l‐Citrulline Supplementation‐Increased Skeletal Muscle PGC‐1α Expression Is Associated with Exercise Performance and Increased Skeletal Muscle Weight

Scope

l ‐citrulline has recently been reported as a more effective supplement for promoting intracellular nitric oxide (NO) production compared to l ‐arginine. Here, the effect of l ‐citrulline on skeletal muscle and its influence on exercise performance were investigated. The underlying mechanism of its effect, specifically on the expression of skeletal muscle peroxisome proliferator‐activated receptor‐gamma coactivator‐1α (PGC‐1α), was also elucidated.

Methods and results

Six‐week‐old ICR mice were orally supplemented with l ‐citrulline (250 mg kg^?1) daily, and their performance in weight‐loaded swimming exercise every other day for 15 days, was evaluated. In addition, mice muscles were weighed and evaluated for the expression of PGC‐1α and PGC‐1α‐regulated genes. Mice orally supplemented with l ‐citrulline had significantly higher gastrocnemius and biceps femoris muscle mass. Although not statistically significant, l ‐citrulline prolonged the swimming time to exhaustion. PGC‐1α upregulation was associated with vascular endothelial growth factor α (VEGFα) and insulin‐like growth factor 1 (IGF‐1) upregulation. VEGFα and IGF‐1 are important for angiogenesis and muscle growth, respectively, and are regulated by PGC‐1α. Treatment with NG‐nitro‐l ‐arginine methyl ester hydrochloride (l ‐NAME), a nitric oxide synthesis inhibitor, suppressed the l ‐citrulline‐induced PGC‐1α upregulation in vitro.

Conclusion

Supplementation with l ‐citrulline upregulates skeletal muscle PGC‐1α levels resulting in higher skeletal muscle weight that improves time to exhaustion during exercise.

     

Rice protein concentrate partially replaces dried whey in the diet for early‐weaned piglets and improves their growth performance

BACKGROUND: Recently, rice protein concentrate (RPC), a much cheaper source of dietary protein, has become commercially available for use in the feed industry. Importantly, dietary supplementation with RPC can increase feed intake by early‐weaned pigs. The objective of this study was to determine whether RPC can replace milk protein in the diet for early‐weaned pigs. RESULTS: Neither average daily gain (ADG), average daily feed intake (ADFI), nor the feed/gain ratio differed among the treatment groups in weeks 1 and 2. In week 3, the addition of 5 or 10% RPC to diets increased (P < 0.05) ADFI and ADG of pigs compared to those in the control group fed a 60% dried whey diet. During the entire 21‐day trial, ADFI and ADG were greater (P < 0.05) in pigs fed the 5 and 10% RCP diets than in pigs fed the 60% dried whey and 15% RCP diets. There were no differences in the serum concentrations of growth hormone on days 14 and 21, serum concentrations of insulin growth factor‐I (IGF‐I) on day 14, or IGF‐I gene expression in liver and skeletal muscle on days 14 and 21 among the dietary treatments. Serum concentrations of IGF‐I in pigs fed the 5, 10 and 15% RPC diets were greater than those in pigs fed the 60% dried whey diet. CONCLUSION: Findings indicate that up to 10% RPC can be used to replace dried whey in the diet for 7‐ to 21‐day‐old weaned piglets and can improve their growth performance. Copyright © 2008 Society of Chemical Industry     

Enhanced contractile force generation by artificial skeletal muscle tissues using IGF-I gene-engineered myoblast cells

The aim of this study was to investigate whether insulin-like growth factor (IGF)-I gene delivery to myoblast cells promotes the contractile force generated by hydrogel-based tissue-engineered skeletal muscles in vitro. Two retroviral vectors allowing doxycycline (Dox)-inducible expression of the IGF-I gene were transduced into mouse myoblast C2C12 cells to evaluate the effects of IGF-I gene expression on these cells. IGF-I gene expression stimulated the proliferation of C2C12 cells, and a significant increase in the growth rate was observed for IGF-I-transduced C2C12 cells with Dox addition, designated C2C12/IGF (Dox+) cells. Quantitative morphometric analyses showed that the myotubes induced from C2C12/IGF (Dox+) cells had a larger area and a greater width than control myotubes induced from normal C2C12 cells. Artificial skeletal muscle tissues were prepared from the respective cells using hydrogels composed of type I collagen and Matrigel. Western blot analyses revealed that the C2C12/IGF (Dox+) tissue constructs showed activation of a skeletal muscle hypertrophy marker (Akt) and enhanced expression of muscle-specific markers (myogenin, myosin heavy chain and tropomyosin). Moreover, the creatine kinase activity was increased in the C2C12/IGF (Dox+) tissue constructs. The C2C12/IGF (Dox+) tissue constructs contracted in response to electrical pulses, and generated a significantly higher physical force than the control C2C12 tissue constructs. These findings indicate that IGF-I gene transfer has the potential to yield functional skeletal muscle substitutes that are capable of in vivo restoration of the load-bearing function of injured muscle or acting as in vitro electrically-controlled bio-actuators.     

Fucosylated Chondroitin Sulfate from Sea Cucumber Improves Insulin Sensitivity via Activation of PI3K/PKB Pathway

This study was to investigate the effects of fucosylated chondroitin sulfate (CHS) from sea cucumber on insulin sensitivity in skeletal muscle of type 2 diabetic mice induced by a high‐fat high‐sucrose diet (HFSD). CHS supplementation for 19 wk significantly improved insulin sensitivity by 20%, and reduced blood glucose and insulin levels. Western blotting assay showed that CHS significantly increased insulin‐stimulated glucose transporter 4 (GLUT4) translocation to 1.7‐fold, phosphorylation of phosphoinositide 3‐kinase (PI3K) at p85 to 5.0‐fold, protein kinase B (PKB) at Ser473 to 1.5‐fold, and Thr308 to 1.6‐fold in skeletal muscle. However, PI3K, PKB, and GLUT4 total proteins expression were unchangeable. In addition, qRT‐PCR analysis proved that the insulin signaling was activated by CHS treatment, showing the increased mRNA expressions of glucose uptake‐related key genes. It indicated that CHS improved insulin sensitivity by activation of PI3K/PKB signaling in skeletal muscle of type 2 diabetic mice. Identification of potential mechanism by which CHS increased insulin sensitivity might provide a new functional food or pharmaceutical application of sea cucumber.     

Resveratrol Improves Muscle Atrophy by Modulating Mitochondrial Quality Control in STZ‐Induced Diabetic Mice

1 Scope

In this study, we aim to determine the effects of resveratrol (RSV) on muscle atrophy in streptozocin‐induced diabetic mice and to explore mitochondrial quality control (MQC) as a possible mechanism.

2 Methods and results

The experimental mice were fed either a control diet or an identical diet containing 0.04% RSV for 8 weeks. Examinations were subsequently carried out, including the effects of RSV on muscle atrophy and muscle function, as well as on the signaling pathways related to protein degradation and MQC processes. The results show that RSV supplementation improves muscle atrophy and muscle function, attenuates the increase in ubiquitin and muscle RING‐finger protein‐1 (MuRF‐1), and simultaneously attenuates LC3‐II and cleaved caspase‐3 in the skeletal muscle of diabetic mice. Moreover, RSV treatment of diabetic mice results in an increase in mitochondrial biogenesis and inhibition of the activation of mitophagy in skeletal muscle. RSV also protects skeletal muscle against excess mitochondrial fusion and fission in the diabetic mice.

3 Conclusion

The results suggest that RSV ameliorates diabetes‐induced skeletal muscle atrophy by modulating MQC.     

Influence of energy balance on the somatotrophic axis and matrix metalloproteinase expression in the endometrium of the postpartum dairy cow

Postpartum dairy cows enter a period of negative energy balance (NEB) associated with low circulating IGF1, during which the uterus must undergo extensive repair following calving. This study investigated the effects of NEB on expression of IGF family members and related genes in the involuting uterus. Cows were allocated to two treatments using differential feeding and milking regimes to produce mild NEB or severe NEB (SNEB). Uterine endometrial samples collected 2 weeks post partum were analysed by quantitative PCR. The expression of IGF-binding protein 4 (IGFBP4) mRNA increased in the endometrium of SNEB cows, with trends towards increased IGFBP1 and reduced IGFBP6 expression. There were no significant differences between treatments in mRNA expression of IGF1, IGF2 or of any hormone receptor studied, but significant correlations across all cows in the expression levels of groups of receptors suggested common regulatory mechanisms: type 1 IGF receptor (IGF1R), IGF2R and insulin receptor (INSR); GHR with ESR1; and ESR2 with NR3C1. The expression of IGF1R and INSR also positively correlated with the circulating urea concentration. Matrix metalloproteinases (MMPs) are important in tissue remodelling and can affect IGF signalling via interaction with IGFBPs. The expression levels of MMP1, MMP3, MMP9 and MMP13 mRNAs all showed major upregulation in the endometrium of cows in SNEB and all except MMP9 were highly correlated with expression of IGFBP4. Alpha(2)-HS-glycoprotein (AHSG) and PDK4, two genes implicated in insulin resistance, were also highly expressed in SNEB. These results suggest that cows in SNEB experience alterations to the IGF and insulin signalling pathways in the postpartum endometrium. This may affect the rate of tissue repair with a possible negative impact on subsequent fertility.     

Effect of L-carnitine infusion and feed restriction on carnitine status in lactating Holstein cows

Previously we determined that abomasal infusion of l-carnitine increased in vitro hepatic fatty acid oxidation, decreased liver lipid accumulation, and supported higher fat-corrected milk yield in feed-restricted lactating cows. The objectives of this study were to examine the effects of supplemental l-carni-tine and amount of feed intake on free carnitine and carnitine ester concentrations in liver, muscle, milk, and plasma of lactating dairy cows. Eight lactating Holstein cows (132 ± 36 d in milk) were used in a replicated 4 × 4 Latin square design with 14-d periods to test factorial combinations of water or l-carnitine infusion (20 g/d; d 5 to 14) and ad libitum or restricted (50% of previous 5-d intake; d 10 to 14) dry matter intake. Plasma was obtained 3 times daily on d 4, 8, and 12; milk samples were collected on d 8, 9, 13, and 14. Liver and muscle were biopsied on d 14 of each period. Free carnitine, short-chain acylcarnitine, and long-chain acylcarnitine concentrations were determined using a radioenzymatic assay coupled with ion exchange chromatography. Abomasal l-carnitine infusion increased total carnitine in plasma on d 8 and d 12. All liver carnitine fractions were increased by carnitine infusion. Feed restriction elevated concentrations of free carnitine, long-chain acylcarnitine, and total carnitine in liver tissue from carnitine-infused cows but not in those infused with water. In muscle, acid-soluble carnitine, long-chain acylcarnitine, and total carnitine concentrations were increased by carnitine infusion and feed restriction without significant interaction. Feed restriction increased free carnitine concentrations in muscle from water-infused cows but not in carnitine-infused cows. Carnitine infusion increased the concentration of each milk carnitine fraction as well as milk carnitine output on d 8 to 9. On d 13 to 14, all carnitine fractions except short-chain acylcarnitine were increased in milk from water-infused, feed-restricted cows, whereas all fractions were increased in carnitine-infused, feed-restricted cows. Carnitine infusion increased total carnitine in plasma, liver, muscle, and milk during feed restriction, whereas feed restriction alone increased carnitine concentrations in muscle and milk but not in liver. Liver carnitine concentrations might limit hepatic fatty acid oxidation capacity in dairy cows during the periparturient period; therefore, supplemental l-carnitine might decrease liver lipid accumulation in periparturient cows.     

青稞提取物干预对老年小鼠骨骼肌萎缩的影响

衰老过程中氧化应激的增加与骨骼肌质量与功能的显著降低存在密切关联。青稞富含多酚、类黄酮等生物活性成分,具有较强的抗氧化活性。研究表明,膳食中的天然抗氧化成分可以有效缓解骨骼肌增龄性萎缩。为探讨青稞水提物和醇提物是否对老年小鼠骨骼肌萎缩产生影响,测定了12周青稞提取物干预对老年小鼠骨骼肌年龄相关生理变化的影响,并利用反转录聚合酶链反应和免疫蛋白印记测定了肌萎缩标志基因、氧化应激相关基因的表达。实验结果表明:与衰老的对照小鼠相比,两种青稞提取物干预均有效抑制了衰老小鼠骨骼肌肌肉质量的减少,并增强了衰老小鼠的运动能力；青稞提取物在mRNA和蛋白水平均显著降低了肌肉环状指基因1和肌肉萎缩盒F基因的表达；青稞提取物激活了SIRT3蛋白的表达,并显著抑制了衰老小鼠骨骼肌中的氧化应激。研究结果表明,青稞提取物可以通过激活SIRT3信号的表达有效抑制衰老诱导的氧化应激,从而缓解衰老导致的骨骼肌萎缩。     

Dietary L-carnitine affects periparturient nutrient metabolism and lactation in multiparous cows

The objectives of this study were to determine the effects of dietary l-carnitine supplementation on liver lipid accumulation, hepatic nutrient metabolism, and lactation in multiparous cows during the periparturient period. Cows were assigned to treatments at d −25 relative to expected calving date and remained on the experiment until 56 d in milk. Treatments were 4 amounts of supplemental dietary carnitine: control (0 g/d of l-carnitine; n = 14); low carnitine (LC, 6 g/d; n = 11); medium carnitine (MC, 50 g/d; n = 12); and high carnitine (HC, 100 g/d; n = 12). Carnitine was supplied by mixing a feed-grade carnitine supplement with 113.5 g of ground corn and 113.5 g of dried molasses, which was then fed twice daily as a topdress to achieve desired daily carnitine intakes. Carnitine supplementation began on d −14 relative to expected calving and continued until 21 d in milk. Liver and muscle carnitine concentrations were markedly increased by MC and HC treatments. Milk carnitine concentrations were elevated by all amounts of carnitine supplementation, but were greater for MC and HC than for LC during wk 2 of lactation. Dry matter intake and milk yield were decreased by the HC treatment. The MC and HC treatments increased milk fat concentration, although milk fat yield was unaffected. All carnitine treatments decreased liver total lipid and triacylglycerol accumulation on d 10 after calving. In addition, carnitine-supplemented cows had higher liver glycogen during early lactation. In general, carnitine supplementation increased in vitro palmitate β-oxidation by liver slices, with MC and HC treatments affecting in vitro palmitate metabolism more potently than did LC. In vitro conversion of Ala to glucose by liver slices was increased by carnitine supplementation independent of dose. The concentration of nonesterified fatty acids in serum was not affected by carnitine. As a result of greater hepatic fatty acid β-oxidation, plasma β-hydroxybutyric acid was higher for the MC and HC treatments. Serum insulin was greater for all carnitine treatments, although plasma glucose was unaffected. Plasma urea N was lower and plasma total protein was higher for the MC and HC treatments. By decreasing liver lipid accumulation and stimulating hepatic glucose output, carnitine supplementation might improve glucose status and diminish the risk of developing metabolic disorders during early lactation.     

Effect of orally administered phenethyl isothiocyanate on hepatic gene expression in rats

Scope: Phenethyl isothiocyanate (PEITC) is a constituent of cruciferous vegetables that has demonstrated cancer preventive activity in a number of cancer models including lung, prostate, and breast cancer. Our objective was to examine the effects of the oral administration of PEITC for 7 days on the hepatic expression of genes important in drug metabolism and toxicity in Sprague Dawley rats. The liver is the major site for the metabolism of various xenobiotics and carcinogens, and determining the effects of PEITC on the gene expression of hepatic enzymes may provide insight into mechanisms underlying the cancer preventive activity of PEITC. Methods and results: Using a microarray containing 282 genes, we observed that PEITC significantly up‐regulated UDP‐glucuronosyltransferase UGT1A6 and strongly down‐regulated nicotinamide N‐methyltransferase (NNMT). We also confirmed the down‐regulation of NNMT by real‐time quantitative RT‐PCR. Other genes that were significantly up‐regulated were the drug metabolizing enzyme cyp2b15, the anti‐apoptotic gene bcl2l2, and the stress regulators Gadd45b, Dnajb9, Dnajb5 and Hspb1. Conclusion: Our results indicate new targets that may be important in the mechanisms of the anticancer effects of PEITC. Of particular significance was the down‐regulation of NNMT which may represent a new target for the treatment of a variety of cancers.     

Temporal changes in glycogenolytic enzyme mRNAs during myogenesis of primary porcine satellite cells

The objective was to study the regulation of glycogenolytic enzyme mRNAs in porcine satellite cells during proliferation and differentiation. Beyond 80% confluence, cells were grown in absence or presence of 1μM insulin. The observed increases in abundance of mRNA for glycogenin, glycogen synthase, phosphorylase kinase, phosphorylase and glycogen debranching enzyme, and no alterations of the transporter molecule GLUT4, clearly indicate that glycogenolytic enzymes of potential importance to meat quality development are regulated at the gene level during myogenesis, and are heavily involved in muscle cell and muscle fibre development. The genes, however, are not influenced by insulin, and the lack of response to insulin of expression of gene-encoding enzymes involved in the formation and degradation of glycogen may question the applicability of porcine cell culture systems, like the one applied, as a model to study the regulation and regulatory mechanism of energy metabolism in muscles.     

Voluntary exercise and green tea enhance the expression of genes related to energy utilization and attenuate metabolic syndrome in high fat fed mice

Obesity and metabolic syndrome are growing public health problems. We investigated the effects of decaffeinated green tea extract (GTE) and voluntary running exercise (Ex) alone or in combination against obesity and metabolic syndrome in high fat (HF) fed C57BL/6J mice. After 16 wk, GTE + Ex treatment reduced final body mass (27.1% decrease) and total visceral fat mass (36.6% decrease) compared to HF‐fed mice. GTE + Ex reduced fasting blood glucose (17% decrease), plasma insulin (65% decrease), and insulin resistance (65% decrease) compared to HF‐fed mice. GTE or Ex alone had less significant effects. In the skeletal muscle, the combination of Ex and GTE increased the expression of peroxisome proliferator‐activated receptor‐γ coactivator‐1α (Ppargc1a), mitochondrial NADH dehydrogenase 5 (mt‐Nd5), mitochondrial cytochrome b (mt‐Cytb), and mitochondrial cytochrome c oxidase III (mt‐Co3). An increase in hepatic expression of peroxisome proliferator‐activated receptor‐α (Ppara) and liver carnitine palmitoyl transferase‐1α (Cpt1a) and a decrease in hepatic expression of stearoyl‐CoA desaturase 1 (Scd1) mRNA was observed in GTE + Ex mice. GTE + Ex was more effective than either treatment alone in reducing diet‐induced obesity. These effects are due in part to modulation of genes related to energy metabolism and de novo lipogenesis.     

花椒麻素对SD雄性大鼠骨骼肌蛋白质代谢的影响及机理

为进一步了解花椒麻素对机体蛋白质代谢的影响及调控机理,将40 只健康SD雄性大鼠按体质量随机分为 空白对照组、花椒麻素的大豆油溶液高剂量灌胃组（8 mg/（kg·d））、中剂量灌胃组（4 mg/（kg·d））和低剂 量灌胃组（2 mg/（kg·d））,各组10 只,饲养28 d。实验结束后,测定血清中血尿素氮、总蛋白、胰岛素样生长 因子（insulin-like growth factorⅠ,IGF-Ⅰ）、氨基酸和骨骼肌中氨基酸水平及变化;实时荧光定量聚合酶链式反 应检测肝脏中IGF-Ⅰ、胰岛素样生长因子受体-Ⅰ（insulin growth factorⅠreceptor,IGF-ⅠR）的mRNA转录水平, 骨骼肌中IGF-Ⅰ、IGF-ⅠR、肌细胞生成素（myogenin,MyoG）、肌肉生长抑制素（myostatin,MSTN）、钙蛋白 酶-1（μ-calpain,CAPN-1）、钙蛋白酶抑制蛋白（calpastatin,CAST）的mRNA转录水平。结果表明：与空白对照 组相比,花椒麻素可显著提高大鼠相对骨骼肌的质量,降低腹脂率,对其他脏器的相对质量没有显著影响;显著降 低大鼠血尿氮水平;显著升高大鼠血清和骨骼肌总蛋白质水平。此外,花椒麻素可显著上调大鼠骨骼肌中IGF-Ⅰ、 MyoG、CAPN-1和CAST mRNA相对表达量,显著下调MSTN mRNA相对表达量。花椒麻素提高大鼠骨骼肌相对质 量的主要机制是通过上调MyoG mRNA表达量的同时下调抑制基因MSTN mRNA相对表达量,促进骨骼肌的生长; 另外花椒麻素可由CAPN-1/CAST系统增强骨骼肌蛋白质的更新,并促进蛋白质的沉积。     

Lipid metabolic effect of Korean red ginseng extract in mice fed on a high-fat diet

BACKGROUND: Ginseng saponin and ginsenosides exert anti‐obesity effects via the modulation of physiological lipid metabolism in vivo or intracellular signalling in cell culture systems. However, the complicated relationship between the anti‐obesity effects of ginseng and gene expression has yet to be defined under in vivo conditions. Therefore, we evaluated the relationship between the anti‐obesity effects of Korean red ginseng extract (KRGE) and hepatic gene expression profiles in mice fed long‐term on a high‐fat diet (HFD) in this study. RESULTS: KRGE reduces the levels of cholesterol, low‐density lipoprotein‐cholesterol (LDL‐C), serum triglycerides, and atherogenic indices. Levels of leptin, adiponectin and insulin, which regulate glucose and lipid metabolism, were impaired profoundly by HFD. However, KRGE treatment brought these levels back to normal. KRGE was found to down‐regulate genes associated with lipid metabolism or cholesterol metabolism (Lipa, Cyp7a1, Il1rn, Acot2, Mogat1, Osbpl3, Asah3l, Insig1, Anxa2, Vldlr, Hmgcs1, Sytl4, Plscr4, Pla2g4e, Slc27a3, Enpp6), all of which were up‐regulated by HFD. CONCLUSION: KRGE regulated the expression of genes associated with abnormal physiology via HFD. Leptin, insulin, and adiponectin, which carry out critical functions in energy and lipid metabolism, were shown to be modulated by KRGE. These results show that KRGE is effective in preventing obesity. Copyright © 2011 Society of Chemical Industry