RESUMEN
BACKGROUND & AIMS: Adequate protein intake is crucial to maintain body protein content in elderly subjects, but quality of dietary proteins should be also considered since amino acid composition and rate of protein digestion modulate amino acid availability. This study investigates whether the efficacy of optimal protein intake levels for protein retention in the elderly is influenced by protein quality. METHODS: We investigated the effect of a 10-day adequate-protein (AP) or high-protein (HP) diet together with the protein source as caseins (CAS) or soluble milk proteins (PRO) on whole-body (WB) protein synthesis (PS) and protein breakdown (PB) in 4 groups of healthy elderly men (mean ± SEM: 71.8 ± 24.4 yr). The study consisted of two periods of 4 h each: a post-absorptive period and a postprandial period. The fed state was defined by consumption every 20 min and for 4 h, of either 15 g or 30 g of PRO or CAS. Steady-state WB and splanchnic leucine kinetics were measured using a continuous infusion of L-[1-13C]leucine in the postabsorptive state and L-[1-13C]leucine infusion plus oral L-[5,5,5-2H3]leucine in the postprandial state. RESULTS: WB PS was stimulated by feeding only with HP diets, whereas WB PB corrected for splanchnic extraction showed a similar pattern of post-feeding decrease in all groups. Consequently, net leucine balance was greater in the postprandial state after HP meals than after AP meals, with PRO meals leading to a better postprandial leucine balance (3.63 ± 0.16 µmol kg FFM(-1) min(-1)vs. 2.77 ± 0.21 µmol kg FFM(-1) min(-1) for PRO HP and CAS HP, respectively; P = 0.005). CONCLUSION: Postprandial protein retention was better improved in elderly men by an increase in protein intake when the protein supplementation was provided as fast-digesting proteins that induce high leucine availability.
Asunto(s)
Aminoácidos Esenciales/administración & dosificación , Proteínas en la Dieta/administración & dosificación , Estado Nutricional , Anciano , Aminoácidos Esenciales/sangre , Índice de Masa Corporal , Peso Corporal , Caseínas/administración & dosificación , Productos Lácteos/análisis , Método Doble Ciego , Humanos , Insulina/sangre , Leucina/administración & dosificación , Leucina/farmacocinética , Masculino , Comidas , Proteínas de la Leche/administración & dosificación , Periodo PosprandialRESUMEN
Epidemiological studies suggest that chronic consumption of trans MUFA may alter muscle insulin sensitivity. The major sources of dietary trans MUFA (dairy fat vs. industrially hydrogenated oils) have different isomeric profiles and thus probably different metabolic consequences. These effects may involve alterations in muscle mitochondrial oxidative capacity, which may in turn promote insulin resistance if fatty acid oxidation is reduced. We report that in Wistar rats, an 8 week diet enriched (4% of energy intake) in either dairy, industrial, or control MUFA did not alter insulin and glucose responses to an intraperitoneal glucose tolerance test (1g/kg). In C2C12 myotubes, vaccenic and elaidic acids did not modify insulin sensitivity compared with oleic acid. Furthermore, the ex vivo total, mitochondrial and peroxisomal oxidation rates of [1-(14)C]oleic, vaccenic, and elaidic acids were similar in soleus and tibialis anterior rat muscle. Finally, an 8 week diet enriched in either dairy or industrial trans MUFA did not alter mitochondrial oxidative capacity in these two muscles compared with control MUFA but did induce a specific reduction in soleus mitochondrial ATP and superoxide anion production (P<0.01 vs. control). In conclusion, dietary trans MUFA of dairy or industrial origin have similar effects and do not impair muscle mitochondrial capacity and insulin sensitivity.
Asunto(s)
Grasas de la Dieta/farmacología , Industrias , Insulina/farmacología , Mitocondrias/efectos de los fármacos , Músculos/efectos de los fármacos , Aceites/farmacología , Ácidos Grasos trans/farmacología , Adenosina Trifosfato/biosíntesis , Alimentación Animal , Animales , Composición Corporal , Línea Celular , Grasas de la Dieta/metabolismo , Masculino , Ratones , Mitocondrias/metabolismo , Membranas Mitocondriales/metabolismo , Músculos/metabolismo , Aceites/metabolismo , Oxidación-Reducción , Ratas , Ratas Wistar , Superóxidos/metabolismo , Ácidos Grasos trans/metabolismoRESUMEN
OBJECTIVE: Obesity and insulin resistance are associated with muscle mitochondrial dysfunction, which might be related to impairment of mitochondrial protein synthesis. This study aimed at investigating mixed and mitochondrial protein synthesis in skeletal muscle in response to dietary manipulations. RESEARCH METHODS AND PROCEDURES: High-sucrose (SU) and high-fat, high-sucrose (F) diets were provided for 6 weeks to Wistar rats at standard (N) and high (H) energy intakes and compared with controls. Fractional synthesis rates of mixed (FSRPT) and mitochondrial (FSRm) proteins within the oxidative (soleus) and glycolytic (tibialis) muscles were measured using stable isotope flooding dose technique using L-[13C]-valine. Carbonyl content, citrate synthase, and cytochrome c oxidase activities were assayed spectrophotometrically on isolated mitochondria. RESULTS: In the soleus, FSRPT was increased by 40% in the NSU and NF groups and by 65% in the HSU and HF groups (p<0.001 vs. control). FSRm was increased with high-fat diets (NF, +16%; HF, +32%; p<0.01). In the tibialis, FSR(PT) was enhanced in all experimental groups (+31% to 37%, p<0.05 vs. control). FSRm was augmented in the NSU, NF, and HF groups (+28% to 32%, p<0.01). Cytochrome c oxidase activity was significantly decreased in all experimental groups in the soleus (p<0.001). DISCUSSION: Muscle mixed and mitochondrial protein FSR are enhanced after short-term dietary intervention known to induce insulin resistance and obesity. Adaptations are muscle type specific and may not explain alterations in mitochondrial oxidative capacity but might contribute to maintain mitochondrial functioning.
Asunto(s)
Grasas de la Dieta/metabolismo , Mitocondrias/metabolismo , Proteínas Mitocondriales/metabolismo , Músculos/patología , Sacarosa/metabolismo , Alimentación Animal , Animales , Complejo IV de Transporte de Electrones/metabolismo , Ingestión de Energía , Resistencia a la Insulina , Masculino , Músculo Esquelético/metabolismo , Obesidad/metabolismo , Ratas , Ratas WistarRESUMEN
OBJECTIVE: Mitochondrial dysfunction might predispose individuals to develop insulin resistance. Our objective was to determine whether mitochondrial dysfunction or insulin resistance was the primary event during high-fat (HF) diet. RESEARCH METHODS AND PROCEDURES: Rats were fed an HF diet for 0, 3, 6, 9, 14, 20, or 40 days and compared with control. Soleus and tibialis muscle mitochondrial activity were assessed using permeabilized fiber technique. Insulin [area under the curve for insulin (AUC(I))] and glucose [area under the curve for glucose (AUC(G))] responses to intraperitoneal glucose tolerance test as well as fasting plasma non-esterified fatty acids (NEFAs), triglyceride, and glycerol concentrations were determined. RESULTS: AUC(I) and AUC(G) were altered from Day 6 (p < 0.01 vs. Day 0). In soleus, oxidative phosphorylation (OXPHOS) activity was transiently enhanced by 26% after 14 days of HF diet (p < 0.05 vs. Day 0) conjointly with 62% increase in NEFA concentration (p < 0.05 vs. Day 0). This was associated with normalized AUC(G) at Day 14 and with a decline of plasma NEFA concentration together with stabilization of intra-abdominal adiposity at Day 20. Prolongation of HF diet again caused an increase in plasma NEFA concentration, intra-abdominal adiposity, AUC(I), and AUC(G). At Day 40, significant decrease in OXPHOS activity was observed in soleus. DISCUSSION: Mitochondria first adapt to overfeeding in oxidative muscle limiting excess fat deposition. This potentially contributes to maintain glucose homeostasis. Persistent overfeeding causes insulin resistance and results in a slow decline in oxidative muscle OXPHOS activity. This shows that the involvement of mitochondria in the predisposition to insulin resistance is mainly due to an inability to face prolonged excess fat delivery.
Asunto(s)
Dieta , Grasas de la Dieta/administración & dosificación , Resistencia a la Insulina , Metabolismo de los Lípidos/efectos de los fármacos , Mitocondrias Musculares/fisiología , Obesidad/metabolismo , Adaptación Fisiológica , Tejido Adiposo/metabolismo , Animales , Área Bajo la Curva , Glucemia/metabolismo , Ácidos Grasos no Esterificados/sangre , Prueba de Tolerancia a la Glucosa , Glicerol/metabolismo , Insulina/sangre , Masculino , Mitocondrias Musculares/metabolismo , Obesidad/fisiopatología , Distribución Aleatoria , Ratas , Ratas Wistar , Factores de Tiempo , Triglicéridos/metabolismoRESUMEN
Caloric restriction (CR) delays the onset of age-related mitochondrial abnormalities but does not prevent the decline in ATP production needed to sustain muscle protein fractional synthesis rate (FSR) and contractile activity. We hypothesized that improving mitochondrial activity and FSR using a CR diet with maintained protein intakes could enhance myofibrillar protein FSR and consequently improve muscle strength in aging rats. Wistar rats (21 months old) were fed either an ad libitum (AL), 40% protein-energy restricted (PER) or 40% AL-isonitrogenous energy restricted (ER) diet for 5 months. ATP production, electron transport chain activity, reactive oxygen species (ROS) generation, protein carbonyl content and FSR were determined in both tibialis anterior (TA) and soleus muscle mitochondria. Myosin and actin FSR and grip force were also investigated. The ER diet led to improved mitochondrial activity and ATP production in the TA and soleus muscles in comparison with PER. Furthermore, mitochondrial FSR in the TA was enhanced under the ER diet but diminished under the PER. Mitochondrial protein carbonyl content was decreased by both the ER and PER diets. The ER diet was able to improve myosin and actin FSR and grip force. Therefore, the synergistic effects of CR with maintained protein intake may help to limit the progression of sarcopenia by optimizing the turnover rates and functions of major proteins in skeletal muscle.
Asunto(s)
Restricción Calórica , Proteínas en la Dieta/metabolismo , Mitocondrias Musculares/metabolismo , Músculo Esquelético/metabolismo , Adenosina Trifosfato/biosíntesis , Envejecimiento , Animales , Masculino , Miosinas/metabolismo , Estrés Oxidativo , Oxígeno/metabolismo , Fósforo/metabolismo , Ratas , Ratas Wistar , Superóxidos/metabolismoRESUMEN
Obesity is often associated with insulin resistance and mitochondrial dysfunction within skeletal muscles, but the causative factors are not clearly identified. The present study examined the role of nutrition, both qualitatively and quantitatively, in the induction of muscle mitochondrial defects. Two experimental diets [high sucrose (SU) and high fat (F)] were provided for 6 wk to male Wistar rats at 2 levels of energy [standard (N) and high (H)] and compared with a standard energy cornstarch-based diet (C). Insulin sensitivity (intraperitoneal glucose tolerance test, IPGTT) and intramyocellular triglyceride (IMTG) content (1H MRS) were determined at wk 5. Mitochondrial oxidative phosphorylation and superoxide anion radical (MSR) production were assessed on soleus (oxidative) and tibialis (glycolytic) muscles. Experimental diets induced hyperinsulinemia during IPGTT (P < 0.01 vs. C). Rats in the HSU and HF groups were hyperglycemic relative to the C group, P < 0.05 vs. C. The severity of insulin resistance paralleled IMTG accumulation (P < 0.05). In soleus, mitochondrial respiration and ATP production rates were lower in HSU and HF than in C (P < 0.05). By contrast, respiration was unaffected by the diets in tibialis, whereas ATP production tended to be lower in rats fed the experimental diets compared with C (P = 0.09). Mitochondrial adaptations were associated with more than a 50% reduction in MSR production in HSU and HF compared with C in both soleus (P < 0.05) and tibialis (P < 0.01). Changes in mitochondrial functions in the NSU and NF groups were intermediate and not significantly different from C. Therefore, excess fat or sucrose and more importantly, excess energy intake by rats is associated with muscle type-specific mitochondrial adaptations, which contribute to decrease mitochondrial production of ATP and reactive oxygen species.
Asunto(s)
Adenosina Trifosfato/biosíntesis , Carbohidratos de la Dieta/farmacología , Grasas de la Dieta/farmacología , Mitocondrias Musculares/efectos de los fármacos , Músculo Esquelético/efectos de los fármacos , Fosforilación Oxidativa/efectos de los fármacos , Animales , Carbohidratos de la Dieta/administración & dosificación , Grasas de la Dieta/administración & dosificación , Ingestión de Energía , Resistencia a la Insulina , Masculino , Mitocondrias Musculares/metabolismo , Mitocondrias Musculares/fisiología , Músculo Esquelético/metabolismo , Ratas , Ratas Wistar , Superóxidos/metabolismoRESUMEN
Immune response is dependent on synthesis and secretion of various proteins. Hence, in vivo determination of protein synthesis rate in leukocytes may reflect the functional activity of these cells under conditions of immune deficiency such as aging. Because (i) insulin stimulates protein turnover in various tissues and (ii) elderly people often suffer from a decrease in insulin sensitivity, we sought to determine the effect of physiological hyperinsulinemia (0.7mU/kg fat free mass/min) on protein synthesis rate in peripheral blood mononuclear cells (PBMCs) and polymorphonuclear neutrophils (PMNs) from 8 adult (24+/-6yr) and 9 elderly (69+/-4yr) healthy subjects under strict euglycemia (0.9g/l). The body mass index was significantly higher in elderly volunteers compared to the younger ones (P<0.05) whereas insulin sensitivity was lower as shown by either the M value (P<0.05) or the glucose disposal rate (GDR)/insulinemia ratio (P<0.05). The density of the insulin receptor at PBMCs and PMNs surfaces was not altered by either age or insulin infusion. In comparison with the baseline values, insulin highly stimulated the fractional synthesis rate (FSR) of mixed PBMCs (4.12+/-0.56%/d vs. 6.94+/-0.70%/d, P<0.05) and PMNs (1.05+/-0.28%/d vs. 2.44+/-0.41%/d, P<0.05) proteins. No age effect was observed either during the baseline (PBMCs: 4.31+/-0.30%/d, PMNs: 1.55+/-0.28%/d) period or after insulin infusion (PBMCs: 6.86+/-0.69%/d, PMNs: 2.76+/-0.47%/d). We conclude that insulin is able to stimulate protein synthesis in leukocyte populations independently of the age of the subjects.
Asunto(s)
Envejecimiento/inmunología , Envejecimiento/metabolismo , Insulina/fisiología , Leucocitos/metabolismo , Biosíntesis de Proteínas , Adulto , Anciano , Aminoácidos/sangre , Glucemia/metabolismo , Índice de Masa Corporal , Femenino , Técnica de Clampeo de la Glucosa , Humanos , Insulina/sangre , Insulina/metabolismo , Recuento de Leucocitos , Leucocitos Mononucleares/metabolismo , Masculino , Neutrófilos/metabolismoRESUMEN
It is well known that the intestinal availability of Ca from Ca-rich mineral waters is equivalent to that of milk Ca. However, the effect of associated anions on Ca urinary loss needs to be addressed. The aim of the current study was to compare, under ordinary conditions of consumption, milk and a SO4-rich mineral water as the Ca provider in a large number of subjects consuming the same quantity of Ca from the two sources in a crossover study lasting for an extended period. Thirty-seven healthy women completed a 12-week protocol, divided into four periods of 3 weeks (W). In the first (W1-3) and third (W6-9) periods, dietary Ca intake was restricted to 600 mg/d. In the second (W4-6) and final (W10-12) periods, either 400 ml/d medium-fat milk or 1 litre of a Ca- and SO4-rich mineral water, each providing about 480 mg Ca/d, was added to the diet in a random manner. Dietary evaluation, blood and urinary measures were performed during the last week (W6 and W12) of each Ca supplementation period. The urinary excretion of Ca was higher (0.5 mmol/d more) with water than with milk (P<0.001). An examination of all the dietary factors known to influence calciuria suggested that the acidogenic action of SO4 was responsible for this increased calciuria. Thus, despite an equal Ca intake and assuming an unchanged intestinal absorption, these results suggest that Ca balance is better with milk consumption than with CaSO4-rich water.
Asunto(s)
Sulfato de Calcio/administración & dosificación , Calcio/orina , Leche , Aguas Minerales/administración & dosificación , Adulto , Aminoácidos/análisis , Animales , Biomarcadores/análisis , Creatinina/orina , Estudios Cruzados , Suplementos Dietéticos , Ingestión de Líquidos , Femenino , Humanos , Magnesio/orina , Sodio/orinaRESUMEN
Responses of whole body glucose disposal (GDR) and protein breakdown (PB) to physiological insulin levels are altered in nondiabetic elderly subjects. Amino acids enhance inhibition of PB by insulin in young subjects. We hypothesized that addition of amino acid to insulin may improve the defect in PB regulation by insulin in elderly people. Therefore, we investigated the effect of hyperinsulinemia combined to either euaminoacidemia (EuAA) or hyperaminoacidemia (HyperAA) on GDR and PB, using isotopic dilution of D-[6,6-2H2]glucose and L-[1-13C]leucine, in young (mean +/- SEM, 24.4 +/- 0.8 yr) and elderly (70.2 +/- 0.7 yr) subjects. GDR was lower in elderly than in young subjects in all situations (P < 0.05). Despite a greater inhibition with HyperAA, PB was less inhibited in elderly than in young subjects during both clamps (ratio between change over basal PB and change over basal insulinemia, -0.014 +/- 0.002 vs. -0.024 +/- 0.003 in EuAA and -0.022 +/- 0.002 vs. -0.036 +/- 0.003 micromol/ml.microU/kg fat-free mass.min in HyperAA; elderly vs. young, P < 0.05). In conclusion, in nondiabetic elderly subjects, PB is less inhibited by insulin with either basal or high amino acid concentrations. Addition of amino acid potentiates insulin-induced suppression of PB in both groups to the same extent, suggesting a specific dysregulation of PB by insulin with age.
Asunto(s)
Envejecimiento/metabolismo , Aminoácidos/farmacología , Insulina/farmacología , Proteínas/metabolismo , Adulto , Anciano , Envejecimiento/sangre , Aminoácidos/sangre , Glucemia/metabolismo , Composición Corporal , Dióxido de Carbono , Espiración , Glucosa/metabolismo , Humanos , Insulina/sangre , Leucina/sangre , Leucina/metabolismo , Masculino , Concentración Osmolar , Ácidos Pentanoicos/sangre , Proteínas/antagonistas & inhibidoresRESUMEN
Leucine has a major anabolic impact on muscle protein synthesis in young as in old animals. However, myosin heavy chain (MHC), sarcoplasmic and mitochondrial proteins may differently respond to anabolic factors, especially during aging. To test this hypothesis, fractional synthesis rates (FSR) of the three muscle protein fractions were measured using a flooding dose of [1-(13)C] phenylalanine, in gastrocnemius muscle of adult (8 months) and old (22 months) rats, either in postabsorptive state (PA), or 90-120 min after ingestion of a alanine-supplemented meal (PP+A) or a leucine-supplemented meal (PP+L). In adult and old rats, in comparison with PA, leucine stimulated mitochondrial (adult: 0.260+/-0.011 vs 0.238+/-0.012%h(-1); old: 0.289+/-0.010 vs 0.250+/-0.010%h(-1); PP+L vs PA, P<0.05) and sarcoplasmic (adult: 0.182+/-0.011 vs 0.143+/-0.006%h(-1); old: 0.195+/-0.010 vs 0.149+/-0.008%h(-1); PP+L vs PA, P<0.05) protein FSR, but not MHC synthesis in old rats (0.101+/-0.009 vs 0.137+/-0.018%h(-1); PP+L vs PA, P=NS). In conclusion, synthesis of specific muscle protein is activated by leucine supplementation, but MHC may be less sensitive to anabolic factors with aging.
Asunto(s)
Envejecimiento/fisiología , Leucina/administración & dosificación , Proteínas Mitocondriales/biosíntesis , Proteínas Musculares/biosíntesis , Músculo Esquelético/metabolismo , Alanina/administración & dosificación , Animales , Dieta , Masculino , Cadenas Pesadas de Miosina/biosíntesis , Fenilalanina/administración & dosificación , Ratas , Ratas Wistar , Retículo Sarcoplasmático/metabolismoRESUMEN
Immune cell functions can be evaluated in vivo by measuring their specific protein fractional synthesis rates (FSR). Using stable isotope dilution techniques, we describe a new method allowing simultaneous in vivo assessment of FSR in two leukocyte populations in healthy human subjects, using small blood samples. Peripheral blood mononuclear cell (PBMC) and polymorphonuclear neutrophil (PMN) FSR were measured during primed continuous intravenous infusion of l-[1-(13)C]leucine. Immune cells from 6 ml of whole blood were isolated by density gradient centrifugation. In a first study, we calculated the FSR using plasma [(13)C]leucine or alpha-[(13)C]ketoisocaproate (KIC) enrichments as precursor pools. In a second study, we compared protein FSR in leukocytes, using enrichments of either intracellular or plasma free [(13)C]leucine as immediate precursor pools. The present approach showed a steady-state enrichment of plasma and circulating immune cell free [(13)C]leucine precursor pools. The linearity of labeled amino acid incorporation rate within mixed PBMC and PMN proteins also was verified. Postabsorptive protein FSR was 4.09 +/- 0.39%/day in PBMC and 1.44 +/- 0.08%/day in PMN when plasma [(13)C]KIC was the precursor pool. The difference between PBMC and PMN FSR was statistically significant, whatever the precursor pool used, suggesting large differences in their synthetic activities and functions. Use of the plasma [(13)C]KIC pool led to an underestimation of leukocyte FSR compared with the intracellular pool (PBMC: 6.04 +/- 0.94%/day; PMN: 2.98 +/- 0.30%/day). Hence, the intracellular free amino acid pool must be used as precursor to obtain reliable results. In conclusion, it is possible to assess immune cell metabolism in vivo in humans by using small blood samples to directly indicate their metabolic activity in various clinical situations and in response to regulating factors.
Asunto(s)
Histocitoquímica/métodos , Leucina/farmacocinética , Leucocitos Mononucleares/metabolismo , Neutrófilos/metabolismo , Biosíntesis de Proteínas , Adulto , Isótopos de Carbono , Separación Celular , Humanos , Líquido Intracelular/metabolismo , Cetoácidos/sangre , Cetoácidos/metabolismo , Leucina/sangre , Leucina/metabolismo , Plasma/metabolismo , Valores de Referencia , Reproducibilidad de los Resultados , Factores de TiempoRESUMEN
PURPOSE: The purpose of this study was to determine whether an ultra-endurance event is a strong stimulus to increase albumin synthesis involved in the process of intravascular albumin mass increase associated with transient hypervolemia. METHODS: The albumin synthetic rate was measured in six young men, 3 d before (C) and on the 1st (R1) and 8th (R8) days of the recovery from an ultra-endurance trial (5 h daily for 4 d). Albumin fractional (FSR) and absolute (ASR) synthetic rate were determined using a primed-constant infusion of [1(-13) C] leucine. Plasma volume (PV) using Evans Blue dye dilution and total body water (TBW) using bioelectrical impedance analysis were measured on C, R1, and R8. RESULTS: On R1 as compared with C: 1). PV (+23.3 +/- 3.2%; PAsunto(s)
Albúminas/biosíntesis
, Resistencia Física/fisiología
, Adulto
, Agua Corporal
, Peso Corporal
, Impedancia Eléctrica
, Humanos
, Masculino
, Volumen Plasmático