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Creatine nitrate supplementation strengthens energy status and delays glycolysis of broiler muscle via inhibition of LKB1/AMPK pathway.

ABSTRACT: This study aimed to evaluate the effects of dietary creatine nitrate (CrN) on growth performance, meat quality, energy status, glycolysis, and related gene expression of liver kinase B1/AMP-activated protein kinase (LKB1/AMPK) pathway in Pectoralis major (PM) muscle of broilers. A total of 240 male Arbor Acres broilers (28-day-old) were randomly allocated to one of 5 dietary treatments: the basal diet (control group), and the basal diets supplemented with 600 mg/kg guanidinoacetic acid (GAA), 300, 600, or 900 mg/kg CrN (identified as GAA₆₀₀, CrN₃₀₀, CrN₆₀₀, or CrN₉₀₀, respectively). We found that dietary GAA and CrN supplementation for 14 d from d 28 to 42 did not affect broiler growth performance, carcass traits, and textural characteristics of breast muscle. GAA₆₀₀, CrN₆₀₀, and CrN₉₀₀ treatments increased pH_24h and decreased drip loss of PM muscle compared with the control (P < 0.05). The PM muscles of CrN₆₀₀ and CrN₉₀₀ groups showed higher glycogen concentration and lower lactic acid concentration accompanied by lower activities of phosphofructokinase (PFK), pyruvate kinase (PK), and lactate dehydrogenase (LDH) (P < 0.05). Simultaneously, GAA₆₀₀ and all CrN treatments increased concentration of muscle creatine, phosphocreatine (PCr) and ATP, and decreased AMP concentration and AMP/ATP ratio (P < 0.05). Meanwhile, the concentrations of muscle creatine, PCr, and ATP were increased linearly, while muscle AMP concentration and AMP/ATP ratio were decreased linearly and quadratic as the dose of CrN increased (P < 0.05). GAA₆₀₀, CrN₆₀₀, and CrN₉₀₀ treatments upregulated mRNA expression of CreaT in PM muscle, and CrN₆₀₀ and CrN₉₀₀ treatments downregulated GAMT expression in liver and PM muscle compared with the control or GAA₆₀₀ groups (P < 0.05). The mRNA expression of muscle LKB1, AMPKα1, and AMPKα2 was downregulated linearly in response to the increasing CrN level (P < 0.05). Overall, CrN showed better efficacy on strengthening muscle energy status and improve meat quality than GAA at the some dose. These results indicate that CrN may be a potential replacement for GAA as a new creatine supplement.

SUBMITTER: Duan BB

PROVIDER: S-EPMC8749301 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:This study investigated the attenuating effects of dietary creatine nitrate (CrN), a novel form of creatine, on energy expenditure and rapid glycolysis in pectoralis major (PM) muscle of broiler induced by preslaughter transport. A total of 288 Arbor Acres broilers (28 day old) were randomly assigned into five dietary treatments, including a basal diet or the basal diet supplemented with 600 mg/kg guanidinoacetic acid (GAA), 300, 600, or 900 mg/kg CrN for 14 d, respectively. On the day of transportation, the broilers from basal diet group were divided into two equal groups: one group was transported for 0.5 h (Control group) and the other group was transported for 3 h (T3h group). Meanwhile, the birds from GAA and CrN supplementation groups were transported for 3 h (identified as GAA600, CrN300, CrN600, and CrN900 group, respectively). The results demonstrated that dietary supplementation of GAA or CrN from 28 to 42 d of age did not significantly affect the growth performance, carcass traits, and textural characteristics (P > 0.05) in PM muscle of transported broilers. Compared with T3h group, GAA600, CrN600, and CrN900 groups increased the pH45min (P < 0.01), and CrN600, CrN900 groups decreased the cooking loss (P < 0.05) of PM muscle. Meanwhile, the muscle of GAA600, CrN600, and CrN900 groups showed a higher glycogen content (P < 0.01) and a lower lactic acid content (P < 0.01). GAA600 and all CrN treatments enhanced muscle Cr content and reduced AMP/ATP ratio (P < 0.01). In addition, GAA600 and all CrN treatments downregulated the relative mRNA expression levels of LKB1 and AMPKα2 (P < 0.001) and the protein expression of p-AMPKαThr172 compared with the T3h group (P < 0.01). All CrN treatments showed lower protein expression levels of LKB1 and p-LKB1Thr189 than those of the T3h group (P < 0.05). In summary, dietary supplementation with GAA and CrN enhanced the content of muscle creatine, and inhibited transport-induced activation of LKB1/AMPK pathway, which is beneficial for delaying rapid muscle glycolysis and improving meat quality.

Project description:Oxidative stress is common in the whole process of broiler production, and breast muscle is one of the target organs most vulnerable to oxidative attack. When broilers are subjected to oxidative stress, the regulation of adenosine 5-monophosphate activated protein kinase (AMPK) is a critical path to maintain the dynamic balance of intracellular energy. However, whether calcium/calmodulin-dependent protein kinase (CaMKK) and liver kinase B1 (LKB1) are involved in the regulation of AMPK activation in broiler breast muscle under oxidative stress has not been elucidated. In this study, a total of 144 one-day-old male Ross 308 chicks were selected, with an average body weight of 43.44 ± 0.04 g. The broilers were divided into 3 groups with 6 replicates of 8 broilers each (control group, intraperitoneal injection of physiological saline group, and intraperitoneal injection of hydrogen peroxide [H2O2] group), the injection time was selected on the 16th and 37th day of the experimental period, the injection volumes were 1.0 mL/kg broiler body weight. The results of this experiment showed that H2O2 exposure reduced the average daily gain (ADG) and increased the feed to gain ratio (F/G), the level of corticosterone (CORT) and the activity of lactate dehydrogenase (LDH) in serum were increased after H2O2 exposure. H2O2 exposure also increased the contents of reactive oxygen species (ROS) and protein carbonyl, but decreased the activities of catalase (CAT), total antioxidant capacity (T-AOC), total superoxide dismutase (T-SOD) and glutathione peroxidase (GSH-Px) in breast muscle. After H2O2 exposure, the activity of pyruvate dehydrogenase (PDH) was decreased, the content of glycogen was reduced, and the contents of adenosine monophosphate (AMP) and lactate were increased in breast muscle. In addition, H2O2 exposure increased the content of Ca2+, upregulated the protein expression levels of CaMKK1 and p-AMPK, and increased the activities of hexokinase (HK) and LDH in breast muscle. These findings suggested that the activation of CaMKK/LKB1/AMPK signaling pathway would be associated with the accelerated glycolysis of broiler breast muscle under oxidative stress.

Project description:In a double-blind, crossover, randomized and placebo-controlled trial; 28 men and women ingested a placebo (PLA), 3 g of creatine nitrate (CNL), and 6 g of creatine nitrate (CNH) for 6 days. Participants repeated the experiment with the alternate supplements after a 7-day washout. Hemodynamic responses to a postural challenge, fasting blood samples, and bench press, leg press, and cycling time trial performance and recovery were assessed. Data were analyzed by univariate, multivariate, and repeated measures general linear models (GLM). No significant differences were found among treatments for hemodynamic responses, clinical blood markers or self-reported side effects. After 5 days of supplementation, one repetition maximum (1RM) bench press improved significantly for CNH (mean change, 95% CI; 6.1 [3.5, 8.7] kg) but not PLA (0.7 [-1.6, 3.0] kg or CNL (2.0 [-0.9, 4.9] kg, CNH, p = 0.01). CNH participants also tended to experience an attenuated loss in 1RM strength during the recovery performance tests following supplementation on day 5 (PLA: -9.3 [-13.5, -5.0], CNL: -9.3 [-13.5, -5.1], CNH: -3.9 [-6.6, -1.2] kg, p = 0.07). After 5 days, pre-supplementation 1RM leg press values increased significantly, only with CNH (24.7 [8.8, 40.6] kg, but not PLA (13.9 [-15.7, 43.5] or CNL (14.6 [-0.5, 29.7]). Further, post-supplementation 1RM leg press recovery did not decrease significantly for CNH (-13.3 [-31.9, 5.3], but did for PLA (-30.5 [-53.4, -7.7] and CNL (-29.0 [-49.5, -8.4]). CNL treatment promoted an increase in bench press repetitions at 70% of 1RM during recovery on day 5 (PLA: 0.4 [-0.8, 1.6], CNL: 0.9 [0.35, 1.5], CNH: 0.5 [-0.2, 0.3], p = 0.56), greater leg press endurance prior to supplementation on day 5 (PLA: -0.2 [-1.6, 1.2], CNL: 0.9 [0.2, 1.6], CNH: 0.2 [-0.5, 0.9], p = 0.25) and greater leg press endurance during recovery on day 5 (PLA: -0.03 [-1.2, 1.1], CNL: 1.1 [0.3, 1.9], CNH: 0.4 [-0.4, 1.2], p = 0.23). Cycling time trial performance (4 km) was not affected. Results indicate that creatine nitrate supplementation, up to a 6 g dose, for 6 days, appears to be safe and provide some ergogenic benefit.

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Creatine nitrate supplementation strengthens energy status and delays glycolysis of broiler muscle via inhibition of LKB1/AMPK pathway.

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