Intramuscular Lipid Research Articles

The effects of a protein enriched diet with lean red meat combined with a multi-modal exercise program on muscle and cognitive health and function in older adults: study protocol for a randomised controlled trial.

BackgroundAge-related muscle wasting has been strongly implicated with falls and fractures in the elderly, but it has also been associated with cognitive decline and dementia. Progressive resistance training (PRT) and adequate dietary protein are recognised as important contributors to the maintenance of muscle health and function in older adults. However, both factors also have the potential to improve brain function and prevent cognitive decline via several pathways, including the regulation of various growth and neurotrophic factors [insulin-like growth factor-1 (IGF-1)]; brain-derived growth factor (BDNF)] and/or the modulation of systemic inflammation. The primary aim of this study is to investigate whether a modest increase in dietary protein achieved through the consumption of lean red meat three days per week, when combined with PRT, can enhance muscle mass, size and strength and cognitive function in community-dwelling older people.Methods/DesignThe study design is a 48-week randomised controlled trial consisting of a 24-week intervention with a 24-week follow-up. Men and women (n=152) aged 65 years and over residing in the community will be randomly allocated to: 1) PRT and provided with 220 g (raw weight) of lean red meat to be cooked and divided into two 80 g servings on each of the three days that they complete their exercise session, or 2) control PRT in which participants will be provided with and advised to consume ≥1 serving (~1/2 cup) of rice and/or pasta or 1 medium potato on each of the three training days. The primary outcome measures will be muscle mass, size and strength and cognitive function. Secondary outcomes will include changes in: muscle function, neural health (corticospinal excitability and inhibition and voluntary activation), serum IGF-1 and BDNF, adipokines and inflammatory markers, fat mass and inter-/intra-muscular fat, blood pressure, lipids and health-related quality of life. All outcome measures will be assessed at baseline and 24 weeks, with the exception of cognitive function and the various neurobiological and inflammatory markers which will also be assessed at week 12.DiscussionThe findings from this study will provide important new information on whether a modest increase in dietary protein achieved through the ingestion of lean red meat can enhance the effects of PRT on muscle mass, size and strength as well as cognitive function in community-dwelling older adults. If successful, the findings will form the basis for more precise exercise and nutrition guidelines for the management and prevention of age-related changes in muscle and neural health and cognitive function in the elderly.Trial registrationAustralian New Zealand Clinical Trials Registry: ACTRN12613001153707. Date registered 16th October, 2013.

Effect of tamoxifen on fatty degeneration and atrophy of rotator cuff muscles in chronic rotator cuff tear: An animal model study.

Fatty degeneration of the rotator cuff muscles is an irreversible change resulting from chronic rotator cuff tear and is associated with poor clinical outcomes following rotator cuff repair. We evaluated the effect of Tamoxifen, a competitive estrogen receptor inhibitor, on fatty degeneration using a mouse model for chronic rotator cuff tear. Sixteen adult mice were divided into two diet groups (Tamoxifen vs. Regular) and subjected to surgical creation of a large rotator cuff tear and suprascapular nerve transection in their left shoulder with the right shoulder serving as a control. The rotator cuff muscles were harvested at 16 weeks and subjected to histology and RT-PCR for adipogenic and myogenic markers. Histology showed substantially decreased atrophy and endomysial inflammation in Tamoxifen group, but no significant differences in the amount of intramuscular adipocytes and lipid droplets compared to the Regular group. With RT-PCR, the operated shoulders showed significant upregulation of myogenin and PPAR-γ, and downregulation of myostatin compared to the nonsurgical shoulder. No significant differences of gene expression were found between the two diet groups. Our study demonstrated that tamoxifen diet leads to decreased muscle atrophy and inflammatory changes following chronic rotator cuff tear, but has no apparent effect on adipogenesis.

Effects of Choline on Meat Quality and Intramuscular Fat in Intrauterine Growth Retardation Pigs.

The aim of this study was to investigate the effects of choline supplementation on intramuscular fat (IMF) and lipid oxidation in IUGR pigs. Twelve normal body weight (NBW) and twelve intrauterine growth retardation (IUGR) newborn piglets were collected and distributed into 4 treatments (Normal: N, Normal+Choline: N+C, IUGR: I, and IUGR+Choline: I+C) with 6 piglets in each treatment. At 23 d of age, NBW and IUGR pigs were fed basal or choline supplemented diets. The results showed that the IUGR pigs had significantly lower (P<0.05) BW as compared with the NBW pigs at 23 d, 73 d, and 120 d of age, however, there was a slight decreased (P>0.05) in BW of IUGR pigs than the NBW pigs at 200 d. Compared with the NBW pigs, pH of meat longissimus dorsi muscle was significantly lower (P<0.05), and the meat color was improved in IUGR pigs. The malondialdehyde (MDA) levels were significantly decreased (P<0.05), while triglyceride (TG) and IMF contents were significantly higher (P<0.05) in the IUGR pigs than the NBW pigs. IUGR up-regulated the mRNA gene expression of fatty acid synthetase (FAS) and acetyl-CoA carboxylase (ACC). Dietary choline significantly increased (P<0.05) the BW at 120d of age, however, significantly decreased (P<0.05) the TG and IMF contents in both IUGR and NBW pigs. FAS and sterol regulatory element-binding proteins 1 (SREBP1) mRNA gene expressions were increased (P<0.05) while the muscle-carnitine palmityl transferase (M-CPT) and peroxisome proliferators-activated receptorγ (PPARγ) mRNA (P<0.05) gene expressions were decreased in the muscles of the IUGR pigs by choline supplementation. Furthermore, choline supplementation significantly increased (P<0.05) the MDA content as well as the O2•¯ scavenging activity in meat of IUGR pigs. The results suggested that IUGR pigs showed a permanent stunting effect on the growth performance, increased fat deposition and oxidative stress in muscles. However, dietary supplementation of choline improved the fat deposition via enhancing the lipogenesis and reducing the lipolysis.

Effects of high pressure in combination with thermal treatment on lipid hydrolysis and oxidation in pork

For investigating the effects of high pressure in combination with thermal (HP-T) treatment on lipid hydrolysis and oxidation in pork, minced pork was treated with combinations of different pressures (200, 400, and 600 MPa) and temperatures (20 and 50 °C); the content and fatty acid profiles of intramuscular lipids, peroxide value (POV), thiobarbituric acid reactive substances (TBARS), and lipoxygenase (LOX) activity were measured. Treatment of samples at pressures of 400 MPa or above significantly enhanced the uneven hydrolysis of phospholipids, resulting in more polyunsaturated fatty acids (PUFA) in degraded phospholipids or generated free fatty acids (FFA); treatment temperature had a synergistic effect on phospholipid hydrolysis. Both POV and TBARS values increased with the intensity of treatment conditions until they reached the highest value, and then gradually decreased; neither corresponded closely with the changes in LOX activity. ANOVA-partial least squares regression (APLSR) analysis indicated that phospholipid hydrolysis positively correlated with TBARS but weakly correlated with POV. Thus, phospholipid hydrolysis could enhance lipid secondary oxidation, which mainly arising from auto-oxidation.

Lipid‐Induced Anabolic Resistance in Skeletal Muscle is Associated with Inflammation in Aging, but Not Obesity

The loss of skeletal muscle mass is observed in many pathophysiological conditions including, aging and obesity. Chronic inflammation is observed with aging and obesity may play a critical role in the muscle's resistance to anabolic stimulation. A characteristic metabolic feature of both these conditions is an increase in the accumulation of intramyocellular lipid (IMCL) content. Therefore, we tested the hypothesis that lipid-induced anabolic resistance is mediated by inflammation in both aging and obesity. In order to test this hypothesis we used C57BL/6 mice aged 6 months and fed a control diet (YNG, 15% fat/kcal) or a high-fat (FAT, 60% fat/kcal) and mice aged 24 months fed a control diet (OLD) for 12 wk. The accumulation of intramuscular lipids was increased in both OLD and FAT compared to YNG. The phosphorylation of anabolic signaling components was increased in all groups after insulin stimulation but differentially affected in OLD and FAT. Differences in the activation and gene expression of the NFκB proinflammatory signaling pathway were only observed in OLD, but not FAT. We conclude that the accumulation of lipids during aging and obesity have divergent effects on inflammation in skeletal muscle.

Changes in mitochondrial perilipin 3 and perilipin 5 protein content in rat skeletal muscle following endurance training and acute stimulated contraction.

What is the central question of this study? The aim was to determine whether mitochondrial protein content of perilipin 3 (PLIN3) and perilipin 5 (PLIN5) is increased following endurance training and whether mitochondrial PLIN5 protein is increased to a greater extent in endurance-trained rats when compared with sedentary rats following acute contraction. What is the main finding and its importance? Mitochondrial PLIN3 but not PLIN5 protein was increased in endurance-trained compared with sedentary rats, suggesting a mitochondrial role for PLIN3 due to chronic exercise. Contrary to our hypothesis, acute mitochondrial PLIN5 protein was similar in both sedentary and endurance-trained rats. Endurance training results in an increased association between skeletal muscle lipid droplets and mitochondria. This association is likely to be important for the expected increase in intramuscular fatty acid oxidation that occurs with endurance training. The perilipin family of lipid droplet proteins, PLIN(2-5), are thought to play a role in skeletal muscle lipolysis. Recently, results from our laboratory demonstrated that skeletal muscle mitochondria contain PLIN3 and PLIN5 protein. Furthermore, 30min of stimulated contraction induces an increased mitochondrial PLIN5 content. To determine whether mitochondrial content of PLIN3 and PLIN5 is altered with endurance training, Sprague-Dawley rats were randomized into sedentary or endurance-trained groups for 8weeks of treadmill running followed by an acute (30min) sciatic nerve stimulation to induce lipolysis. Mitochondrial PLIN3 protein was ∼1.5-fold higher in red gastrocnemius of endurance-trained rats compared with sedentary animals, with no change in mitochondrial PLIN5 protein. In addition, there was an increase in plantaris intramuscular lipid storage. Acute electrically stimulated contraction in red gastrocnemius from sedentary and endurance-trained rats resulted in a similar increase of mitochondrial PLIN5 between these two groups, with no net change in PLIN3 in either group. Plantaris intramuscular lipid content decreased to a similar extent in sedentary and endurance-trained rats. These results suggest that while total mitochondrial PLIN5 content is not altered by endurance training, PLIN5 does have an acute role in the mitochondrial fraction during muscle contraction. Conversely, mitochondrial PLIN3 does not change acutely with muscle contraction, but PLIN3 content was increased following endurance training, indicating a role in chronic adaptations of skeletal muscle.

Effect of Growth on Fatty Acid Composition of Total Intramuscular Lipid and Phospholipids in Ira Rabbits.

The changes in fatty acid composition of total intramuscular lipid and phospholipids were investigated in the longissimus dorsi, left-hind leg muscle, and abdominal muscle of male Ira rabbits. Changes were monitored at 35, 45, 60, 75, and 90 d. Analysis using gas chromatography identified 21 types of fatty acids. Results showed that the intramuscular lipid increased and the intramuscular phospholipids (total intramuscular lipid %) decreased in all muscles with increasing age (p<0.05). An abundant amount of unsaturated fatty acids, especially polyunsaturated fatty acids, was distributed in male Ira rabbits at different ages and muscles. Palmitic acid (C16:0), stearic acid (C18:0), oleic acid (C18:1), linoleic acid (C18:2), and arachidonic acid (C20:4) were the major fatty acids, which account to the dynamic changes of the n-6/n-3 value in Ira rabbit meat.

18-Carbon polyunsaturated fatty acids ameliorate palmitate-induced inflammation and insulin resistance in mouse C2C12 myotubes

Skeletal muscle is a major site of insulin action. Intramuscular lipid accumulation results in inflammation, which has a strong correlation with skeletal muscle insulin resistance (IR). The aim of this study was to explore the effects of linoleic acid, alpha-linolenic acid, and gamma-linolenic acid (GLA), 18-carbon polyunsaturated fatty acids (PUFAs), on palmitic acid (PA)-induced inflammatory responses and IR in C2C12 myotubes. Our data demonstrated that these three test 18-carbon PUFAs can inhibit PA-induced interleukin-6 and tumor necrosis factor-α messenger RNA (mRNA) expression and IR as evidenced by increases in phosphorylated AKT and the 160-kD AKT substrate, mRNA and plasma membrane protein expression of glucose transporter 4, and glucose uptake. Moreover, the 18-carbon PUFAs blocked the effects of PA on activation of mitogen-activated protein kinases (MAPKs), protein kinase C-θ (PKC-θ), AMP-activated protein kinase (AMPK) and nuclear factor-κB (NF-κB). Of note, supplementation with GLA-rich borage oil decreased proinflammatory cytokine production and hindered the activation of MAPKs, PKC-θ and NF-κB in the skeletal muscles of diabetic mice. The 18-carbon PUFAs did not reverse PA-induced inflammation or IR in C2C12 myotubes transfected with a constitutively active mutant IκB kinase-β plasmid, which suggests the importance of the inhibition of NF-κB activation by the 18-carbon PUFAs. Moreover, blockade of AMPK activation by short hairpin RNA annulled the inhibitory effects of the 18-carbon PUFAs on PA-induced IR but not inflammation. Our findings suggest that the 18-carbon PUFAs may be useful in the management of PA-induced inflammation and IR in myotubes.

Rearing system and oleic acid supplementation effect on carcass and lipid characteristics of two muscles from an obese pig breed

Quality of pork depends on genotype, rearing and pre- and post-slaughter conditions. However, no information is available on rearing system changes and oleic acid supplementation on carcass characteristics and fatty acid (FA) profile of pork from the Alentejano (AL) pig, an obese breed. This study evaluates the effects of feeding low (LO) or high oleic acid diets (HO) to AL pigs reared in individual pens (IND) or outdoor (OUT) with access to pasture. Carcass composition was obtained and longissimus dorsi and semimembranosus samples were collected to analyse chemical composition and neutral and polar intramuscular lipids FA profile by gas chromatography. Statistical analysis was performed by a two-way ANOVA for rearing system and diet effects. OUT-reared pigs presented leaner carcasses than IND-reared ones. Both muscles presented lower intramuscular lipid content in OUT-reared pigs. Treatments affected the FA profile of muscles. Overall, OUT-reared pigs presented lower n-6/n-3 FA ratios, whereas pigs fed the HO diet exhibited lower saturated fatty acids (SFA), higher monounsaturated fatty acids (MUFA) levels and lower thrombogenic indexes on neutral intramuscular lipids than LO-fed pigs. On the polar fraction, OUT-reared pigs presented lower SAT and n-6/n-3 FA ratio, and higher polyunsaturated fatty acids (PUFA) levels on both muscles. Pigs fed the HO diet exhibited higher MUFA and lower PUFA levels on both muscles, and lower SAT levels on semimembranosus. This study shows rearing system and oleic acid supplementation have complementary effects and influence carcass composition and the nutritional quality of meat.

Effects of Aerobic Exercise on the Intramuscular Lipid and Glycogen Content of Fiber Types in Soleus Muscles of Non-Alcoholic Steatohepatitis Model Rats

We studied the effects of exercise on muscle mitochondria, and lipid and glycogen content in non-alcoholic steatohepatitis (NASH) model rats. Male Sprague-Dawley rats were randomly separated into 3 groups: the control group was fed standard chow; the NASH group was fed a methionine-choline-deficient high-fat diet (MCD); the NASH-exercise group was fed the MCD and exercised three times a week. Exercise training consisted of continuous running for thirty minutes at a 13 m/min, 6° slope on a motor-driven rodent treadmill for 6 weeks. Mitochondria content in NASH group decreased in the both fiber types compared with those of the control group. As compared between the NASH and NASH-exercise groups, however, exercise not only promoted significant improvements in liver fibrosis and cirrhosis and triglyceride (TG) content but also increased mitochondria content in type I muscle fiber in particular. These data suggest that exercise improved hepatic steatosis in NASH model rats and can prevent the progression of NASH.

Skeletal muscle phenotype affects fasting-induced mitochondrial oxidative phosphorylation flexibility in cold-acclimated ducklings.

Starvation is particularly challenging for endotherms that remain active in cold environments or during winter. The aim of this study was to determine whether fasting-induced mitochondrial coupling flexibility depends upon the phenotype of skeletal muscles. The rates of oxidative phosphorylation and mitochondrial efficiency were measured in pectoralis (glycolytic) and gastrocnemius (oxidative) muscles from cold-acclimated ducklings (Cairina moschata). Pyruvate and palmitoyl-l-carnitine were used in the presence of malate as respiratory substrates. Plasma metabolites, skeletal muscle concentrations of triglycerides, glycogen and total protein and mitochondrial levels of oxidative phosphorylation complexes were also quantified. Results from ad libitum fed ducklings were compared with those from ducklings that were fasted for 4 days. During the 4 days of nutritional treatment, birds remained in the cold, at 4°C. The 4 days of starvation preferentially affected the pectoralis muscles, inducing an up-regulation of mitochondrial efficiency, which was associated with a reduction of both total muscle and mitochondrial oxidative phosphorylation protein, and with an increase of intramuscular lipid concentration. By contrast, fasting decreased the activity of oxidative phosphorylation but did not alter the coupling efficiency and protein expression of mitochondria isolated from the gastrocnemius muscles. Hence, the adjustment of mitochondrial efficiency to fasting depends upon the muscle phenotype of cold-acclimated birds. Furthermore, these results suggest that the reduced cost of mitochondrial ATP production in pectoralis muscles may trigger lipid storage within this tissue and help to sustain an important metabolic homeostatic function of skeletal muscles, which is to maintain levels of amino acids in the circulation during the fast.

Multi-state consumer acceptance of porkloin chops of varied intramuscular lipid content

Multi-state consumer acceptance of porkloin chops of varied intramuscular lipid content

Short-term muscle disuse atrophy is not associated with increased intramuscular lipid deposition or a decline in the maximal activity of key mitochondrial enzymes in young and older males

Aging is generally accompanied by a progressive loss of skeletal muscle mass and impairments in metabolic function. Even a few days of muscle disuse (such as that occurring during injury or illness) leads to considerable loss of muscle mass and strength. It has been speculated that short, successive periods of muscle disuse throughout the lifespan may be largely responsible for the age-related loss of muscle mass. However, it remains unknown whether such short periods of disuse also induce impairments in metabolic function within skeletal muscle. Here, we investigated the effects of a five day period of muscle disuse on intramyocellular triacylglycerol (IMTG) content, muscle oxidative capacity, and the expression of key genes that regulate oxidative metabolism in healthy young and elderly men. Muscle biopsies were collected from healthy, young (n=12; 23±1y) and elderly (n=12; 70±1y) men prior to and immediately after a five day period of one-legged knee immobilization by way of a full leg cast. At baseline, elderly men had a greater IMTG content when compared with the young (56.2±5.1 and 34.8±7.3μmol·g−1, respectively; P<0.05) with no changes in either group following immobilization (53.4±5.0 and 35.7±5.0μmol·g−1, respectively; P>0.05). In line, five days of disuse did not lower citrate synthase, β-HAD or cytochrome C oxidase activity in skeletal muscle tissue. Pyruvate dehydrogenase activity increased following immobilization in the older subjects only, from 0.39±0.06 to 0.55 0.05μmol·g−1·min−1 (71±33%; P<0.01). The skeletal muscle mRNA expression of PGC1α and citrate synthase both declined following immobilization in both the young and elderly subjects. We conclude that five days of muscle disuse does not increase intramuscular lipid deposition or reduce the maximal activity of key mitochondrial enzymes within the skeletal muscle of young or older men.

Effects of the porcine IGF2 intron 3-G3072A mutation on carcass cutability, meat quality, and bacon processing.

A SNP in a regulatory region of intron 3 within the porcine IGF2 gene (IGF2-G3072A) is associated with increased lean deposition and decreased fat deposition in pigs with paternal A alleles (APat) compared with pigs with paternal G alleles (GPat). However, data regarding fresh and processed meat quality characteristics of pigs with different alleles for this polymorphism are limited. A single heterozygote (AG) boar was bred to homozygous (AA) commercial Yorkshire-cross sows producing F1 barrows and gilts with either GPat or APat. Two farrowing groups of barrows and gilts were group housed, provided ad libitum access to a diet that met or exceeded NRC nutrient recommendations throughout production, and slaughtered at 176 d (±4 d) of age. Fresh LM quality and estimated percent fat-free lean measurements were taken on the left side of carcasses, while carcass cutouts were completed with right sides. Fresh belly and bacon processing traits were characterized for only block 1 pigs. Pig was treated as the experimental unit for all analyses. Ending live weight and HCW were not affected by IGF2 allele; however, 10th rib backfat thickness was 0.41 cm less (P=0.01), loin eye area was 4.0 cm2 greater (P=0.01), and predicted fat-free lean was over 2 percentage units greater (P<0.01) in APat pigs compared with GPat pigs. Furthermore, boneless lean cuts from the shoulder, loin, and ham were heavier (P<0.05) in APat pigs compared with GPat pigs. Minolta L* value was 2.36 units greater (P=0.03) but cooking loss was 1.82 percentage units greater (P<0.01) in APat pigs compared with GPat pigs. Additionally, despite reductions in subcutaneous fat, extractable intramuscular lipid from the LM was 0.64 percentage units greater (P=0.02) in APat pigs compared with GPat pigs. Bellies were 7.17 mm thinner (P=0.01), had 7.27 cm less flop distance (P=0.05), and tended to have 1.34 units greater iodine value (P=0.09) in APat pigs compared with GPat pigs. While not statistically different (P=0.30), the magnitude of difference in slicing yield as a percentage of green weight was 1.57 percentage units between bellies from APat pigs (85.83%) and bellies from GPat pigs (87.40%). Pigs with GPat had superior belly quality that may positively impact commercial bacon production. However, pigs with APat yielded a greater amount of lean product at the expense of producing lighter LM color and increased cooking loss.

Intermuscular and intramuscular adipose tissues: Bad vs. good adipose tissues

Human studies of the influence of aging and other factors on intermuscular fat (INTMF) were reviewed. Intermuscular fat increased with weight loss, weight gain, or with no weight change with age in humans. An increase in INTMF represents a similar threat to type 2 diabetes and insulin resistance as does visceral adipose tissue (VAT). Studies of INTMF in animals covered topics such as quantitative deposition and genetic relationships with other fat depots. The relationship between leanness and higher proportions of INTMF fat in pigs was not observed in human studies and was not corroborated by other pig studies. In humans, changes in muscle mass, strength and quality are associated with INTMF accretion with aging. Gene expression profiling and intrinsic methylation differences in pigs demonstrated that INTMF and VAT are primarily associated with inflammatory and immune processes. It seems that in the pig and humans, INTMF and VAT share a similar pattern of distribution and a similar association of components dictating insulin sensitivity. Studies on intramuscular (IM) adipocyte development in meat animals were reviewed. Gene expression analysis and genetic analysis have identified candidate genes involved in IM adipocyte development. Intramuscular (IM) adipocyte development in human muscle is only seen during aging and some pathological circumstance. Several genetic links between human and meat animal adipogenesis have been identified. In pigs, the Lipin1 and Lipin 2 gene have strong genetic effects on IM accumulation. Lipin1 deficiency results in immature adipocyte development in human lipodystrophy. In humans, overexpression of Perilipin 2 (PLIN2) facilitates intramyocellular lipid accretion whereas in pigs PLIN2 gene expression is associated with IM deposition. Lipins and perilipins may influence intramuscular lipid regardless of species.

Higher PLIN5 but not PLIN3 content in isolated skeletal muscle mitochondria following acute in vivo contraction in rat hindlimb

Contraction‐mediated lipolysis increases the association of lipid droplets and mitochondria, indicating an important role in the passage of fatty acids from lipid droplets to mitochondria in skeletal muscle. PLIN3 and PLIN5 are of particular interest to the lipid droplet–mitochondria interaction because PLIN3 is able to move about within cells and PLIN5 associates with skeletal muscle mitochondria. This study primarily investigated: 1) if PLIN3 is detected in skeletal muscle mitochondrial fraction; and 2) if mitochondrial protein content of PLIN3 and/or PLIN5 changes following stimulated contraction. A secondary aim was to determine if PLIN3 and PLIN5 associate and whether this changes following contraction. Male Long Evans rats (n = 21; age, 52 days; weight = 317 ± 6 g) underwent 30 min of hindlimb stimulation (10 msec impulses, 100 Hz/3 sec at 10–20 V; train duration 100 msec). Contraction induced a ~50% reduction in intramuscular lipid content measured by oil red‐O staining of red gastrocnemius muscle. Mitochondria were isolated from red gastrocnemius muscle by differential centrifugation and proteins were detected by western blotting. Mitochondrial PLIN5 content was ~1.6‐fold higher following 30 min of contraction and PLIN3 content was detected in the mitochondrial fraction, and unchanged following contraction. An association between PLIN3 and PLIN5 was observed and remained unaltered following contraction. PLIN5 may play a role in mitochondria during lipolysis, which is consistent with a role in facilitating/regulating mitochondrial fatty acid oxidation. PLIN3 and PLIN5 may be working together on the lipid droplet and mitochondria during contraction‐induced lipolysis.

Abstract 556: Obese Diabetic Mice Exhibited Severe Intramuscular Lipid Accumulation after Muscle Injury

Background: Sarcopenic obesity, a loss of muscle and a concomitant increase in ectopic fat, is reported to increase in life-style related diseases. Especially the prevalence of sarcopenia is greater in patients with type 2 diabetes mellitus (T2DM) than in non-diabetic subjects. Increase in fat filtration enhances insulin resistance and is associated with metabolic risk factors. However, the detailed mechanism of such ectopic fat filtration in muscle of T2DM patients has not been well investigated. Here, we assessed muscle regeneration using muscle-injured models in obese diabetic mice. Methods: Male eight-week-old wild-type mice (C57BL6) and T2DM mice (KKAy) were undergone intramuscular injection of cardiotoxin (Ctx) (100uL/10uM) into tibia. After two weeks, muscle were removed and stained with hematoxylin and eosin. KKAy replaced by bone marrow prepared from GFP-transgenic mice were generated by 8 Gy whole body X-ray irradiation. Cell-differentiation was evaluated by immunofluorescent analysis. Some mice were treated with all trans-retinoic acid (ATRA) and PPAR-gamma antagonist, GW9662. Results: In 26 week-old KKAy, magnetic resonance imaging analysis showed that intramuscular fat deposition was estimated as higher intensity compared with C57BL6 mice. Treatment with Ctx showed a significant increase in honey comb structure which was perilipin positive. KKAy mice exhibited impaired muscle regeneration. Tibial muscle weight was approximately a half in Ctx-injured KKAy. Such change was also observed in another diabetic mouse model, db/db, but not in streptozocin-induced diabetic mice. Fat formation was remarkably increased in aged KKAy compared with younger mice. Several fat formation was GFP-positive in GFP-chimeric mice indicating that fat tissue was partially generated from marrow cells. Fat formation was desmin-negative and vimentin-positive indicating that these are mesenchymal origin. Treatment with ATRA reduced fat filtration. On the other hand, treatment with GW9662 enhanced fat filtration. Conclusion: Obese diabetic mice showed significantly impaired muscle regeneration with replacement of fat deposition, suggesting that diabetes enhanced sarcopenic obesity possibly due to inhibitory effect of cell differentiation.

Muscle ectopic fat deposition contributes to anabolic resistance in obese sarcopenic old rats through eIF2α activation.

Obesity and aging are characterized by decreased insulin sensitivity (IS) and muscle protein synthesis. Intramuscular ceramide accumulation has been implicated in insulin resistance during obesity. We aimed to measure IS, muscle ceramide level, protein synthesis, and activation of intracellular signaling pathways involved in translation initiation in male Wistar young (YR, 6-month) and old (OR, 25-month) rats receiving a low- (LFD) or a high-fat diet (HFD) for 10 weeks. A corresponding cellular approach using C2C12 myotubes treated with palmitate to induce intracellular ceramide deposition was taken. A decreased ability of adipose tissue to store lipids together with a reduced adipocyte diameter and a development of fibrosis were observed in OR after the HFD. Consequently, OR fed the HFD were insulin resistant, showed a strong increase in intramuscular ceramide level and a decrease in muscle protein synthesis associated with increased eIF2α phosphorylation. The accumulation of intramuscular lipids placed a lipid burden on mitochondria and created a disconnect between metabolic and regulating pathways in skeletal muscles of OR. In C2C12 cells, palmitate-induced ceramide accumulation was associated with a decreased protein synthesis together with upregulated eIF2α phosphorylation. In conclusion, a reduced ability to expand adipose tissues was found in OR, reflecting a lower lipid buffering capacity. Muscle mitochondrial activity was affected in OR conferring a reduced ability to oxidize fatty acids entering the muscle cell. Hence, OR were more prone to ectopic muscle lipid accumulation than YR, leading to decreased muscle protein anabolism. This metabolic change is a potential therapeutic target to counter sarcopenic obesity.

Resveratrol supplementation does not augment performance adaptations or fibre-type-specific responses to high-intensity interval training in humans.

The present study examined the effect of concurrent exercise training and daily resveratrol (RSV) supplementation (150 mg) on training-induced adaptations following low-dose high-intensity interval training (HIIT). Sixteen recreationally active (∼22 years, ∼51 mL·kg(-1)·min(-1)) men were randomly assigned in a double-blind fashion to either the RSV or placebo group with both groups performing 4 weeks of HIIT 3 days per week. Before and after training, participants had a resting muscle biopsy taken, completed a peak oxygen uptake test, a Wingate test, and a submaximal exercise test. A main effect of training (p < 0.05) and interaction effect (p < 0.05) on peak aerobic power was observed; post hoc pairwise comparisons revealed that a significant (p < 0.05) increase occurred in the placebo group only. Main effects of training (p < 0.05) were observed for both peak oxygen uptake (placebo - pretraining: 51.3 ± 1.8, post-training: 54.5 ± 1.5 mL·kg(-1)·min(-1), effect size (ES) = 0.93; RSV - pretraining: 49.6 ± 2.2, post-training: 52.3 ± 2.5 mL·kg(-1)·min(-1), ES = 0.50) and Wingate peak power (placebo: pretraining: 747 ± 39, post-training: 809 ± 31 W, ES = 0.84; RSV - pretraining: 679 ± 39, post-training: 691 ± 43 W, ES = 0.12). Fibre-type distribution was unchanged, while a main effect of training (p < 0.05) was observed for succinate dehydrogenase activity and glycogen content, but not α-glycerophosphate dehydrogenase activity or intramuscular lipids in type I and IIA fibres. The fold change in PGC-1α, SIRT1, and SOD2 gene expression following training was significantly (p < 0.05) lower in the RSV group than placebo. These results suggest that concurrent exercise training and RSV supplementation may alter the normal training response induced by low-volume HIIT.

Effect of corn supplementation of grass finishing of Holstein bulls on fatty acid composition of meat lipids.

Finishing Holstein young bulls exclusively on pasture generally results in very lean carcass and meat, but corn supplementation is expected to simultaneously improve carcass traits and intramuscular lipids (IML). The expected increase in IML would allow for a larger 18:2c9,t11 (CLA) deposition in meat without affecting the n-3 PUFA present in LM phospholipids (PL). Holstein bulls (n = 33) with initial BW of 423 ± 52.4 kg reared exclusively on pasture were assigned to 1 of 3 finishing period (85 d) diets: finished exclusively on pasture (P0) or finished on pasture and individually supplemented with 4 (P4) or 8 kg/d (P8) of ground corn. Final BW (546 ± 56.3 kg) was not affected (P > 0.05) by corn supplementation, but ADG increased (P < 0.01) with the increasing corn supplementation level from 1.23 kg/d for P0 to 1.44 kg/d for P4 and to 1.67 kg/d for P8. Subcutaneous fat depth in P0 bulls was 0.8 mm and increased (P < 0.001) in both P4 (2.9 mm) and P8 (2.7 mm) bulls, but no difference (P = 0.73) was observed between P4 and P8 bulls. Similarly, the IML increased with corn supplementation, from 1.84 g/100 g muscle in P0 to 2.96 in P4 and to 3.24 in P8, but no difference (P = 0.55) was found between P4 and P8 bulls. Corn supplementation decreased (P < 0.01) 18:1t11 in neutral lipids (NL) but not 18:2c9,t11 (P > 0.34). The 18:1t10 (mg/g total NL fatty acid [FA] ± SEM) were 2.5 ± 0.13 in P0, 5.5 ± 1.68 in P4, and 14.8 ± 3.18 in P8 bulls, being greater in P8 compared with P4 (P = 0.02). Total FA in muscle PL and SFA were unaffected, but increasing corn supplementation resulted (P < 0.001) in an increase of 18:2n-6 in PL by replacement of mostly the 18:1c9 and 18:3n-3. Notably, the total number of cis double bonds present in FA of PL remained constant (P = 0.74) with corn supplementation. Compared with P0, corn supplementation maintained (P > 0.05) the high n-3 PUFA content in meat (mg/100 g meat) and increased the 18:2c9,t11 (P = 0.028) and 18:1c9 (P < 0.001). However, increasing corn supplementation from 4 to 8 kg/d increased the 18:1t10 (P = 0.031) and had no effect on 18:2c9,t11. Therefore, supplementing grass-finished Holstein bulls with moderate amounts of ground corn (4 kg/d) increased carcass fat cover and IML, maintained n-3 PUFA, and increased 18:2c9,t11 content in meat, whereas greater corn supplementation (8 kg/d; P8) resulted in no further improvements.