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Certain conditions, such as several weeks of high-fat diet, disrupt endoplasmic reticulum (ER) homeostasis and activate an adaptive pathway referred as the unfolded protein response. When the unfolded protein response fails, the result is the development of inflammation and insulin resistance. These two pathological states are known to be improved by regular exercise training but the mechanisms remain largely undetermined. As it has recently been shown that the unfolded protein response is regulated by exercise, we hypothesised that concomitant treadmill exercise training (HFD+ex) prevents ER homeostasis disruption and its downstream consequences induced by a 6-week high-fat diet (HFD) in mice by activating the protective unfolded protein response. Several well-documented markers of the unfolded protein response were measured in the soleus and tibialis anterior muscles as well as in the liver and pancreas. In HFD mice, an increase in these markers was observed (from 2- to 15-fold, P

Patients with non-alcoholic fatty liver disease are characterised by a decreased n-3/n-6 polyunsaturated fatty acid (PUFA) ratio in hepatic phospholipids. The metabolic consequences of n-3 PUFA depletion in the liver are poorly understood. We have reproduced a drastic drop in n-3 PUFA among hepatic phospholipids by feeding C57Bl/6J mice for 3 months with an n-3 PUFA depleted diet (DEF) versus a control diet (CT), which only differed in the PUFA content. DEF mice exhibited hepatic insulin resistance (assessed by euglycemic-hyperinsulinemic clamp) and steatosis that was associated with a decrease in fatty acid oxidation and occurred despite a higher capacity for triglyceride secretion. Microarray and qPCR analysis of the liver tissue revealed higher expression of all the enzymes involved in lipogenesis in DEF mice compared to CT mice, as well as increased expression and activation of sterol regulatory element binding protein-1c (SREBP-1c). Our data suggest that the activation of the liver X receptor pathway is involved in the overexpression of SREBP-1c, and this phenomenon cannot be attributed to insulin or to endoplasmic reticulum stress responses. In conclusion, n-3 PUFA depletion in liver phospholipids leads to activation of SREBP-1c and lipogenesis, which contributes to hepatic steatosis. © 2011 Pachikian et al., SCOPUS: ar.j, info:eu-repo/semantics/published

BACKGROUND: Anabolic resistance is the inability to increase protein synthesis in response to an increase in amino acids following a meal. One potential mediator of anabolic resistance is endoplasmic reticulum (ER) stress. The purpose of the present study was to test whether ER stress impairs the response to growth factors and leucine in muscle cells. METHODS: Muscle cells were incubated overnight with tunicamycin or thapsigargin to induce ER stress and the activation of the unfolded protein response, mTORC1 activity at baseline and following insulin and amino acids, as well as amino acid transport were determined. RESULTS: ER stress decreased basal phosphorylation of PKB and S6K1 in a dose-dependent manner. In spite of the decrease in basal PKB phosphorylation, insulin (10-50 nM) could still activate both PKB and S6K1. The leucine (2.5-5 mM)-induced phosphorylation of S6K1 on the other hand was repressed by low concentrations of both tunicamycin and thapsigargin. To determine the mechanism underlying this anabolic resistance, several inhibitors of mTORC1 activation were measured. Tunicamycin and thapsigargin did not change the phosphorylation or content of either AMPK or JNK, both increased TRB3 mRNA expression and thapsigargin increased REDD1 mRNA. Tunicamycin and thapsigargin both decreased the basal phosphorylation state of PRAS40. Neither tunicamycin nor thapsigargin prevented phosphorylation of PRAS40 by insulin. However, since PKB is not activated by amino acids, PRAS40 phosphorylation remained low following the addition of leucine. Blocking PKB using a specific inhibitor had the same effect on both PRAS40 and leucine-induced phosphorylation of S6K1. CONCLUSION: ER stress induces anabolic resistance in muscle cells through a PKB/PRAS40-induced blockade of mTORC1. ispartof: PLoS One vol:6 issue:6 ispartof: location:United States status: published

INTRODUCTION: Our goal was to determine whether in vivo administration of the proteasome inhibitor MG132 can prevent muscle atrophy caused by hindlimb unloading (HU). METHODS: Twenty-seven NMRI mice were assigned to a weight-bearing control, a 6-day HU, or a HU+MG132 (1 mg/kg/48 h) treatment group. RESULTS: Gastrocnemius wasting was significantly less in HU+MG132 mice (-6.7 ± 2.0%) compared with HU animals (-12.6 ± 1.1%, P = 0.011). HU was also associated with an increased expression of MuRF-1 (P = 0.006), MAFbx (P = 0.001), and USP28 (P = 0.027) mRNA, whereas Nedd4, E3α, USP19, and UBP45 mRNA did not change significantly. Increases in MuRF-1, MAFbx, and USP28 mRNA were largely repressed after MG132 administration. β5 proteasome activity tended to increase in HU (+16.7 ± 6.1%, P = 0.086). Neither β1 and β2 proteasome activities nor ubiquitin-conjugated proteins were changed by HU. CONCLUSIONS: Our results indicate that in vivo administration of MG132 partially prevents muscle atrophy associated with disuse and highlight an unexpected regulation of MG132 proteasome inhibitor on ubiquitin-ligases. ispartof: Muscle & Nerve vol:43 issue:5 pages:708-716 ispartof: location:United States status: published

This study investigated whether a 200-km run modulates signaling pathways implicated in cellular stress in skeletal muscle, with special attention paid to the endoplasmic reticulum (ER) stress and to the activation of the ubiquitin-proteasome pathway.Eight men ran 200 km (28 h 03 min ± 2 h 01 min). Two muscle biopsies were obtained from the vastus lateralis muscle 2 wk before and 3 h after the race. Mitogen-activated protein kinase, ubiquitin-proteasome pathway, ER stress, inflammation, and oxidative stress markers were assayed by Western blot analysis or by quantitative real-time polymerase chain reaction. Chymotrypsin-like activity of the proteasome was measured by a fluorimetric assay.Phosphorylation states of extracellular signal-related kinase 1/2 (+401% ± 173.8%, P = 0.027) and c-Jun N-terminal (+149% ± 61.9%, P = 0.023) increased after the race, whereas p38 phosphorylation remained unchanged. Increases in BiP (+235% ± 94.7%, P = 0.021) and in the messenger RNA level of total (+138% ± 31.2%, P = 0.002) and spliced X-box binding protein 1 (+241% ± 53.3%, P = 0.001) indicated the presence of ER stress. Transcripts of inflammatory markers interleukin-6 (+403% ± 96.1%, P = 0.002) and tumor necrosis factor-α (+233% ± 58.4%, P = 0.003) as well as oxidative stress markers metallothionein 1F (+519% ± 258.3%, P = 0.042), metallothionein 1H (+666% ± 157.5%, P = 0.002), and nicotinamide adenine dinucleotide phosphate-oxidase (NADPH oxidase) (+162% ± 60.5%, P = 0.016) were increased. The messenger RNA level of the ubiquitin ligases muscle-specific RING finger 1 (+583% ± 244.3%, P = 0.024) and muscle atrophy F-box (+249% ± 83.8%, P = 0.011) and the C2 proteasome subunit (+116% ± 40.6%, P = 0.012) also increased. Surprisingly, the amount of ubiquitin-conjugated proteins and the chymotrypsin-like activity of the proteasome were decreased by 20% ± 8.3% (P = 0.025) and 21% ± 4.4% (P = 0.001), respectively. The expression of ubiquitin-specific protease 28 deubiquitinase was increased (+81% ± 37.9%, P = 0.034).In the skeletal muscle, a 200-km run activates the expression of ubiquitin ligases muscle-specific RING finger 1 and muscle atrophy F-box as well as various cellular stresses, among which are ER stress, oxidative stress, and inflammation. Meanwhile, compensatory mechanisms seem also triggered: the unfolded protein response is up-regulated, and the chymotrypsin-like activity of the proteasome is repressed.

Magnesium (Mg) deficiency is a common nutritional disorder that is linked to an inflammatory state characterized by increased plasma acute phase protein and proinflammatory cytokine concentrations. Recent studies have shown that changes in the composition of gut microbiota composition participate in systemic inflammation. In this study, therefore, we assessed the potential role of gut microbiota in intestinal and systemic inflammation associated with Mg deficiency in mice. For this purpose, mice were fed a control or Mg-deficient diet (500 mg vs. 70 mg Mg/kg) for 4 or 21 d. Compared with the mice fed the control diet, mice fed the Mg-deficient diet for 4 d had a lower gut bifidobacteria content (-1.5 log), a 36-50% lower mRNA content of factors controlling gut barrier function in the ileum (zonula occludens-1, occludin, proglucagon), and a higher mRNA content (by approximately 2-fold) in the liver and/or intestine of tumor necrosis factor-alpha, interleukin-6, CCAAT/enhancer binding protein homologous protein, and activating transcription factor 4, reflecting inflammatory and cellular stress. In contrast, mice fed the Mg-deficient diet for 21 d had a higher cecal bifidobacteria content compared with the control group, a phenomenon accompanied by restoration of the intestinal barrier and the absence of inflammation. In conclusion, we show that Mg deficiency, independently of any other changes in nutrient intake, modulates the concentration of bifidobacteria in the gut, a phenomenon that may time-dependently affect inflammation and metabolic disorders in mice. ispartof: The Journal of Nutrition vol:140 issue:3 pages:509-514 ispartof: location:United States status: published

The present study aimed at comparing the responses of myogenic regulatory factors and signaling pathways involved in muscle protein synthesis after a resistance training session performed in either the fasted or fed state. According to a randomized crossover study design, six young male subjects participated in two experimental sessions separated by 3 weeks. In each session, they performed a standardized resistance training. After the sessions, they received during a 4-h recovery period 6 ml/kg b.w. h of a solution containing carbohydrates (50 g/l), protein hydrolysate (33 g/l), and leucine (16.6 g/l). On one occasion, the resistance exercise session was performed after the intake of a carbohydrate-rich breakfast (B), whereas in the other session they remained fasted (F). Needle biopsies from m. vastus lateralis were obtained before (Rest), and 1 h (+1h) and 4 h (+4h) after exercise. Myogenin, MRF4, and MyoD1 mRNA contents were determined by RT-PCR. Phosphorylation of PKB (protein kinase B), GSK3, p70(s6k) (p70 ribosomal S6 kinase), eIF2B, eEF2 (eukaryotic elongation factor 2), ERK1/2, and p38 was measured via western blotting. Compared with F, the pre-exercise phosphorylation states of PKB and p70(s6k) were higher in B, whereas those of eIF2B and eEF2 were lower. During recovery, the phosphorylation state of p70(s6k) was lower in B than in F (p = 0.02). There were no differences in basal mRNA contents between B and F. However, compared with F at +1h, MyoD1 and MRF4 mRNA contents were lower in B (p < 0.05). Our results indicate that prior fasting may stimulate the intramyocellular anabolic response to ingestion of a carbohydrate/protein/leucine mixture following a heavy resistance training session.

PURPOSE: To test whether endoplasmic reticulum (ER) stress impairs the response to growth factors and leucine in muscle cells. METHODS: C2C12 muscle cells were grown and fully differentiated into myotubes. After 4 days of differentiation, tunicamycin (TN) or thapsigargin (TG), known inducers of ER stress, were added for 17h before stimulation of the mTORC1 pathway with insulin or IGF-1 (10-50nM) for 15min or leucine (2.5-5mM) for 30min. Cells were harvested for mRNA or protein expression analyses or used for leucine uptake measurements. RESULTS: TN (0-5000ng/ml) and TG (0-2000nM) induced ER stress in a dose-dependent manner. At the end of the 17h-incubation, BiP and IRE1 expression and PERK and eIF2· phosphorylation were increased proportionally to TN and TG concentrations. At the same time, TN and TG repressed basal phosphorylation of PKB and S6K1. Growth factor-induction of PKB was repressed by low and high concentrations of TN but only by high concentrations of TG. Leucine-induction of S6K1 was repressed by low concentrations of both TN and TG. Leucine-induction of S6K1 was repressed at lower TG concentrations than growth factor-induction, indicating that TG inhibits leucine and growth factors activation of mTORC1 by different means. The activation/amount of several targets known to inhibit the mTORC1 pathway was determined. TN and TG did not change the phosphorylation state of AMPK or JNK. Both increased TRB3 mRNA expression and only TG increased REDD1 mRNA. TN and TG decreased the basal phosphorylation state of PRAS40. Interestingly, neither TN nor TG prevented the activation of PRAS40 by insulin. In contrast, PRAS40 phosphorylation did not increase with amino acids and TN and TG treatment further reduced its phosphorylation. Since non-phosphorylated PRAS40 inhibits mTORC1, this could impair downstream signaling by the mTORC1 complex. Finally the inhibition of leucine-induced phosphorylation of S6K1 could not be explained by a reduced entry of leucine into the cells as leucine uptake tended to increase with TN and TG. In summary, these data indicate that inhibition of PRAS40 by TN and TG could result in impaired activation of mTORC1 in response to leucine. CONCLUSIONS: TN and TG repressed the activation of mTORC1 by leucine and growth factors in muscle cells. As ER stress is triggered by high-fat feeding in skeletal muscle, we speculate that ER stress could be one mechanism impairing the response to load-induced hypertrophy in high-fat fed models.

Oxidative stress and endoplasmic reticulum stress (ER stress) are both involved in a number of diseases amongst which insulin resistance, Parkinson syndrome and atherosclerosis. Although, those stresses are often regarded in a separately way, it is known that they can be interrelated. Oxidative stress could lead to ER stress and vice versa as demonstrated by studies focused on the potential cross talk between the signaling pathways triggered by each of them in various cell types including mouse fibroblast cells, Chinese hamster ovary cells and mesencephalic cells. Skeletal muscle is a great provider of reactive oxygen species (ROS) and ER stress has been found to be present in skeletal muscle in certain conditions such as high-fat feeding and extreme endurance exercise. In agreement with those observations we have investigated the consequences of oxidative stress on ER stress in C2C12 myotubes. PURPOSE: To establish if oxidative stress, induced by hydrogen peroxide (H2O2), can lead to ER stress in C2C12 myotubes. METHODS: Myogenic C2C12 cells were grown until confluence and further differentiated for 4 days. Preliminary experiments were carried out to determine the optimal H2O2 dose and incubation period for inducing ER stress. A dose of 200μM and incubation periods of 4h and 17h were the most potent conditions for triggering ER stress and were used subsequently. Several downstream targets of the IRE1α and PERK pathways were analyzed at the protein level by western blot and at the mRNA level by qPCR. Three independent experiments were carried out. Unpaired Student t-test was used for the statistical analysis. RESULTS: After 4h H2O2 incubation, P-eIF2 α increased 7.7 +/- 1.3 fold (p0.05). At the mRNA level, bip and atf4 increased 1.9 +/- 0.2 and 2.4 +/- 0.4 fold, respectively (p

For over 300 years, scientists have understood that stimulation, in the form of an electrical impulse, is required for normal muscle function. More recently, the role of specific parameters of the electrical impulse (i.e., the pulse amplitude, pulse width, and work-to-rest ratio) has become better appreciated. However, most existing bioreactor systems do not permit sufficient control over these parameters. Therefore, the aim of the current study was to engineer an inexpensive muscle electrical stimulation bioreactor to apply physiologically relevant electrical stimulation patterns to tissue-engineered muscles and monolayers in culture. A low-powered microcontroller and a DC-DC converter were used to power a pulse circuit that converted a 4.5 V input to outputs of up to 50 V, with pulse widths from 0.05 to 4 ms, and frequencies up to 100 Hz (with certain operational limitations). When two-dimensional cultures were stimulated at high frequencies (100 Hz), this resulted in an increase in the rate of protein synthesis (at 12 h, control [CTL] = 5.0 + or - 0.16; 10 Hz = 5.0 + or - 0.07; and 100 Hz = 5.5 + or - 0.13 fmol/min/mg) showing that this was an anabolic signal. When three-dimensional engineered muscles were stimulated at 0.1 ms and one or two times rheobase, stimulation improved force production (CTL = 0.07 + or - 0.009; 1.25 V/mm = 0.10 + or - 0.011; 2.5 V/mm = 0.14146 + or - 0.012; and 5 V/mm = 0.03756 + or - 0.008 kN/mm(2)) and excitability (CTL = 0.53 + or - 0.022; 1.25 V/mm = 0.44 + or - 0.025; 2.5 V/mm = 0.41 + or - 0.012; and 5 V/mm = 0.60 + or - 0.021 V/mm), suggesting enhanced maturation. Together, these data show that the physiology and function of muscles can be improved in vitro using a bioreactor that allows the control of pulse amplitude, pulse width, pulse frequency, and work-to-rest ratio. ispartof: Tissue Engineering Part C, Methods vol:16 issue:4 pages:711-718 ispartof: location:United States status: published

PURPOSE:: Creatine (Cr) supplementation may improve muscle functional capacity in patients with neuromuscular diseases, disuse atrophy, or muscular dystrophies. Activation of myogenic satellite cells has been reported to be enhanced by Cr both in vitro and in vivo. Therefore, we hypothesized that Cr supplementation may improve the early steps of regeneration after muscle injury and may accelerate the recovery of both muscle mass and phenotype. METHODS:: Degeneration of left soleus muscle was induced by notexin injection in rats supplemented or not with Cr. The mass of regenerated muscles was compared with contralateral intact muscles at days 1, 3, 7, 14, 21, 28, 35, and 42 after injury. We also studied protein levels of the proliferator cell nuclear antigen (PCNA) as a marker of cell proliferation, expression of myogenic regulatory factors (MRF) as a marker of differentiation, and the myosin heavy chain (MHC) profile and activities of citrate synthase (CS) and lactate dehydrogenase (LDH) isozymes as markers of muscle phenotype maturation. RESULTS:: Cr supplementation accelerated the recovery of muscle Cr content during the regeneration phase. Although there were no other differences between Cr-treated and nontreated rats, we observed that 1) regenerated muscle mass remained lower than that in intact muscle mass 42 d after injury, 2) PCNA and MRF expression strongly increased in regenerated muscles, 3) the MHC profile of regenerated muscles was recovered 28 d after injury, and 4) CS activity was fully recovered from day 14, whereas the specific H isozyme of lactate dehydrogenase activity remained lower than that in intact muscles until 42 d. CONCLUSIONS:: In contrast with results from in vitro studies, Cr supplementation had no effects in vivo on the time course of recovery of rat skeletal muscle mass and phenotype after notexin-induced injury.

This study compared the effects of leucine and glutamine on the mTOR pathway, on protein synthesis and on muscle-specific gene expression in myogenic C(2)C(12) cells. Leucine increased the phosphorylation state of mTOR, on both Ser2448 and Ser2481, and its downstream effectors, p70(S6k), S6 and 4E-BP1. By contrast, glutamine decreased the phosphorylation state of mTOR on Ser2448, p70(S6k) and 4E-BP1, but did not modify the phosphorylation state of mTOR on Ser2481 and S6. Whilst the phosphorylation state of the mTOR pathway is usually related to protein synthesis, the incorporation of labelled methionine/cysteine was only transiently modified by leucine and was unaltered by glutamine. However, these two amino acids affected the mRNA levels of desmin, myogenin and myosin heavy chain in a time-dependant manner. In conclusion, leucine and glutamine have opposite effects on the mTOR pathway. Moreover, they induce modification of muscle-specific gene expression, unrelated to their effects on the mTOR/p70(S6k) pathway. ispartof: Amino Acids vol:35 issue:1 pages:147-55 ispartof: location:Austria status: published

To test the hypothesis that creatine supplementation would enhance the anabolic responses of muscle cell signaling and gene expression to exercise, we studied nine subjects who received either creatine or a placebo (maltodextrin) for 5 days in a double-blind fashion before undergoing muscle biopsies: at rest, immediately after exercise (10 x 10 repetitions of one-leg extension at 80% 1 repetition maximum), and 24 and 72 h later (all in the morning after fasting overnight). Creatine supplementation decreased the phosphorylation state of protein kinase B (PKB) on Thr308 at rest by 60% (P < 0.05) and that of eukaryotic initiation factor 4E-binding protein on Thr37/46 (4E-BP1) by 30% 24 h postexercise (P < 0.05). Creatine increased mRNA for collagen 1 (alpha(1)), glucose transporter-4 (GLUT-4), and myosin heavy chain I at rest by 250%, 45%, and 80%, respectively, and myosin heavy chain IIA (MHCIIA) mRNA immediately after exercise by 70% (all P < 0.05). Immediately after exercise, and independent of creatine, mRNA for muscle atrophy F-box (MAFbx), MHCIIA, peroxisome proliferator-activated receptor gamma coactivator-1alpha, and interleukin-6 were upregulated (60-350%; P < 0.05); the phosphorylation state of p38 both in the sarcoplasm and nucleus were increased (12- and 25-fold, respectively; both P < 0.05). Concurrently, the phosphorylation states of PKB (Thr308) and 4E-BP1 (Thr37/46) were decreased by 50% and 75%, respectively (P < 0.05). Twenty-four hours postexercise, MAFbx, myostatin, and GLUT-4 mRNA expression decreased below preexercise values (-35 to -50%; P < 0.05); calpain 1 mRNA increased 70% 72 h postexercise (P < 0.05) and at no other time. In conclusion, 5 days of creatine supplementation do not enhance anabolic signaling but increase the expression of certain targeted genes. ispartof: Journal of Applied Physiology vol:104 issue:2 pages:371-378 ispartof: location:United States status: published

SCOPUS: ar.j, info:eu-repo/semantics/published

The aim of the present study was to test if the consumption of creatine incorporated in food bars modifies creatine plasma kinetics, erythrocyte retention and loss in urine and in feces when compared with its consumption in the form of an aqueous solution (AS). Seventeen healthy young men ingested 2 g creatine either in the form of AS, or incorporated in a protein (PP)- or in a beta-glucan (BG)-rich food bar. Kinetics of plasma creatine was measured for 8-h duration and urinary excretion for 24 h. Then, the subjects received the same treatment thrice a day for 1 week at the end of which creatine contents were determined in erythrocytes and in feces (n = 4 for feces). The three crossover treatments were interspaced by a 40 +/- 1.2-day wash-out. Absorption of creatine was slowed down by 8-fold in the presence of BG (P < 0.001) and by 4-fold with PP (P < 0.001) whereas the velocity rate constant of elimination and the area under the curve were not modified. Urinary loss of creatine in the first 24 h following ingestion was 15 +/- 1.9% in AS and 14 +/- 2.2% in PP conditions (NS), whereas it was only 8 +/- 1.2% with BG (P = 0.004). Increase in creatine concentration in erythrocyte was similar in whatever form the creatine was ingested. Creatine seems to be totally absorbed since no creatine or creatinine was detectable in feces. No side effects were reported. In conclusion, ingestion of creatine combined with BG facilitates its retention by slowing down its absorption rate and reducing its urinary excretion. ispartof: European Journal of Applied Physiology vol:102 issue:2 pages:133-143 ispartof: location:Germany status: published

Objective : Muscle carnosine and its methylated form anserine are histidine-containing dipeptides. Both dipeptides have the ability to quench reactive carbonyl species and previous studies have shown that endogenous tissue levels are decreased in chronic diseases, such as diabetes. Design and Methods : Rodent study: Skeletal muscles of rats and mice were collected from 4 different diet-intervention studies, aiming to induce various degrees of glucose intolerance: 45% high-fat feeding (male rats), 60% high-fat feeding (male rats), cafeteria feeding (male rats), 70% high-fat feeding (female mice). Body weight, glucose-tolerance and muscle histidine-containing dipeptides were assessed. Human study: Muscle biopsies were taken from m. vastus lateralis in 35 males (9 lean, 8 obese, 9 prediabetic and 9 newly diagnosed type 2 diabetic patients) and muscle carnosine and gene expression of muscle fiber type markers were measured. Results : Diet interventions in rodents (cafeteria and 70% high-fat feeding) induced increases in body weight, glucose intolerance and levels of histidine-containing dipeptides in muscle. In humans, obese, prediabetic and diabetic men had increased muscle carnosine content compared to the lean (+21% (p>0.1), +30% (p

Needle biopsies are being extensively used in clinical trials addressing muscular adaptation to exercise and diet. Still, the potential artifacts due to biopsy sampling are often overlooked. Healthy volunteers (n = 9) underwent two biopsies through a single skin incision in a pretest. Two days later (posttest) another biopsy was taken 3 cm proximally and 3 cm distally to the pretest incision. Muscle oxygenation status (tissue oxygenation index [TOI]) was measured by near‐infrared spectroscopy. Biopsy samples were analyzed for 40 key markers (mRNA and protein contents) of myocellular O2 sensing, inflammation, cell proliferation, mitochondrial biogenesis, protein synthesis and breakdown, oxidative stress, and energy metabolism. In the pretest, all measurements were identical between proximal and distal biopsies. However, compared with the pretest, TOI in the posttest was reduced in the proximal (−10%, P < 0.05), but not in the distal area. Conversely, most inflammatory markers were upregulated at the distal (100–500%, P < 0.05), but not at the proximal site. Overall, 29 of the 40 markers measured, equally distributed over all pathways studied, were either up‐ or downregulated by 50–500% (P < 0.05). In addition, 19 markers yielded conflicting results between the proximal and distal measurements (P < 0.05). This study clearly documents that prior muscle biopsies can cause major disturbances in myocellular signaling pathways in needle biopsies specimens sampled 48 h later. In addition, different biopsy sites within identical experimental conditions yielded conflicting results., This study clearly demonstrates that skeletal muscle biopsying per se, at least by causing local tissue inflammation and/or topical deoxygenation, can substantially alter biochemical events happening in needle biopsy specimens sampled at a later day in the same muscle belly. It is crucial to take into account these potential artifacts whenever investigating the cellular mechanisms implicated in adaptation to exercise, recovery, or hypoxia.

A high turnover rate of intramyocellular lipids (IMCL) is proposed to reduce high-fat diet-induced accumulation of toxic lipid intermediates interfering with insulin action. It has been shown that an acute exercise bout in the fasted state stimulates IMCL breakdown compared with postprandial exercise. PURPOSE: To compare the effect of an exercise training program in the fasted state versus the postprandial state on insulin sensitivity and IMCL content during a period of hypercaloric fat-rich diet, and to explore the underlying cellular mechanisms. METHODS: Moderately active males (18-25y) consumed a hyper-caloric (normal diet +30% kcal/day) high-fat (50%) diet (HFD) during 6 weeks, alone (CON; n=7) or in conjunction with endurance training (1-1.5h @70% VO2max, 4d/week) in either the fasted state (F; n=10) or with carbohydrate intake before and during exercise (CHO; n=10). Needle biopsies were taken from m. vastus lateralis. Glucose tolerance was measured by a 2h oral glucose tolerance test. RESULTS: In CON, body weight on average increased by ∼3kg. This weight gain was largely, if not entirely accounted for by fat accretion as evidenced by the 15% increase in sum of skinfolds, reflecting elevated subcutaneous fat deposit. The HFD-induced increase in body weight and skinfolds was largely negated by training in F (P

Introduction Les acides gras polyinsatures (AGPI) n-3 jouent un role important sur le metabolisme lipidique de part leur capacite a stimuler la β-oxydation et a diminuer la lipogenese. Une augmentation du ratio n-6/n-3 est observee dans les phospholipides du foie de patientes atteintes de steatose hepatique non alcoolique. L’implication d’une diminution en AGPI n-3 dans l’accumulation de lipides au niveau du foie a ete peu etudie jusqu’a ce jour. Cette etude a pour but d’explorer le mecanisme implique dans l’accumulation des triglycerides dans le foie chez la souris carencee en AGPI n-3. Materiels et Methodes 24 souris mâles C57Bl6J ont ete nourries avec une diete controle (n-6/n-3 = 6,9) ou une diete carencee en AGPI n-3 (n-6/n-3 = 120) durant 3 mois. Resultats La carence en n-3 induit une accumulation de lipides dans le foie (triglyceride et cholesterol). L’incubation de tranches de foie en presence de 14 C-acetate et de 14 C-palmitate a revele une augmentation de la synthese et de l’esterification des acides gras dans le foie des souris carencees. La secretion hepatique de triglycerides, evaluee par inhibition de la lipoproteine lipase, est egalement augmentee chez les souris carencees. Les analyses microarrays demontrent, dans le foie des souris carencees, une augmentation de l’expression de toutes les enzymes impliquees dans la lipogenese et de SREBP-1c, un facteur de transcription cle dans le controle de leur expression. L’augmentation de l’expression de SREBP-1c ne peut pas etre expliquee par une augmentation de l’insulinemie, mais pourrait etre tributaire de l’activation de LXR (augmentation de l’expression d’ABCA1) et/ou du stress du reticulum (augmentation de la proteine d’IRE1α). Conclusion La carence en AGPI n-3 provoque une accumulation de lipides dans le foie suite a une stimulation de la lipogenese. SREBP-1c semble etre un facteur cle dans cette activation de la voie lipogenique et l’augmentation de son expression pourrait dependre de la stimulation de l’activite de LXR ou d’un stress du reticulum.

We hypothesize that the attenuated hypertrophic response in old mouse muscle is (1) partly due to a reduced capillarization and angiogenesis, which is (2) accompanied by a reduced oxidative capacity and fatigue resistance in old control and overloaded muscles, that (3) can be rescued by the antioxidant resveratrol. To investigate this, the hypertrophic response, capillarization, oxidative capacity, and fatigue resistance of m. plantaris were compared in 9- and 25-month-old non-treated and 25-month-old resveratrol-treated mice. Overload increased the local capillary-to-fiber ratio less in old (15 %) than in adult (59 %) muscle (P