Muscle Fatty Acid Oxidation Research Articles

Investigating ZMP Accumulation in C2C12 Myotubes with AICAR and Prodrug-39

5’Adenosine monophosphate (AMP) is a signaling molecule and a known regulator of enzymes in glycolysis, glycogen breakdown, and may play a role in regulating protein synthesis. It is also a known activator of AMP-dependent protein kinase (AMPK), a key energy sensor within the cell. Activation of AMPK has been linked to the regulation of mitochondrial biogenesis, upregulation of BCAA catabolism, and increased skeletal muscle glucose uptake and fatty acid oxidation. AMPK can be activated pharmacologically with 5-Aminoimidazole-4-carboxamide ribonucleoside (AICAR), an adenosine analogue. AICAR enters the cell through adenosine transporters and undergoes phosphorylation by Adenosine Kinase forming AICAR monophosphate (ZMP). Unfortunately, the poor bioavailability of AICAR limits its use pharmacologically. Prodrug-39 (P39) is comprised of ZMP with a 4-nitrophenol group to allow membrane permeability and subsequent ZMP formation without the need of membrane transporters. We investigated the ZMP accumulation in C2C12 myotubes following exposure to different concentrations of P39 or AICAR. Based on preliminary data, we hypothesized that AICAR treatments would be more effective at producing ZMP over short periods of time and when treatment concentrations were high, but that P39 treatments would result in greater and more consistent ZMP accumulation over longer periods of time when treatment concentrations were low. We first measured the absorbance of 4-nitrophenol (4NP) within the cell media to determine phosphodiesterase cleavage of P39. Nitrophenol absorbance was 3.9 times higher at 1000 μM compared to 100 μM treatment of P39 after 15 minute incubation, and 13.3 times higher at 11 hours. The nitrophenol absorbance per hour increased linearly between 250 μM to 1000 μM of P39. Adenine nucleotide content and ZMP accumulation in treated cells was determined following organic protein extraction and UV-Vis ultra-performance liquid chromatography (UPLC). After 13 hours of treatment with 500 μM AICAR or P39, ZMP accumulation was 14 fold higher following AICAR treatment compared to P39 (59.7±5.0 vs 4.2±0.1 μmol/g protein). Interestingly, ZMP accumulation was not different between 100 μM treatments (5.6±1.3 vs. 5.7±1.5 μmol/g protein). This suggests that phosphodiesterase cleavage of P39 is likely the limiting factor for ZMP accumulation. Interestingly, while ZMP accumulation following 13 hour incubation with 100 μM AICAR or P39 was not different, ATP content was approximately 50% higher in P39 treated cells (33.7±5.8 vs. 51.0±5.8 μmol/g protein). While the measurement of ZMP accumulation at very low treatment concentrations (10 μM) is very diffcult, P39 exposure for 1 to 1.5 hours resulted in a 3 to 8 fold increase in ZMP content compared to AICAR. This data illustrates the potential for P39 as a drug capable of accumulating greater ZMP than AICAR at very low treatment concentrations, however the capacity of phosphodiesterase activity may limit the application of this drug. This work was funded by a grant from Skylark Biosciences. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Effects of dietary lipid levels on fiber quality, lipidomic profiles, antioxidant and inflammation responses in muscle of yellow catfish Pelteobagrus fulvidraco

The present study explore the impacts of dietary lipid levels on fiber quality, lipidomic profiles, antioxidant and inflammation responses in muscle of yellow catfish. For this purpose, yellow catfish were fed with low-fat (LF), medium-fat (MF) and high-fat (HF) three different lipid levels diets for 8 weeks, and at the end of the feeding trial, muscle tissues were taken for analysis. We found that HF diet increased lipid deposition, lipogenesis and decreased fatty acid oxidation in muscle of yellow catfish. HF diet induced oxidative stress and inflammatory response in muscle therefore suppressed the proliferation and differentiation of myofibril. Lipidomic analysis detected 229 differentially abundant lipids (DALs) among three groups, and PCs, TGs/DGs and PEs as the predominant lipid species. Moreover, HF diet significantly decreased the abundance of SHP, LPE, PC, PE, PI, PG and PS species. Overall, these findings revealed that dietary lipid levels influence lipid metabolism and muscle quality by affecting lipid profile and oxidative stress status in muscle of yellow catfish, which provided novel insight into the lipid nutrition of freshwater fish.

Stimulation of fat oxidation in rat muscle by unacylated ghrelin persists for 2-3 hours, but is independent of fatty acid transporter translocation.

While a definitive mechanism-of-action remains to be identified, recent findings indicate that ghrelin, particularly the unacylated form (UnAG), stimulates skeletal muscle fatty acid oxidation. The biological importance of UnAG-mediated increases in fat oxidation remains unclear, as UnAG peaks in the circulation before mealtimes, and decreases rapidly during the postprandial situation before increases in postabsorptive circulating lipids. Therefore, we aimed to determine if the UnAG-mediated stimulation of fat oxidation would persist long enough to affect the oxidation of meal-derived fatty acids, and if UnAG stimulated the translocation of fatty acid transporters to the sarcolemma as a mechanism-of-action. In isolated soleus muscle strips from male rats, short-term pre-treatment with UnAG elicited a persisting stimulus on fatty acid oxidation 2 h after the removal of UnAG. UnAG also caused an immediate phosphorylation of AMPK, but not an increase in plasma membrane FAT/CD36 or FABPpm. There was also no increase in AMPK signaling or increased FAT/CD36 or FABPpm content at the plasma membrane at 2 h which might explain the sustained increase in fatty acid oxidation. These findings confirm UnAG as a stimulator of fatty acid oxidation and provide evidence that UnAG may influence the handling of postprandial lipids. The underlying mechanisms are not known.

High-fat diet during pregnancy promotes fetal skeletal muscle fatty acid oxidation and insulin resistance in an ovine model.

Maternal diet during pregnancy is associated with offspring metabolic risk trajectory in humans and animal models, but the prenatal origins of these effects are less clear. We examined the effects of a high-fat diet (HFD) during pregnancy on fetal skeletal muscle metabolism and metabolic risk parameters using an ovine model. White-faced ewes were fed a standardized diet containing 5% fat wt/wt (CON), or the same diet supplemented with 6% rumen-protected fats (11% total fat wt/wt; HFD) beginning 2 wk before mating until midgestation (GD75). Maternal HFD increased maternal weight gain, fetal body weight, and low-density lipoprotein levels in the uterine and umbilical circulation but had no significant effects on circulating glucose, triglycerides, or placental fatty acid transporters. Fatty acid (palmitoylcarnitine) oxidation capacity of permeabilized hindlimb muscle fibers was >50% higher in fetuses from HFD pregnancies, whereas pyruvate and maximal (mixed substrate) oxidation capacities were similar to CON. This corresponded to greater triacylglycerol content and protein expression of fatty acid transport and oxidation enzymes in fetal muscle but no significant effect on respiratory chain complexes or pyruvate dehydrogenase expression. However, serine-308 phosphorylation of insulin receptor substrate-1 was greater in fetal muscle from HFD pregnancies along with c-jun-NH2 terminal kinase activation, consistent with prenatal inhibition of skeletal muscle insulin signaling. These results indicate that maternal high-fat feeding shifts fetal skeletal muscle metabolism toward a greater capacity for fatty acid over glucose utilization and favors prenatal development of insulin resistance, which may predispose offspring to metabolic syndrome later in life.NEW & NOTEWORTHY Maternal diet during pregnancy is associated with offspring metabolic risk trajectory in humans and animal models, but the prenatal origins of these effects are less clear. This study examined the effects of a high-fat diet during pregnancy on metabolic risk parameters using a new sheep model. Results align with findings previously reported in nonhuman primates, demonstrating changes in fetal skeletal muscle metabolism that may predispose offspring to metabolic syndrome later in life.

Synthesis of 2′,4′,6′-trimethoxy-3′,5′-dimethylchalcone derivatives and their anti-diabetic and anti-atherosclerosis effects

AMP-activated protein kinase (AMPK) is an important regulator of energy metabolism and is considered a promising target for the treatment of metabolic diseases including diabetes. However, no AMPK agonist is used clinically at present. Here, we synthesized various derivatives of 2′,4′,6′-trimethoxy-3′,5′-dimethylchalcone and tested whether they activated AMPK and improved glucose tolerance in an obese mouse model. The derivatives activated AMPK at a much lower concentration than 5-aminoimidazole-4-carboxamide ribonucleotide, a well-known AMPK agonist, in C2C12 myotubes. Among them, compounds 2c and 2i showed predominant AMPK activation effects and increased the fatty acid oxidation (FAO) rate more than 2-fold in C2C12 myotubes. When compound 2i was administered to high fat diet-induced obese mice for two weeks, the glucose tolerance was improved and skeletal muscle FAO rate increased (1.6-fold). In addition, several derivatives, including compound 2i, suppressed the migration of vascular smooth muscle cells treated with platelet-derived growth factor, and inhibited tumor necrosis factor-α-induced adhesion between monocytes and endothelial cells. These results suggest these derivatives have anti-atherosclerosis effects. Taken together, the derivatives, especially compound 2i, might be therapeutic agents against diabetes and atherosclerosis.

Skeletal muscle proteins involved in fatty acid transport influence fatty acid oxidation rates observed during exercise.

Several proteins are implicated in transmembrane fatty acid transport. The purpose of this study was to quantify the variation in fatty acid oxidation rates during exercise explained by skeletal muscle proteins involved in fatty acid transport. Seventeen endurance-trained males underwent a (i) fasted, incremental cycling test to estimate peak whole-body fatty acid oxidation rate (PFO), (ii) resting vastus lateralis microbiopsy, and (iii) 2 h of fed-state, moderate-intensity cycling to estimate whole-body fatty acid oxidation during fed-state exercise (FO). Bivariate correlations and stepwise linear regression models of PFO and FO during 0-30 min (early FO) and 90-120 min(late FO) of continuous cycling were constructed using muscle data. To assess the causal role of transmembrane fatty acid transport in fatty acid oxidation rates during exercise, we measured fatty acid oxidation during in vivo exercise and ex vivo contractions in wild-type and CD36 knock-out mice. We observed a novel, positive association between vastus lateralis FATP1 and PFO and replicated work reporting a positive association between FABPpm and PFO. The stepwise linear regression model of PFO retained CD36, FATP1, FATP4, and FABPpm, explaining ~87% of the variation. Models of early and late FO explained ~61 and ~65% of the variation, respectively. FATP1 and FATP4 emerged as contributors to models of PFO and FO. Mice lacking CD36 had impaired whole-body and muscle fatty acid oxidation during exercise and muscle contractions, respectively. These data suggest that substantial variation in fatty acid oxidation rates during exercise can be explained by skeletal muscle proteins involved in fatty acid transport.

Muscle Injuries Induce a Prostacyclin-PPARγ/PGC1a-FAO Spike That Boosts Regeneration.

It is well-known that muscle regeneration declines with aging, and aged muscles undergo degenerative atrophy or sarcopenia. While exercise and acute injury are both known to induce muscle regeneration, the molecular signals that help trigger muscle regeneration have remained unclear. Here, mass spectrometry imaging (MSI) is used to show that injured muscles induce a specific subset of prostanoids during regeneration, including PGG1, PGD2, and the prostacyclin PGI2. The spike in prostacyclin promotes skeletal muscle regeneration via myoblasts, and declines with aging. Mechanistically, the prostacyclin spike promotes a spike in PPARγ/PGC1a signaling, which induces a spike in fatty acid oxidation (FAO) to control myogenesis. LC-MS/MS and MSI further confirm that an early FAO spike is associated with normal regeneration, but muscle FAO became dysregulated during aging. Functional experiments demonstrate that the prostacyclin-PPARγ/PGC1a-FAO spike is necessary and sufficient to promote both young and aged muscle regeneration, and that prostacyclin can synergize with PPARγ/PGC1a-FAO signaling to restore aged muscles' regeneration and physical function. Given that the post-injury prostacyclin-PPARγ-FAO spike can be modulated pharmacologically and via post-exercise nutrition, this work has implications for how prostacyclin-PPARγ-FAO might be fine-tuned to promote regeneration and treat muscle diseases of aging.

The Effect of Physical Activity on the Secretory Function of Muscles and Bone Tissue in Humans

The purpose of the study was to summarize new literature data on the effect of physical activity on the secretory function of skeletal muscles and their relationship with organs. Materials and methods. Analytical methods were used in the study. The search for scientific information was carried out in databases of search systems. Results. Muscles secrete proteins called myokines, which are involved in a variety of processes by interacting with tissues. Current research has shown that exercise, by stimulating the skeletal muscle system in vivo, leads to the release of myokines and causes several effects that explain the positive effect of exercise in the treatment of several diseases of the musculoskeletal system. Skeletal muscles synthesize and secrete a wide range of myokines that contribute to various functions in organs, including the brain. Recent publications have focused much attention on one such myokine, the recently discovered protein irisin, which is secreted into the bloodstream from skeletal muscle during exercise from the membrane-bound precursor fibronectin type III. Irisin promotes metabolic processes such as glucose homeostasis and the darkening of white adipose tissue. Irisin also crosses the blood-brain barrier and initiates a neuroprotective genetic program in the hippocampus, which culminates in increased expression of brain-derived neurotrophic factors. Most studies report that irisin concentration is closely related to health status. For example, irisin levels are significantly lower in patients with obesity, osteoporosis/fracture, muscular atrophy, Alzheimer's disease, and cardiovascular disease. Over the past decade, several myokines have been discovered, expanding our understanding of how muscles interact with other organs. In recent publications, a lot of attention is paid to the recently discovered protein irisin, which contributes to metabolic processes. A series of studies on irisin have provided new insights into the mechanisms of exercise to improve bone density, counteract cartilage degeneration, and maintain overall joint environmental homeostasis. These studies further contribute to the understanding of the role of exercise in the fight against osteoarthritis and may provide important assistance in advancing the prevention and treatment of this common disease. Research on the exercise-induced muscle factor irisin will help to better understand and explain the beneficial effects of exercise on maintaining physical health, especially in the fight against aging and age-related degenerative changes. Conclusion. Thus, exercise-induced stimulation of bioactive cytokines increases muscle anabolism, bone formation, mitochondrial biogenesis, glucose utilization, and fatty acid oxidation, and reduces chronic inflammation

Targeting activin receptor-like kinase 7 ameliorates adiposity and associated metabolic disorders.

Activin receptor-like kinase 7 (ALK7) is a type I receptor in the TGF-β superfamily preferentially expressed in adipose tissue and associated with lipid metabolism. Inactivation of ALK7 signaling in mice results in increased lipolysis and resistance to both genetic and diet-induced obesity. Human genetic studies have recently revealed an association between ALK7 variants and both reduced waist to hip ratios and resistance to development of diabetes. In the present study, treatment with a neutralizing mAb against ALK7 caused a substantial loss of adipose mass and improved glucose intolerance and insulin resistance in both genetic and diet-induced mouse obesity models. The enhanced lipolysis increased fatty acid supply from adipocytes to promote fatty acid oxidation in muscle and oxygen consumption at the whole-body level. The treatment temporarily increased hepatic triglyceride levels, which resolved with long-term Ab treatment. Blocking of ALK7 signals also decreased production of its ligand, growth differentiation factor 3, by downregulating S100A8/A9 release from adipocytes and, subsequently, IL-1β release from adipose tissue macrophages. These findings support the feasibility of potential therapeutics targeting ALK7 as a treatment for obesity and diabetes.

Data Mining Evidences Variabilities in Glucose and Lipid Metabolism among Fish Strains: A Case Study on Three Genotypes of Gibel Carp Fed by Different Carbohydrate Sources.

An 8-week feeding trial was conducted to evaluate the application of common carbohydrate sources, cornstarch (CS), wheat starch (WS), and wheat flour (WF), to different gibel carp genotypes, Dongting, CASIII, and CASV. The results of the growth and physical responses were analysed by data visualization and unsupervised machine learning. As revealed by a self-organizing map (SOM) and the cluster of growth and biochemical indicators, CASV had superior growth and feed utilization and better regulation of postprandial glucose, followed by CASIII, while Dongting showed a high level of plasma glucose with poor growth performance. CS, WS, and WF were differently utilized by the gibel carp, and WF was associated with greater zootechnical performance based on higher specific growth rate (SGR), feed efficiency (FE), protein retention efficiency (PRE), and lipid retention efficiency (LRE), induced hepatic lipogenesis, increased liver lipids, and enhanced muscle glycogen. Spearman's correlation analysis of the physiological responses indicated that plasma glucose had a significantly negative correlation with growth, feed utilization, glycogen storage, and plasma cholesterol level, and it was positively related to liver fat content in gibel carp. Transcriptional variabilities were observed: CASIII showed increased expression of pklr, which is involved in hepatic glycolysis, and pck and g6p, which are involved in gluconeogenesis. Interestingly, Dongting showed upregulation of genes involved in glycolysis and fatty acid oxidation in muscle. Furthermore, there were numerous interactions between carbohydrate sources and strains for growth, metabolites, and transcriptional control, confirming the existence of genetic polymorphisms in carbohydrate use in gibel carp. Globally, CASV showed relatively better growth and carbohydrate utilization, and wheat flour seemed to be more efficiently utilized by gibel carp.

Melatonin supplementation promotes muscle fiber hypertrophy and regulates lipid metabolism of skeletal muscle in weaned piglets.

Melatonin has been reported to play crucial roles in regulating meat quality, improving reproductive properties, and maintaining intestinal health in animal production, but whether it regulates skeletal muscle development in weaned piglet is rarely studied. This study was conducted to investigate the effects of melatonin on growth performance, skeletal muscle development, and lipid metabolism in animals by intragastric administration of melatonin solution. Twelve 28-d-old DLY (Duroc × Landrace × Yorkshire) weaned piglets with similar body weight were randomly divided into two groups: control group and melatonin group. The results showed that melatonin supplementation for 23 d had no effect on growth performance, but significantly reduced serum glucose content (P < 0.05). Remarkably, melatonin increased longissimus dorsi muscle (LDM) weight, eye muscle area and decreased the liver weight in weaned piglets (P < 0.05). In addition, the cross-sectional area of muscle fibers was increased (P < 0.05), while triglyceride levels were decreased in LDM and psoas major muscle by melatonin treatment (P < 0.05). Transcriptome sequencing showed melatonin induced the expression of genes related to skeletal muscle hypertrophy and fatty acid oxidation. Enrichment analysis indicated that melatonin regulated cholesterol metabolism, protein digestion and absorption, and mitophagy signaling pathways in muscle. Gene set enrichment analysis also confirmed the effects of melatonin on skeletal muscle development and mitochondrial structure and function. Moreover, quantitative real-time polymerase chain reaction analysis revealed that melatonin supplementation elevated the gene expression of cell differentiation and muscle fiber development, including paired box 7 (PAX7), myogenin (MYOG), myosin heavy chain (MYHC) IIA and MYHC IIB (P < 0.05), which was accompanied by increased insulin-like growth factor 1 (IGF-1) and insulin-like growth factor binding protein 5 (IGFBP5) expression in LDM (P < 0.05). Additionally, melatonin regulated lipid metabolism and activated mitochondrial function in muscle by increasing the mRNA abundance of cytochrome c oxidase subunit 6A (COX6A), COX5B, and carnitine palmitoyltransferase 2 (CPT2) and decreasing the mRNA expression of peroxisome proliferator-activated receptor gamma (PPARG), acetyl-CoA carboxylase (ACC) and fatty acid-binding protein 4 (FABP4) (P < 0.05). Together, our results suggest that melatonin could promote skeletal muscle growth and muscle fiber hypertrophy, improve mitochondrial function and decrease fat deposition in muscle.

Evidence for a neuromuscular circuit involving hypothalamic interleukin-6 in the control of skeletal muscle metabolism.

Hypothalamic interleukin-6 (IL6) exerts a broad metabolic control. Here, we demonstrated that IL6 activates the ERK1/2 pathway in the ventromedial hypothalamus (VMH), stimulating AMPK/ACC signaling and fatty acid oxidation in mouse skeletal muscle. Bioinformatics analysis revealed that the hypothalamic IL6/ERK1/2 axis is closely associated with fatty acid oxidation– and mitochondrial-related genes in the skeletal muscle of isogenic BXD mouse strains and humans. We showed that the hypothalamic IL6/ERK1/2 pathway requires the α2-adrenergic pathway to modify fatty acid skeletal muscle metabolism. To address the physiological relevance of these findings, we demonstrated that this neuromuscular circuit is required to underpin AMPK/ACC signaling activation and fatty acid oxidation after exercise. Last, the selective down-regulation of IL6 receptor in VMH abolished the effects of exercise to sustain AMPK and ACC phosphorylation and fatty acid oxidation in the muscle after exercise. Together, these data demonstrated that the IL6/ERK axis in VMH controls fatty acid metabolism in the skeletal muscle.

Short-term mercury exposure disrupts muscular and hepatic lipid metabolism in a migrant songbird

Methylmercury (MeHg) is a global pollutant that can cause metabolic disruptions in animals and thereby potentially compromise the energetic capacity of birds for long-distance migration, but its effects on avian lipid metabolism pathways that support endurance flight and stopover refueling have never been studied. We tested the effects of short-term (14-d), environmentally relevant (0.5 ppm) dietary MeHg exposure on lipid metabolism markers in the pectoralis and livers of yellow-rumped warblers (Setophaga coronata) that were found in a previous study to have poorer flight endurance in a wind tunnel than untreated conspecifics. Compared to controls, MeHg-exposed birds displayed lower muscle aerobic and fatty acid oxidation capacity, but similar muscle glycolytic capacity, fatty acid transporter expression, and PPAR expression. Livers of exposed birds indicated elevated energy costs, lower fatty acid uptake capacity, and lower PPAR-γ expression. The lower muscle oxidative enzyme capacity of exposed birds likely contributed to their weaker endurance in the prior study, while the metabolic changes observed in the liver have potential to inhibit lipogenesis and stopover refueling. Our findings provide concerning evidence that fatty acid catabolism, synthesis, and storage pathways in birds can be dysregulated by only brief exposure to MeHg, with potentially significant consequences for migratory performance.

Liver Fatty Acid‐Binding Protein (LFABP) Ablation Drives Hyperplastic Expansion of Subcutaneous Adipose Tissue in Male Mice Fed High‐Fat Diet

Liver fatty acid‐binding protein (LFABP) functions both in intracellular lipid trafficking in liver and small intestine, and in systemic energy homeostasis regulation. We found that despite the marked obesity of high fat‐fed LFABP null (LFABP‐/‐) mice compared to their wild‐type (WT) counterparts, they display a ‘metabolically healthy obese’ (MHO) phenotype characterized by normoglycemia and normoinsulinemia, increased spontaneous physical activity, and a resistance to high‐fat diet (HFD)‐induced decline in exercise capacity. They also display increased muscle fatty acid oxidation suggesting that adipose tissue lipolysis is enhanced. To gain insight into the effects of Lfabp ablation on the quality of adipose tissue, we determined the mass and cellularity of the inguinal (iWAT) and epididymal (eWAT) white adipose tissues.LFABP‐/‐ mice fed HFD for 12 weeks weighed 23.7% more (BW, g: 41.7 LFABP‐/‐ vs 33.7 WT, p=0.006, n=5) and their fat pads tended to be heavier (iWAT, g: 1.1 LFABP‐/‐ vs 0.8 WT, p=0.060, n=5; eWAT, g: 1.4 LFABP‐/‐ vs 1.8 WT, p=0.065, n=5). Intriguingly, as shown in Figure 1, adipocyte size of iWAT adipocytes is smaller than that of the WT mice (44.6pL LFABP‐/‐ vs 188.1pL WT per cell, p=0.009, n=5). Adipocyte number was substantially increased in the iWAT of LFABP‐/‐ (23.6 LFABP‐/‐ vs 4.6 WT x 106, p=0.001, n=5). In contrast, the eWAT adipocyte size in the LFABP‐/‐ is comparable to WT (210pL LFABP‐/‐ vs 268 pL WT per cell, p=0.293, n=5), and eWAT fat cell number was highly variable and trended higher in the LFABP‐/‐ mice (9.8 LFABP‐/‐ vs 4.7 WT x 106, p=0.059, n=5).Because LFABP is not expressed in adipose tissue, our data suggest that its ablation promotes interorgan signaling that may limit hypertrophy and drive hyperplasia in expansion of potentially metabolically beneficial subcutaneous iWAT. More studies are warranted to elucidate the marked differences in cell size and the apparent depot‐dependent effects of LFABP‐/‐ deficiency, as well as the mechanisms by which it modulates adipocyte growth and function.

Lactiplantibacillus plantarum fermented barley extracts ameliorate high-fat-diet-induced muscle dysfunction via mitophagy.

A reduced level of fatty acid oxidation (FAO) by skeletal muscle leads to the accumulation of intermuscular fat (IMF), which is linked to impaired exercise capacity. Previously, we have reported that Lactiplantibacillus plantarum fermented barley extract (LFBE) has effective anti-obesity properties. In this study, the effects of LFBE on muscle were investigated. LFBE improved running endurance and muscle strength, which was caused by the elevation of FAO in muscle. In addition, LFBE renovated muscle regeneration through the upregulation of paired box 7 and myogenic differentiation 1 expression avoiding the injury of skeletal muscle fibers. Furthermore, total polyphenol isolated from LFBE (FTP) reinforced mobility and showed a significant protective effect on maintaining muscle fiber morphogenesis in Caenorhabditis elegans. Transmission electron microscope observation suggested FTP induced mitophagy in C.elegans body wall muscle, which was strongly connected with enhanced FAO in mitochondria. Our findings highlighted the beneficial bioactivities of FTP and its potential application for stimulating mitophagy and muscle function in obese individuals. © 2022 Society of Chemical Industry.

The effects of diet and chronic exercise on skeletal muscle ghrelin response

BackgroundRecent findings indicate that ghrelin, particularly the unacylated form (UnAG), acutely stimulates skeletal muscle fatty acid oxidation (FAO) and can preserve insulin signaling and insulin-stimulated glucose uptake in the presence of high concentrations of saturated fatty acids. However, we recently reported that the stimulatory effect of ghrelin on FAO and subsequent ability to protect insulin stimulated glucose uptake was lost following 6-weeks (6w) of chronic high fat feeding. In the current study we examined the effects of both short-term 5 day (5d) and chronic 6w high-fat diet (HFD) on muscle ghrelin response, and whether exercise training could prevent the development of muscle ghrelin resistance with 6w of HFD Methods and ResultsSoleus muscle strips were isolated from male rats to determine the direct effects of acylated (AG) and UnAG isoforms on FAO and glucose uptake. A 5d HFD did not alter the response of soleus muscle to AG or UnAG. Conversely, 6w of HFD was associated with a loss of ghrelin's ability to stimulate FAO and protect insulin stimulated glucose uptake. Muscle response to UnAG remained intact following the 6w HFD with chronic exercise training. Unexpectedly, muscle response to both AG and UnAG was also lost after 6w of low-fat diet (LFD) consumption. Protein content of the classic ghrelin receptor, GHS-R1a, was not affected by diet or training. Corticotropin-releasing hormone receptor-2 (CRF-2R) content, a putative receptor for ghrelin in muscle, was significantly decreased in soleus from 6w HFD-fed animals and increased following exercise training. This may explain the protection of UnAG response with training in HFD-fed rats but does not explain why ghrelin response was also lost in LFD-fed animals. ConclusionsUnAG protects muscle glucose uptake during acute lipid oversupply, likely due to its ability to stimulate FAO. This effect is lost in 6w HFD-fed animals but protected with exercise training. Unexpectedly, ghrelin response was lost in 6w LFD-fed animals. The loss of ghrelin response in muscle with a LFD cannot be explained by a change in putative ghrelin receptor content. We believe that the sedentary nature of the animals is a major factor in the development of muscle ghrelin resistance and warrants further research.

The aqueous fraction of Castanea crenata inner shell extract reduces obesity and intramuscular lipid accumulation via induction of mitochondrial respiration and fatty acid oxidation in muscle

BackgroundSkeletal muscle is responsible for free fatty acid (FFA) disposal via mitochondrial respiration and fatty acid oxidation (FAO). Obesity triggers high levels of circulating FFAs, which can cause intramuscular lipid (IMCL) deposition. Diverse phytochemicals, including crude Castanea crenata inner shell extract (CCE), have been shown to possess an anti-obesity effect. PurposeWe aimed to demonstrate whether the aqueous fraction of CCE (ACCE) provides an anti-obesity effect with a decrease in plasma FFAs and reduces IMCL. MethodsHigh-fat-fed C57BL/6 mice received ACCE via water intake. A204 cells incubated with fatty acids were treated with ACCE. Lipid accumulation and mitochondrial metabolism were assessed using histological and molecular techniques. ResultsACCE possessed a notably higher gallic acid content than CCE among the constituents. ACCE-administered mice exhibited reduced plasma FFA levels, adiposity, and IMCL. Muscle lipotoxicity was suppressed, including apoptosis, ER stress, and inflammation. The anti-lipid effect of ACCE was observed with the induction of mitochondrial respiration and fatty acid oxidation in muscle. ConclusionsACCE increases mitochondrial respiration and FAO in skeletal muscle and protects muscle from IMCL and lipotoxicity, reducing plasma FFA and adiposity.

Sex differences in endurance exercise capacity and skeletal muscle lipid metabolism in mice.

Previous studies suggest that sex differences in lipid metabolism exist with females demonstrating a higher utilization of lipids during exercise, which is mediated partly by increased utilization of muscle triglycerides. However, whether these changes in lipid metabolism contribute directly to endurance exercise performance is unclear. Therefore, the objective of this study was to investigate the contribution of exercise substrate metabolism to sex differences in endurance exercise capacity (EEC) in mice. Male and female C57BL/6‐NCrl mice were subjected to an EEC test until exhaustion on a motorized treadmill. The treadmill was set at a 10% incline, and the speed gradually increased from 10.2 m/min to 22.2 m/min at fixed intervals for up to 2.5 h. Tissues and blood were harvested in mice immediately following the EEC. A cohort of sedentary, non‐exercised male and female mice were used as controls. Females outperformed males by ~25% on the EEC. Serum levels of both fatty acids and ketone bodies were ~50% higher in females at the end of the EEC. In sedentary female mice, skeletal muscle triglyceride content was significantly greater compared to sedentary males. Gene expression analysis demonstrated that genes involved in skeletal muscle fatty acid oxidation were significantly higher in females with no changes in genes associated with glucose uptake or ketone body oxidation. The findings suggest that female mice have a higher endurance exercise capacity and a greater ability to mobilize and utilize fatty acids for energy.

Regulation of melanocortin-5 receptor pharmacology by two isoforms of MRAP2 in ricefield eel (Monopterus albus)

The melanocortin-5 receptor (MC5R) has been implicated in the regulation of exocrine gland secretion, immune regulation, and muscle fatty acid oxidation in mammals. Melanocortin-2 receptor accessory protein 2 (MRAP2) can modulate trafficking, ligand binding, and signaling of melanocortin receptors. To explore potential interaction between ricefield eel (Monopterus albus) MC5R and MRAP2s (maMC5R, maMRAP2X1, and maMRAP2X2), herein we studied the pharmacological characteristics of maMC5R and its modulation by maMRAP2s expressed in the human embryonic kidney cells. Three agonists, α-melanocyte-stimulating hormone (α-MSH), ACTH (1–24), and [Nle4, D-Phe7]-α-MSH, could bind to maMC5R and induce intracellular cAMP production dose-dependently. Compared with human MC5R (hMC5R), maMC5R displayed decreased maximal binding but higher binding affinity to α-MSH or ACTH (1–24). When stimulated with α-MSH or ACTH (1–24), maMC5R showed significantly lower EC50 and maximal response than hMC5R. Two maMRAP2s had no effect on cell surface expression of maMC5R, whereas they significantly increased maximal binding. Only maMRAP2X2 significantly decreased the binding affinity of ACTH (1–24). Both maMRAP2X1 and maMRAP2X2 significantly reduced maMC5R efficacy but did not affect ligand sensitivity. The availability of maMC5R pharmacological characteristics and modulation by maMRAP2s will assist the investigation of its roles in regulating diverse physiological processes in ricefield eel.

Maternal High-Fat Diet Increases Fetal Muscle Fat Metabolism and Fatty Acid Transporter Expression in an Ovine Model

ObjectivesExcessive maternal dietary fat consumption during pregnancy may be linked to adverse effects on offspring health, including greater risk of developing metabolic syndrome later in life. Metabolic syndrome is generally considered to be a preventable condition, but the extent to which it is “programmed” during fetal development remains unclear. The aim of this study was to determine the effect of a maternal high-fat diet (MHFD) during pregnancy on fetal muscle oxidative metabolism and related protein and mRNA expressions in an ovine model. MethodsWhite-faced ewes were fed either a control diet (Show-rite NewCo Lamb Feed-17% protein, 5% Fat) or a high-fat diet (Show-rite NewCo Lamb Feed + 6% Rumen-protected Fat) from 2–3 weeks before pregnancy until mid-gestation (75 days), when a C-section was performed to collect the placenta and fetal tissues for analysis. ResultsMHFD tended to increase fetal body and organ weights, but only significantly increased fetal body length and liver mass (P < 0.05). MHFD increased mRNA expression of placental (cotyledon) fatty acid transport protein-1 (FATP-1) and peroxisome proliferator activated receptor gamma, suggesting an upregulation of placental fatty acid metabolism and transport. Fetal muscle fatty acid oxidation capacity was greater in animals from MHFD pregnancies, with no effect on pyruvate oxidation. This was associated with greater fetal muscle mRNA and protein expression of FATP4, while mRNA expression glucose transporters (GLUT1 and GLUT3) decreased. Muscle expression of insulin signaling enzymes reflected a mild decreases in insulin sensitivity, but these did not reach statistical significance. ConclusionsThese studies indicate that MHFD induces an increase in placental and fetal muscle fatty acid transport and oxidation capacity, and favors lower blood glucose uptake compared to controls. Whether these shifts in fetal metabolism predispose offspring from MHFD pregnancies to elevated blood sugar and Type 2 diabetes later in life merits further investigation. Funding SourcesColorado Agricultural Experiment Station.