Involvement of the Hsp70/TLR4/IL-6 and TNF-α pathways in delayed-onset muscle soreness.

Delayed-onset muscle soreness (DOMS) is a very common condition in athletes and individuals not accustomed to physical activity that occurs after moderate/high-intensity exercise sessions. The activation of microglial Toll-like receptor 4 (TLR4) in the spinal cord has been described to be important for the induction and maintenance of persistent pain. Based on that, we hypothesize that 70 kilodalton heat-shock protein (Hsp70), a mediator released by exercise, could activate microglial TLR4 in the spinal cord, releasing proinflammatory cytokines and contributing to the start of DOMS. In fact, we found that the knockout of TLR4, myeloid differentiation primary response 88 (MyD88), interleukin-6 (IL-6), or both tumor necrosis factor-α (TNF-α) receptor 1 and TNF-α receptor 2 in mice prevented the development of DOMS following acute aerobic exercise in contrast to the findings in male C57BL/6 wild-type mice. Furthermore, DOMS in exercised wild-type mice was also prevented after pre-treatment with microglia inhibitor, TLR4 antagonist, and anti-Hsp70 antibody. During exercise-induced DOMS, Hsp70 mRNA, TLR4 mRNA, and protein levels, as well as Iba-1 (a microglial marker), IL-6, and TNF-α protein levels, were increased in the muscle and/or spinal cord. Together, these findings suggest that Hsp70 released during exercise-induced DOMS activates the microglial TLR4/IL-6/TNF-α pathway in the spinal cord. Thus, the blockade of TLR4 activation may be a new strategy to prevent the development of DOMS before intense exercise.

Involvement of the Hsp70/TLR4/IL-6 and TNF-α pathways in delayed-onset muscle soreness

Delayed-onset muscle soreness (DOMS) is a very common condition in athletes and individuals not accustomed to physical activity that occurs after moderate/high-intensity exercise sessions. Activation of microglial Toll-like receptor 4 (TLR4) in the spinal cord has been described to be important for the induction and maintenance of persistent pain. Based on that, we hypothesize that 70 kilodalton heat shock protein (Hsp70), a mediator released by exercise, could activate microglial TLR4 in the spinal cord, releasing proinflammatory cytokines and contributing to the start of DOMS. In fact, we found that the knockout of TLR4, myeloid differentiation primary response 88 (MyD88), interleukin-6 (IL-6), or both TNF-a receptor 1 (TNFR1) and TNF-a receptor 2 (TNFR2) in mice prevented the development of DOMS following acute aerobic exercise in contrast to the findings in male C57BL/6 wild-type (WT) mice. Furthermore, DOMS in exercised WT mice was also prevented after pretreatment with microglia inhibitor, TLR4 antagonist and anti-Hsp70 antibody. During exercise-induced DOMS, Hsp70 mRNA, TLR4 mRNA and protein levels, as well as Iba-1 (a microglial marker), IL-6 and TNF-a protein levels, were increased in the muscle and/or spinal cord. Together, these findings suggest that Hsp70 released during exercise-induced DOMS activates the microglial TLR4/IL-6/TNF-a pathway in the spinal cord. Thus, the blockade of TLR4 activation may be a new strategy to prevent the development of DOMS before intense exercise.

Involvement of the Hsp70/TLR4/IL-6 and TNF-a pathways in delayed-onset muscle soreness

Delayed-onset muscle soreness (DOMS) is a very common condition in athletes and individuals not accustomed to physical activity that occurs after moderate/high-intensity exercise sessions. Activation of microglial Toll-like receptor 4 (TLR4) in the spinal cord has been described to be important for the induction and maintenance of persistent pain. Based on that, we hypothesize that 70 kilodalton heat shock protein (Hsp70), a mediator released by exercise, could activate microglial TLR4 in the spinal cord, releasing proinflammatory cytokines and contributing to the start of DOMS. In fact, we found that the knockout of TLR4, myeloid differentiation primary response 88 (MyD88), interleukin-6 (IL-6), or both TNF-a receptor 1 (TNFR1) and TNF-a receptor 2 (TNFR2) in mice prevented the development of DOMS following acute aerobic exercise in contrast to the findings in male C57BL/6 wild-type (WT) mice. Furthermore, DOMS in exercised WT mice was also prevented after pretreatment with microglia inhibitor, TLR4 antagonist and anti-Hsp70 antibody. During exercise-induced DOMS, Hsp70 mRNA, TLR4 mRNA and protein levels, as well as Iba-1 (a microglial marker), IL-6 and TNF-a protein levels, were increased in the muscle and/or spinal cord. Together, these findings suggest that Hsp70 released during exercise-induced DOMS activates the microglial TLR4/IL-6/TNF-a pathway in the spinal cord. Thus, the blockade of TLR4 activation may be a new strategy to prevent the development of DOMS before intense exercise.

Comparison of the Effects of Soy Protein and Whey Protein Supplementation during Exercise: a Systematic Review / Comparação dos Efeitos da Suplementação de Whey Protein e Proteína de Soja Durante o Exercício: uma Revisão Sistemática

The interest of the supplementation market for the soy protein consumption to optimize physical and metabolic performance after exercise is increasing. However, evidence suggests that the soy protein ingestion has lower anabolic properties when compared with whey protein. The purpose of this systematic review was to compare the effects of whey protein and soy protein supplementation on the muscle functions maintenance after exercise. This review was performed using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA). Articles were searched for in the Pubmed database and included studies comparing the effects of soy protein and whey protein consumption on protein synthesis, lean mass gain and oxidative stress reduction in response to endurance or resistance training. Thirteen trials were included in this review. The results showed that the whey protein consumption is superior to that of soy protein with respect to protein synthesis and lean mass gain, but soy protein showed superior results in reducing oxidative stress. Future research comparing both soy and whey protein are needed to define protein source to be used in nutritional interventions to protein synthesis, lean mass gain and oxidative stress in different populations. (AU)

O interesse do mercado de suplementação pelo consumo de proteína de soja para otimizar o desempenho físico e metabólico após o exercício está aumentando. No entanto, evidências sugerem que a ingestão da proteína de soja tem propriedades anabólicas mais baixas quando comparada à proteína do soro do leite. O objetivo desta revisão sistemática foi comparar os efeitos da suplementação com whey protein e proteína de soja na manutenção das funções musculares após o exercício. Esta revisão foi realizada usando os Itens de Relatório Preferidos para Revisões Sistemáticas e Meta-Análises (PRISMA). Os artigos foram pesquisados na base de dados Pubmed e incluíram estudos comparando os efeitos da proteína de soja e do consumo de proteínas do soro na síntese protéica, ganho de massa magra e redução do estresse oxidativo em resposta ao treinamento de resistência ou resistência. Treze ensaios foram incluídos nesta revisão. Os resultados mostraram que o consumo de proteína de soro é superior ao da proteína de soja em relação à síntese protéica e ao ganho de massa magra, mas a proteína de soja apresentou resultados superiores na redução do estresse oxidativo. Pesquisas futuras comparando a soja e a proteína do soro do leite são necessárias para definir a fonte protéica a ser usada em intervenções nutricionais para a síntese protéica, ganho de massa magra e estresse oxidativo em diferentes populações. (AU)

The expression of and preoperative correlation between heat-shock protein 70, euroscore, and lactate in patients undergoing cabg with cardiopulmonary bypass

Abstract Objetive: Coronary artery bypass grafting (CABG) with cardiopulmonary bypass (CPB) improved symptoms and increased survival and quality of life in patients with coronary artery disease. However, it should be the main cause of a complex organic systemic inflammatory response that greatly contributes to several postoperative adverse effects. Methods: We aimed to evaluate heat-shock protein 70 (HSP 70) expression as a morbimortality predictor in patients with preserved ventricular function undergoing coronary artery bypass grafting (CABG) with cardiopulmonary bypass (CPB) and to determine their association with the lactate as a marker of tissue hypoperfusion and the EuroSCORE risk score. This is a prospective, observational study including 46 patients and occurring between May and July 2016. Patients without ventricular dysfunction undergoing myocardial revascularization with extracorporeal circulation were included. They were divided into (1) complicated and (2) uncomplicated postoperative evolution groups. EuroSCORE, lactate levels, and HSP 70 expression and their correlations were determined. Results: Statistical analysis showed that the group with complicated evolution had higher EuroSCORE values than the other group. HSP 70 protein levels were significantly increased in the group with uncomplicated evolution and showed similar results. According to our results, HSP family proteins may be independent predictors of uncomplicated evolution in patients without ventricular dysfunction undergoing CABG with CPB. Conclusion: HSP 70 should be a good discriminator and protection marker for complications in cardiac surgery.

The Expression of and Preoperative Correlation between Heat-Shock Protein 70, EuroSCORE, and Lactate in Patients undergoing CABG with Cardiopulmonary Bypass.

OBJETIVE: Coronary artery bypass grafting (CABG) with cardiopulmonary bypass (CPB) improved symptoms and increased survival and quality of life in patients with coronary artery disease. However, it should be the main cause of a complex organic systemic inflammatory response that greatly contributes to several postoperative adverse effects. METHODS: We aimed to evaluate heat-shock protein 70 (HSP 70) expression as a morbimortality predictor in patients with preserved ventricular function undergoing coronary artery bypass grafting (CABG) with cardiopulmonary bypass (CPB) and to determine their association with the lactate as a marker of tissue hypoperfusion and the EuroSCORE risk score. This is a prospective, observational study including 46 patients and occurring between May and July 2016. Patients without ventricular dysfunction undergoing myocardial revascularization with extracorporeal circulation were included. They were divided into (1) complicated and (2) uncomplicated postoperative evolution groups. EuroSCORE, lactate levels, and HSP 70 expression and their correlations were determined. RESULTS: Statistical analysis showed that the group with complicated evolution had higher EuroSCORE values than the other group. HSP 70 protein levels were significantly increased in the group with uncomplicated evolution and showed similar results. According to our results, HSP family proteins may be independent predictors of uncomplicated evolution in patients without ventricular dysfunction undergoing CABG with CPB. CONCLUSION: HSP 70 should be a good discriminator and protection marker for complications in cardiac surgery.

Heat Shock Proteins: Protection and Potential Biomarkers for Ischemic Injury of Cardiomyocytes After Surgery.

The heat shock proteins are endogenous proteins with the ability to act as molecular chaperones. Methods that provide cell protection by way of some damage can positively influence the results of surgery. The present review summarizes current knowledge concerning the cardioprotective role of the heat shock proteins as occurs in heart damage, including relevant information about the stresses that regulate the expression of these proteins and their potential role as biomarkers of heart disease.

Heat shock proteins: protection and potential biomarkers for ischemic injury of cardiomyocytes after surgery

Abstract The heat shock proteins are endogenous proteins with the ability to act as molecular chaperones. Methods that provide cell protection by way of some damage can positively influence the results of surgery. The present review summarizes current knowledge concerning the cardioprotective role of the heat shock proteins as occurs in heart damage, including relevant information about the stresses that regulate the expression of these proteins and their potential role as biomarkers of heart disease.

Dietary Nutrients and Bioactive Substances Modulate Heat Shock Protein (HSP) Expression: A Review.

Interest in the heat shock proteins (HSPs), as a natural physiological toolkit of living organisms, has ranged from their chaperone function in nascent proteins to the remedial role following cell stress. As part of the defence system, HSPs guarantee cell tolerance against a variety of stressors, including exercise, oxidative stress, hyper and hypothermia, hyper and hypoxia and improper diets. For the past couple of decades, research on functional foods has revealed a number of substances likely to trigger cell protection through mechanisms that involve the induction of HSP expression. This review will summarize the occurrence of the most easily inducible HSPs and describe the effects of dietary proteins, peptides, amino acids, probiotics, high-fat diets and other food-derived substances reported to induce HSP response in animals and humans studies. Future research may clarify the mechanisms and explore the usefulness of this natural alternative of defense and the modulating mechanism of each substance.

Modulatory effects of arginine, glutamine and branched-chain amino acids on heat shock proteins, immunity and antioxidant response in exercised rats.

Heat shock proteins (HSPs) are endogenous proteins whose function is to maintain the cell's tolerance to insult, and glutamine supplementation is known to increase HSP expression during intense exercise. Since few studies have addressed the possibility that supplementation with other amino acids could have similar effects to that of glutamine, our objective was to evaluate the effects of leucine, valine, isoleucine and arginine as potential stimulators of HSPs 25, 60, 70 and 90 in rats subjected to acute exercise as a stressing factor. The immune markers, antioxidant system, blood parameters, glycogen and amino acid profile responses were also assessed. Male Wistar rats were divided into seven groups: control (rest, without gavage), vehicle (water), l-leucine, l-isoleucine, l-valine, l-arginine and l-glutamine. Except for the control, all animals were exercised and received every amino acid by oral gavage. Arginine supplementation up-regulated muscle HSP70 and HSP90 and serum HSP70, however, none of the amino acids affected the HSP25. All amino acids increased exercise-induced HSP60 expression, except for valine. Antioxidant enzymes were reduced by exercise, but both glutamine and arginine restored glutathione peroxidase, while isoleucine and valine restored superoxide dismutase. Exercise reduced monocyte, platelet, lymphocyte and erythrocyte levels, while leucine stimulated immune response, preserved the levels of the lymphocytes and increased leukocytes and maintained platelets at control levels. Plasma and muscle amino acid profiles showed specific metabolic features. The data suggest that the tissue-protecting effects of arginine could proceed by enhancing specific HSPs in the body.

Bioactivity of food peptides: biological response of rats to bovine milk whey peptides following acute exercise.

Background: Several physiologically beneficial effects of consuming a whey protein hydrolysate (WPH) have been attributed to the greater availability of bioactive peptides. Aims: The aim was to investigate the effect of four branched-chain amino acid- (BCAA-)containing dipeptides, present in WPH, on immune modulation, stimulation of HSP expression, muscle protein synthesis, glycogen content, satiety signals and the impact of these peptides on the plasma free amino acid profiles. Methods: The animals were divided in groups: control (rest, without gavage), vehicle (water), L-isoleucyl-L-leucine (lle-Leu), L-leucyl-L-isoleucine (Leu-lle), L-valyl-Lleucine (Val-Leu), L-leucyl-L-valine (Leu-Val) and WPH. All animals were submitted to acute exercise, except for control. Results: lle-Leu stimulated immune response, hepatic and muscle glycogen and HSP60 expression, whereas Leu-Val enhanced HSP90 expression. All dipeptides reduced glucagon-like peptide-1 and glucose-dependent insulinotropic polypeptide, no changes were observed on leptin. All peptides inhibited NF-kB expression. The plasma amino acid time-course showed peptide-specific and isomer-specific metabolic features, including increases of the BCAAs. Conclusion: The data indicate that lle-Leu was effective to attenuate immune-suppression exercise-induced, promoted glycogen content and stimulated anti-stress effect (HSP). Furthermore, Leu-Val increased HSP90, p-4EBP1, p-mTOR and p-AMPK expression. The data suggest the involvement of these peptides in various beneficial functions of WPH consumption.

Consumption of açai (Euterpe oleracea Mart.) functional beverage reduces muscle stress and improves effort tolerance in elite athletes: a randomized controlled intervention study.

The study analyzed the effect of an açai (Euterpe oleracea Mart.) functional beverage (AB) on muscle and oxidative stress markers, cardiorespiratory responses, perceived exertion, and time-to-exhaustion during maximal treadmill running. The beverage was developed as an ergogenic aid for athletes and contained 27.6 mg of anthocyanins per dose. Fourteen athletes performed 3 exercise tests: a ramp-incremental maximal exercise test and 2 maximal exercise bouts performed in 2 conditions (AB and without AB (control)) at 90% maximal oxygen uptake. Blood was collected at baseline and after maximal exercise in both conditions to determine biomarkers. AB increased time to exhaustion during short-term high-intensity exercise (mean difference: 69 s, 95% confidence interval = -296 s to 159 s, t = 2.2, p = 0.045), attenuating the metabolic stress induced by exercise (p < 0.05). AB also reduced perceived exertion and enhanced cardiorespiratory responses (p < 0.05). The AB may be a useful and practical ergogenic aid to enhance performance during high-intensity training.

Chia (Salvia hispanica L.) enhances HSP, PGC-1α expressions and improves glucose tolerance in diet-induced obese rats.

OBJECTIVE: The aim of this study was to investigate the effects of chia seed and chia oil on heat shock protein (HSP) and related parameters in diet-induced obese rats. METHODS: Animals were divided in six groups: control, high-fat and high-fructose diet (HFF), and HFF with chia seed or chia oil in short (6-wk) and long (12-wk) treatments. Plasma indicators of glucose tolerance and liver damage, skeletal muscle expression of antioxidant enzymes, and proteins controlling oxidative energy metabolism were determined. The limit of significance was set at P < 0.05. RESULTS: The HFF diet induced glucose intolerance, insulin resistance, oxidative stress, and altered parameters related to obesity complications. The consumption of chia seed or chia oil did not reduce body weight gain or abdominal fat accumulation. However, chia seed and chia oil in both treatments improved glucose and insulin tolerance. Chia oil in both treatments induced expression of HSP70 and HSP25 in skeletal muscle. Short treatment with chia seed increased expression of HSP70, but not HSP25. Chia oil in both treatments restored superoxide dismutase and glutathione peroxidase expression. Extended treatment with chia seed and short treatment with chia oil restored peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) expression. CONCLUSION: Chia oil restored the antioxidant system and induced the expression of a higher number of proteins than chia seed. The present study demonstrated new properties and molecular mechanisms associated with the beneficial effects of chia seed and chia oil consumption in diet-induced obese rats.

Dietary supplement usage and motivation in Brazilian road runners.

BACKGROUND: The consumption of dietary supplements is highest among athletes and it can represent potential a health risk for consumers. OBJECTIVE: The aim of this study was to determine the prevalence of consumption of dietary supplements by road runners. METHODS: We interviewed 817 volunteers from four road races in the Brazilian running calendar. The sample consisted of 671 male and 146 female runners with a mean age of 37.9 ± 12.4 years. RESULTS: Of the sample, 28.33% reported having used some type of dietary supplement. The main motivation for this consumption is to increase in stamina and improve performance. The probability of consuming dietary supplements increased 4.67 times when the runners were guided by coaches. The consumption of supplements was strongly correlated (r = 0.97) with weekly running distance, and also highly correlated (r = 0.86) with the number of years the sport had been practiced. The longer the runner had practiced the sport, the higher the training volume and the greater the intake of supplements. The five most frequently cited reasons for consumption were: energy enhancement (29.5%), performance improvement (17.1%), increased level of endurance (10.3%), nutrient replacement (11.1%), and avoidance of fatigue (10.3%). About 30% of the consumers declared more than one reason for taking dietary supplements. The most consumed supplements were: carbohydrates (52.17%), vitamins (28.70%), and proteins (13.48%). CONCLUSIONS: Supplement consumption by road runners in Brazil appeared to be guided by the energy boosting properties of the supplement, the influence of coaches, and the experience of the user. The amount of supplement intake seemed to be lower among road runners than for athletes of other sports. We recommend that coaches and nutritionists emphasise that a balanced diet can meet the needs of physically active people.

Whey protein hydrolysate enhances HSP90 but does not alter HSP60 and HSP25 in skeletal muscle of rats.

Whey protein hydrolysate (WPH) intake has shown to increase HSP70 expression. The aim of the present study was to investigate whether WPH intake would also influences HSP90, HSP60 and HSP25 expression, as well as associated parameters. Forty-eight male Wistar rats were divided into sedentary (unstressed) and exercised (stressed) groups, and were fed with three different sources of protein: whey protein (WP), whey protein hydrolysate (WPH) and casein (CAS) as a control, based on the AIN93G diet for 3 weeks. WPH intake increased HSP90 expression in both sedentary and exercised animals compared to WP or CAS, however no alteration was found from exercise or diet to HSP60 or HSP25. Co-chaperone Aha1 and p-HSF1 were also increased in the exercised animals fed with WPH in comparison with WP or CAS, consistent with enhanced HSP90 expression. VEGF and p-AKT were increased in the WPH exercised group. No alteration was found in BCKDH, PI3-Kinase (p85), GFAT, OGT or PGC for diet or exercise. The antioxidant system GPx, catalase and SOD showed different responses to diet and exercise. The data indicate that WPH intake enhanced factors related to cell survival, such as HSP90 and VEGF, but does not alter HSP60 or HSP25 in rat skeletal muscle.

Whey protein hydrolysate increases translocation of GLUT-4 to the plasma membrane independent of insulin in wistar rats.

Whey protein (WP) and whey protein hydrolysate (WPH) have the recognized capacity to increase glycogen stores. The objective of this study was to verify if consuming WP and WPH could also increase the concentration of the glucose transporters GLUT-1 and GLUT-4 in the plasma membrane (PM) of the muscle cells of sedentary and exercised animals. Forty-eight Wistar rats were divided into 6 groups (n = 8 per group), were treated and fed with experimental diets for 9 days as follows: a) control casein (CAS); b) WP; c) WPH; d) CAS exercised; e) WP exercised; and f) WPH exercised. After the experimental period, the animals were sacrificed, muscle GLUT-1 and GLUT-4, p85, Akt and phosphorylated Akt were analyzed by western blotting, and the glycogen, blood amino acids, insulin levels and biochemical health indicators were analyzed using standard methods. Consumption of WPH significantly increased the concentrations of GLUT-4 in the PM and glycogen, whereas the GLUT-1 and insulin levels and the health indicators showed no alterations. The physical exercise associated with consumption of WPH had favorable effects on glucose transport into muscle. These results should encourage new studies dealing with the potential of both WP and WPH for the treatment or prevention of type II diabetes, a disease in which there is reduced translocation of GLUT-4 to the plasma membrane.

Whey protein hydrolysate enhances the exercise-induced heat shock protein (HSP70) response in rats.

Whey protein has been suggested to be potential protective agent against various forms of stress. The heat shock protein HSP70 confers greater cellular tolerance against stressors. The present study evaluated the effects of whey protein intake on HSP70 expression. Forty-eight male Wistar rats were divided into sedentary and exercised groups, and each group was fed as a protein source casein (CAS), whey protein (WP) or whey protein hydrolysate (WPH) for 3weeks. Exercise on a treadmill was used as the source of stress in the animals from the exercised group. The results showed a larger increase in HSP70 expression in the soleus, gastrocnemius and lung of the WPH-fed rats than WP or casein-fed rats. HSP70 expression in the sedentary rats was very low, independent of the diet or tissue. Protein carbonyls were lower in the group that consumed WPH. These data suggest that the consumption of WPH enhances HSP70 expression.

Physiological and physical effects of different milk protein supplements in elite soccer players.

Brazilian soccer championships involve a large number of teams and are known to cause stress and loss of muscle mass besides other negative physical consequences. This study was designed to compare the effects produced by three types of protein supplements on body composition, biochemical parameters and performance of a top Brazilian professional soccer team during an actual tournament. Twenty-four athletes assessed as having a normal nutrient intake were divided into three groups according to supplementation. Immediately after each daily training, the athletes received 1 g × kg(-1) of body weight × day(-1) of either whey protein (WP), hydrolyzed whey protein (HWP) or casein (CAS) for eight weeks. Before and after the experimental period, anthropometric characteristics, physical performance by the yo-yo and 3000m tests, and several biochemical variables in blood (uric acid, total cholesterol, HDL-cholesterol, creatinine, glucose) were measured. While no improvement in physical performance was observed with regard to the applied treatments, casein supplementation resulted in muscle mass increase (p<0.039), while WP and HWP favoured the maintenance of the initial muscle mass. Moreover, the eight-week intervention was found to cause no abnormalities in biochemical and anthropometric variables monitored, but instead, the intervention showed to be positive in comparison to the adverse anthropometric changes, when no supplementation was made. It was concluded that supplementation immediately after training sessions with any of the three sources of protein during the competitive period is beneficial and safe, as well as capable of sustaining or even increasing muscle mass.