The endocannabinoid system links gut microbiota to adipogenesis

We investigated several models of gut microbiota modulation: selective (prebiotics, probiotics, high-fat), drastic (antibiotics, germ-free mice) and mice bearing specific mutations of a key gene involved in the toll-like receptors (TLR) bacteria-host interaction (Myd88−/−). Here we report that gut microbiota modulates the intestinal endocannabinoid (eCB) system-tone, which in turn regulates gut permeability and plasma lipopolysaccharide (LPS) levels.The activation of the intestinal endocannabinoid system increases gut permeability which in turn enhances plasma LPS levels and inflammation in physiological and pathological conditions such as obesity and type 2 diabetes.The investigation of adipocyte differentiation and lipogenesis (both markers of adipogenesis) indicate that gut microbiota controls adipose tissue physiology through LPS-eCB system regulatory loops and may play a critical role in the adipose tissue plasticity during obesity.In vivo, ex vivo and in vitro studies indicate that LPS acts as a master switch on adipose tissue metabolism, by blocking the cannabinoid-driven adipogenesis

PL-012 Effect of hypoxic resistance training on the regulation of muscle mass and phenotype

Objective Hypoxia is a state of lowered oxygen tension in tissue that can be created by environmental or pathological conditions. Whatever the origin of hypoxia, different tissues will adapt acutely and/or chronically to deal with this reduction in oxygen availability. Hypoxia has recently emerged as a particularly efficient stimulus to stimulate muscle cell proliferation and accretion of muscle mass and hypoxic resistance training has become popular amongst athletes as it is thought to favor muscle accretion. However, the molecular mechanisms are largely unknown.  Methods To determine those molecular mechanisms, 19 volunteers participated to 12 sessions of resistance training spread over 4 weeks whether in normoxia (n=9) or in hypoxia (n=10, FiO213.5% corresponding to 3500m altitude). Each session consisted in 6 sets of 10 repetitions of a one-leg extension exercise at 80% of one repetition maximum (1-RM). Blood and muscle samples in each leg were taken before and after the 4-week training period. Fiber types were determined by immunohistochemistry based on myosin heavy chain isotypes. Blood saturation (SpO2, pulsoximetry) and tissue saturation index (TSI, near-infrared spectroscopy) were monitored during the exercise sessions.  Results Muscle thickness determined by ultrasound was increased by 7% in normoxia only (p=0.04). The 1-RM was increased in both groups but the increase was higher in hypoxia (+34%) than in normoxia (+24%) (p=0.02). In average, SpO2stayed around 98-99% in normoxia and around 93-94% in hypoxia during each set of contractions. No difference in TSI between normoxia and hypoxia was measured, which averaged 60% before starting muscle contractions and 40% during muscle contractions. A trend towards a shift in fiber type from type I to type IIa was observed in normoxia (p<0.09) but not in hypoxia. Fiber area was not modified by any condition.  Conclusions In summary, 4 weeks of hypoxic resistance training induced a larger increase in 1-RM compared to normoxic resistance training, independently of muscle hypertrophy or any change in fiber type. Further investigation should determine whether metabolic or molecular changes may explain this potentiation of maximal muscle force by hypoxia

Environmental hypoxia favors myoblast differentiation and fast phenotype but blunts activation of protein synthesis after resistance exercise in human skeletal muscle

We hypothesized that a single session of resistance exercise performed in moderate hypoxic (FiO2: 14%) environmental conditions would potentiate the anabolic response during the recovery period spent in normoxia. Twenty subjects performed a 1-leg knee extension session in normoxic or hypoxic conditions. Muscle biopsies were taken 15 min and 4 h after exercise in the vastus lateralis of the exercised and the nonexercised legs. Blood and saliva samples were taken at regular intervals before, during, and after the exercise session. The muscle fractional-protein synthetic rate was determined by deuterium incorporation into proteins, and the protein-degradation rate was determined by methylhistidine release from skeletalmuscle.Wefoundthat:1)hypoxiablunted the activation of protein synthesis after resistance exercise; 2) hypoxia down-regulated the transcriptional program of autophagy; 3) hypoxia regulated the expression of genes involved in glucose metabolism at rest and the genes involved in myoblast differentiation and fusion and in muscle contraction machinery after exercise; and 4) the hypoxia-inducible factor-1alpha pathway was not activated at the time points studied. Contrary to our hypothesis, environmental hypoxia did not potentiate the short-term anabolic response after resistance exercise, but it initiated transcriptional regulations that could potentially translate into satellite cell incorporation and higher force production in the long term

Higher activation of autophagy in skeletal muscle of mice during endurance exercise in the fasted state.

Activation of autophagy in skeletal muscle has been reported in response to endurance exercise and food deprivation independently. The purpose of this study was to evaluate whether autophagy was more activated when both stimuli were combined, namely when endurance exercise was performed in a fasted rather than a fed state. Mice performed a low-intensity running exercise (10 m/min for 90min) in both dietary states after which the gastrocnemius muscles were removed. LC3b-II, a marker of autophagosome presence, increased in both conditions, but the increase was higher in the fasted state. Other protein markers of autophagy, like Gabarapl1-II and Atg12 conjugated form as well as mRNA of Lc3b, Gabarapl1, and p62/Sqstm1 were increased only when exercise was performed in a fasted state. The larger activation of autophagy by exercise in a fasted state was associated with a larger decrease in plasma insulin and phosphorylation of Akt(Ser473), Akt(Thr308), FoxO3a(Thr32), and ULK1(Ser757). AMPKα(Thr172), ULK1(Ser317), and ULK1(Ser555) remained unchanged in both conditions, whereas p38(Thr180/Tyr182) increased during exercise to a similar extent in the fasted and fed conditions. The marker of mitochondrial fission DRP1(Ser616) was increased by exercise independently of the nutritional status. Changes in mitophagy markers BNIP3 and Parkin suggest that mitophagy was increased during exercise in the fasted state. In conclusion, our results highlight a major implication of the insulin-Akt-mTOR pathway and its downstream targets FoxO3a and ULK1 in the larger activation of autophagy observed when exercise is performed in a fasted state compared with a fed state

Regular Endurance Exercise Promotes Fission, Mitophagy, and Oxidative Phosphorylation in Human Skeletal Muscle Independently of Age.

This study investigated whether regular endurance exercise maintains basal mitophagy and mitochondrial function during aging. Mitochondrial proteins and total mRNA were isolated from vastus lateralis biopsies ( = 33) of young sedentary (YS), old sedentary (OS), young active (YA), and old active (OA) men. Markers for mitophagy, fission, fusion, mitogenesis, and mitochondrial metabolism were assessed using qRT-PCR, Western blot, and immunofluorescence staining. Independently of age, fission protein Fis1 was higher in active vs. sedentary subjects (+80%; < 0.05). Mitophagy protein PARKIN was more elevated in OA than in OS (+145%; = 0.0026). mRNA expression of Beclin1 and Gabarap, involved in autophagosomes synthesis, were lower in OS compared to YS and OA ( < 0.05). Fusion and oxidative phosphorylation proteins were globally more elevated in the active groups ( < 0.05), while COx activity was only higher in OA than in OS ( = 0.032). Transcriptional regulation of mitogenesis did not vary with age or exercise. In conclusion, physically active lifestyle seems to participate in the maintenance of lifelong mitochondrial quality control by increasing fission and mitophagy

Effects of Sprint Interval Training at Different Altitudes on Cycling Performance at Sea-Level

Background: Benefits of sprint interval training performed in hypoxia (SIH) compared to normoxia (SIN) have been assessed by studies mostly conducted around 3000 m of simulated altitude. The present study aims to determine whether SIH at an altitude as high as 4000 m can elicit greater adaptations than the same training at 2000 m, 3000 m or sea-level. Methods: Thirty well-trained endurance male athletes (18-35 years old) participated in a six-week repeated sprint interval training program (30 s all-out sprint, 4 min 30 s recovery; 4-9 repetitions, 2 sessions/week) at sea-level (SL, n = 8), 2000 m (FiO2 16.7%, n = 8), 3000 m (FiO2 14.5%, n = 7) or 4000 m (FiO2 13.0%, n = 7). Aerobic and anaerobic exercise components were evaluated by an incremental exercise test, a 600 kJ time trial and a Wingate test before and after the training program. Results: After training, peak power output (PPO) during the incremental exercise test increased (~6%) without differences between groups. The lactate threshold assessed by Dmax increased at 2000 m (+14 ± 12 W) and 4000 m (+12 ± 11 W) but did not change at SL and 3000 m. Mean power during the Wingate test increased at SL, 2000 m and 4000 m, although peak power increased only at 4000 m (+38 ± 38 W). Conclusions: The present study indicates that SIH using 30 s sprints is as efficient as SIN for improving aerobic and anaerobic qualities. Additional benefits such as lactate-related adaptations were found only in SIH and Wingate peak power only increased at 4000 m. This finding is of particular interest for disciplines requiring high power output, such as in very explosive sports

Toll-Like Receptor 4 Knockout Mice Are Protected against Endoplasmic Reticulum Stress Induced by a High-Fat Diet.

The purpose of this study was to investigate whether toll-like receptor 4 (TLR4) is implicated in the development of endoplasmic reticulum stress (ER stress) observed after a high-fat diet (HFD) in liver, skeletal muscle and adipose tissue. TLR4(-/-) and C57BL/6J wild-type mice (WT) were fed with chow or HFD (45% calories from fat) during 18 weeks. An oral glucose tolerance-test was performed. The animals were sacrificed in a fasted state and the tissues were removed. TLR4 deletion protected from body weight gain and glucose intolerance induced by HFD whereas energy intake was higher in transgenic mice suggesting larger energy expenditure. HFD induced an ER stress in skeletal muscle, liver and adipose tissue of WT mice as assessed by BiP, CHOP, spliced and unspliced XBP1 and phospho-eIF2α. TLR4(-/-) mice were protected against HFD-induced ER stress. Then, we investigated the main signaling downstream of TLR4 namely the NF-κB pathway, expecting to identify the mechanism by which TLR4 is able to activate ER stress. The mRNA levels of cytokines regulated by NF-κB namely TNFα, IL-1β and IL-6, were not changed after HFD and phospho-IκB-α (ser 32) was not changed. Our results indicate that TLR4 is essential for the development of ER stress related to HFD. Nevertheless, the NFκ-B pathway does not seem to be directly implicated. The reduced fat storage in TLR4(-/-) mice could explain the absence of an ER stress after HFD

Pomegranate and green tea extracts protect against ER stress induced by a high-fat diet in skeletal muscle of mice.

We tested the hypothesis that polyphenol-rich extracts can reduce endoplasmic reticulum (ER) stress induced by a high-fat diet (HFD) in skeletal muscle of mice