabolism, glucocorticoids are also fundamental in the regulation of developmental, metabolic, and neurobiological processes, as well as several other biological functions. Due to their involvement in a diverse array of biological pathways, its wide spectrum of action, it is predicted that a wide range of genes may have their expression regulated by the activated glucocorticoid receptor (GR). In fact, it has been demonstrated that in addition to the regulation of several effectors genes, the expression of the gene encoding for GR itself is regulated by physiological stimuli and fine‐tuning mechanisms. Importantly, such generalized effector responses and fine‐tuning responses seem to be largely mediated by mechanisms of gene regulation. Therefore, this review aims to describe the mechanisms of gene regulation by glucocorticoid hormones, which are capable of regulating differential gene transcription, within a physiological context. From this discussion, we hope to shed light on how a single molecule that is capable of exerting such divergent effects is also capable of promoting such distinct responses in different target tissues. J. Cell. Physiol. 224: 311–315, 2010.

Effects of leucine supplementation and resistance exercise on dexamethasone-induced muscle atrophy and insulin resistance in rats

OBJECTIVE We aimed to evaluate the effects of resistance exercise (RE) and leucine (LEU) supplementation on dexamethasone (DEXA)-induced muscle atrophy and insulin resistance. METHODS Male Wistar rats were randomly divided into DEXA (DEX), DEXA + RE (DEX-RE), DEXA + LEU (DEX-LEU), and DEXA + RE + LEU (DEX-RE-LEU) groups. Each group received DEXA 5 mg · kg(-1) · d(-1) for 7 d from drinking water and were pair-fed to the DEX group; LEU-supplemented groups received 0.135 g · kg(-1) · d(-1) through gavage for 7 d; the RE protocol was based on three sessions of squat-type exercise composed by three sets of 10 repetitions at 70% of maximal voluntary strength capacity. RESULTS The plantaris mass was significantly greater in both trained groups compared with the non-trained groups. Muscle cross-sectional area and fiber areas did not differ between groups. Both trained groups displayed significant increases in the number of intermediated fibers (IIa/IIx), a decreased number of fast-twitch fibers (IIb), an increased ratio of the proteins phospho(Ser2448)/total mammalian target of rapamycin and phospho(Thr389)/total 70-kDa ribosomal protein S6 kinase, and a decreased ratio of phospho(Ser253)/total Forkhead box protein-3a. Plasma glucose was significantly increased in the DEX-LEU group compared with the DEX group and RE significantly decreased hyperglycemia. The DEX-LEU group displayed decreased glucose transporter-4 translocation compared with the DEX group and RE restored this response. LEU supplementation worsened insulin sensitivity and did not attenuate muscle wasting in rats treated with DEXA. Conversely, RE modulated glucose homeostasis and fiber type transition in the plantaris muscle. CONCLUSION Resistance exercise but not LEU supplementation promoted fiber type transition and improved glucose homeostasis in DEXA-treated rats.

Distinct effects of leucine or a mixture of the branched-chain amino acids (leucine, isoleucine, and valine) supplementation on resistance to fatigue, and muscle and liver-glycogen degradation, in trained rats

OBJECTIVE The aim of this study was to evaluate the effects of the mixture of branched-chain amino acids (BCAAs) supplementation compared with leucine administered orally on muscle biochemical parameters of trained rats submitted to an exercise-induced protocol of glycogen depletion. METHODS After 6 wk of swimming exercise, 8 wk-old (250 g, adult) male Wistar rats were randomly divided into three experimental groups (n = 8 per group): the mixture of BCAAs (BCAAs), leucine (LEU), and placebo (PLA). All groups were submitted to swimming exercise for 6 wk and supplemented with either the mixture of BCAAs, leucine, or placebo during the last week of training. At week 7 of the protocol, the rats were submitted to an intermittent, progressive swimming test until exhaustion and sacrificed. Muscle gastrocnemius and liver were depicted to determine total glycogen, tricarboxylic acid cycle (TCA) intermediates, and enzymatic activities. Statistical evaluation was performed by one-way analysis of variance with Tukey post hoc test. RESULTS Both muscle and liver glycogen degradation ratio were significantly higher in the mixture of BCAAs group compared to the PLA group (P < 0.05) and the LEU group presented decreased liver glycogen degradation ratio compared with the mixture of BCAAs group (P < 0.05). Both muscle and liver glycogen content were significantly spared in the mixture of BCAAs and LEU groups compared to the PLA group (P < 0.01). A performance test demonstrated that LEU supplementation enhanced resistance to exhaustion compared to the mixture of BCAAs (P < 0.001), however, no difference was found when LEU supplementation was compared to PLA (P > 0.05) Muscle citrate content was significantly higher in the mixture of BCAAs group compared with the PLA group (P < 0.001). Muscle malate content was significantly elevated in the mixture of BCAAs group compared with both the PLA (P < 0.001) and LEU groups (P < 0.001). BCAT activity was significantly reduced in the mixture of BCAAs supplementation group compared with the LEU group (P < 0.001). CONCLUSION Leucine supplementation improved performance compared with the mixture of BCAAs supplementation, sparing muscle glycogen stores despite the augmentation of some TCA intermediate concentrations on the left side of the TCA cycle.

Potential therapeutic effects of branched-chain amino acids supplementation on resistance exercise-based muscle damage in humans

Branched-chain amino acids (BCAA) supplementation has been considered an interesting nutritional strategy to improve skeletal muscle protein turnover in several conditions. In this context, there is evidence that resistance exercise (RE)-derived biochemical markers of muscle soreness (creatine kinase (CK), aldolase, myoglobin), soreness, and functional strength may be modulated by BCAA supplementation in order to favor of muscle adaptation. However, few studies have investigated such effects in well-controlled conditions in humans. Therefore, the aim of this short report is to describe the potential therapeutic effects of BCAA supplementation on RE-based muscle damage in humans. The main point is that BCAA supplementation may decrease some biochemical markers related with muscle soreness but this does not necessarily reflect on muscle functionality.

Controversies of antioxidant vitamins supplementation in exercise: ergogenic or ergolytic effects in humans?

The aim of this commentary was to discuss the last studies regarding the effect of antioxidant vitamins supplementation on oxidative stress in exercise in humans. The inclusion criteria encompassed published studies done in adult males and females between 2006 and 2013. The keywords used in the search engine were: endurance athlete, diet, oxidative stress, physical activity, diet, nutrition, antioxidant, antioxidant status, vitamin C, vitamin A, vitamin E, β-carotene and combinations. Twelve studies were identified and organized according to the methodology and results of supplementation: ergogenic, ergolytic, partial or no difference between groups. The results of these studies showed no effect on physiological parameters and activity of antioxidant enzymes (n = 07), better response of the placebo treatment (ergolytic effect; n = 02), partial results (n = 01) and ergogenic results of antioxidant supplementation (n = 02). It is concluded that supplementation with antioxidant vitamins has controversial effects to oxidative damage induced by endurance exercise. The discordances among the studies are presented and discussed.

Comparative proteomic analysis of the aging soleus and extensor digitorum longus rat muscles using TMT labeling and mass spectrometry.

Sarcopenia describes an age-related decline in skeletal muscle mass, strength, and function that ultimately impairs metabolism and leads to poor balance, frequent falling, limited mobility, and a reduction in quality of life. Here we investigate the pathogenesis of sarcopenia through a proteomic shotgun approach. In brief, we employed tandem mass tags to quantitate and compare the protein profiles obtained from young versus old rat slow-twitch type of muscle (soleus) and a fast-twitch type of muscle (extensor digitorum longus, EDL). Our results disclose 3452 and 1848 proteins identified from soleus and EDL muscles samples, of which 78 and 174 were found to be differentially expressed, respectively. In general, most of the proteins were structural related and involved in energy metabolism, oxidative stress, detoxification, or transport. Aging affected soleus and EDL muscles differently, and several proteins were regulated in opposite ways. For example, pyruvate kinase had its expression and activity different in both soleus and EDL muscles. We were able to verify with existing literature many of our differentially expressed proteins as candidate aging biomarkers and, most importantly, disclose several new candidate biomarkers such as the glioblastoma amplified sequence, zero β-globin, and prolargin.

Dose and Latency Effects of Leucine Supplementation in Modulating Glucose Homeostasis: Opposite Effects in Healthy and Glucocorticoid-Induced Insulin-Resistance States

Dexamethasone (DEXA) is a potent immunosupressant and anti-inflammatory agent whose main side effects are muscle atrophy and insulin resistance in skeletal muscles. In this context, leucine supplementation may represent a way to limit the DEXA side effects. In this study, we have investigated the effects of a low and a high dose of leucine supplementation (via a bolus) on glucose homeostasis, muscle mass and muscle strength in energy-restricted and DEXA-treated rats. Since the leucine response may also be linked to the administration of this amino acid, we performed a second set of experiments with leucine given in bolus (via gavage) versus leucine given via drinking water. Leucine supplementation was found to produce positive effects (e.g., reduced insulin levels) only when administrated in low dosage, both via the bolus or via drinking water. However, under DEXA treatment, leucine administration was found to significantly influence this response, since leucine supplementation via drinking water clearly induced a diabetic state, whereas the same effect was not observed when supplied via the gavage.