Increased Carnosine Content Research Articles

RNA Profiles of the Korat Chicken Breast Muscle with Increased Carnosine Content Produced through Dietary Supplementation with β-Alanine or L-Histidine.

Simple SummaryCarnosine is a bioactive food component with several potential health benefits for humans due to its physiological functions. Dietary supplementation with β-alanine or L-histidine can increase the carnosine content of skeletal muscles in chickens. Dietary supplementation with β-alanine or L-histidine has produced a slow-growing chicken variety with high carnosine content in the breast meat; however, the supplementation with L-histidine alone softens the meat toughness, which may affect consumers’ willingness to buy the meat. Gene expression is a key factor that influences meat quality. Understanding the molecular mechanisms that affect carnosine content and meat toughness would allow the production of more value-added slow-growing chickens. We compared global gene expression in chicken breast muscles with differing carnosine contents and meat toughness produced through dietary supplementation with β-alanine or L-histidine. We identified differentially expressed genes involved in regulating myosin, collagen, intramuscular fat, and calpain—factors that may affect meat tenderness. Pathway enrichment analysis indicated that the insulin-related and adipocytokine signaling pathways were altered by dietary supplementation with β-alanine or L-histidine. These data will be useful for future studies on carnosine content and meat toughness in slow-growing chickens.Korat chicken (KRC) is a slow-growing chicken bred in Thailand, whose meat exhibits a unique toughness. A previous study produced KRC breast meat containing high carnosine content through dietary supplementation with β-alanine or L-histidine; however, the KRC that were fed an L-histidine-supplemented diet produced meat that was significantly more tender. Herein, we performed RNA-Seq to identify candidate genes involved in the regulation of carnosine content and meat toughness. Total RNA was isolated from five female KRC breast muscles in each treatment group that KRC fed diets without supplementation, supplemented with β-alanine or L-histidine. Compared to the non-supplemented group, we identified 118 and 198 differentially expressed genes (DEGs) in the β-alanine or L-histidine supplementation groups, respectively. Genes potentially related to meat tenderness—i.e., those regulating myosin, collagen, intramuscular fat, and calpain—were upregulated (LOC107051274, ACSBG1, and CAPNS2) and downregulated (MYO7B, MYBPH, SERPINH1, and PGAM1). However, carnosine synthase gene was not identified. Functional enrichment analysis identified pathways affected by dietary supplementation, including the insulin signaling pathway (β-alanine supplementation) and the insulin resistance and adipocytokine signaling pathways (L-histidine supplementation). The FoxO signaling pathway was identified as a regulatory network for both supplementation groups. The identified genes can be used as molecular markers of meat tenderness in slow-growing chickens.

Antioxidative Characteristics of Chicken Breast Meat and Blood after Diet Supplementation with Carnosine, L-histidine, and β-alanine.

The objective of the study was to test the effect of diets supplemented with β-alanine, L-histidine, and carnosine on the histidine dipeptide content and the antioxidative status of chicken breast muscles and blood. One-day-old Hubbard Flex male chickens were assigned to five treatments: control diet (C) and control diet supplemented with 0.18% L-histidine (ExpH), 0.3% β-alanine (ExpA), a mix of L-histidine\\β-alanine (ExpH+A), and 0.27% carnosine (ExpCar). After 28 days, chicken breast muscles and blood samples were analyzed for the antioxidant enzyme activity (catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD)), carnosine and anserine content, amino acid profile, and anti-radical activity (ABTS, DPPH, ferric reducing antioxidant power (FRAP)). The results of the study showed that carnosine supplementation effectively increased body weight and breast muscle share in chicken carcasses. Carnosine and L-histidine supplementation with or without β-alanine increased carnosine content in chicken breast muscles up to 20% (p = 0.003), but the boost seems to be too low to affect the potential antioxidant capacity and amino acid content. The β-alanine-enriched diet lowered dipeptide concentration in chicken blood serum (p = 0.002) and activated catalase in chicken breast muscles in relation to the control group (p = 0.003). It can be concluded that histidine or dipeptide supplementation of chicken diets differently affected the total antioxidant potential: in breast muscles, it increased dipeptide content, while in blood cell sediment (rich in erythrocytes), increased SOD and GPx activities were observed.

Effect of β-alanine supplementation on carnosine and histidine content in the hippocampus of 14-month-old rats.

Carnosine and histidine content in the hippocampus of 14-month-old male rats was examined following 30 days of β-alanine supplementation. All animals were provided identical diets; however, 100 mg of β-alanine was mixed with glucomannan (80:20 blend) in the water of β-alanine supplemented animals. Hippocampal carnosine content was significantly greater (p = 0.005) for β-alanine compared with control, while no differences (p = 0.438) were noted in histidine content between groups. Results provide initial evidence that β-alanine supplementation increases carnosine content in the hippocampus of middle-aged rats, without compromising histidine content. β-Alanine supplementation increases hippocampal carnosine content without compromising histidine content in middle-aged rats.

Exercise and β-alanine supplementation on carnosine-acrolein adduct in skeletal muscle

Previous studies have demonstrated that exercise results in reactive aldehyde production and that β-alanine supplementation increases carnosine content in skeletal muscle. However, little is known about the influence exercise and β-alanine supplementation have on the formation of carnosine-aldehydes. The goal of the present study was to monitor the formation of carnosine-aldehyde adducts, following high-intensity intermittent exercise, before and after β-alanine supplementation. Vastus lateralis biopsy samples were taken from 14 cyclists, before and after a 28 day β-alanine supplementation, following 4 bouts of a 30 s all-out cycling test, and carnosine and CAR-aldehyde adducts [carnosine-acrolein, CAR-ACR (m/z 303), carnosine-4-hydroxy-2-hexenal, CAR-HHE (m/z 341) and carnosine-4-hydroxy-2-nonenal, CAR-HNE (m/z 383)] were quantified by HPLC-MS/MS. β-alanine supplementation increased muscle carnosine content by ~50% (p = 0.0001 vs. Pre-Supplementation). Interestingly, there was a significant increase in post-exercise CAR-ACR content following β-alanine supplementation (p < 0.001 vs. post-exercise before supplementation), whereas neither exercise alone nor supplementation alone increased CAR-ACR formation. These results suggest that carnosine functions as an acrolein-scavenger in skeletal muscle. Such a role would be relevant to the detoxification of this aldehyde formed during exercise, and appears to be enhanced by β-alanine supplementation. These novel findings not only have the potential of directly benefiting athletes who engage in intensive training regimens, but will also allow researchers to explore the role of muscle carnosine in detoxifying reactive aldehydes in diseases characterized by abnormal oxidative stress.

Influence of Dietary Histidine, Hybrid Line and Gender on Chicken Meat Quality and Carnosine Concentration

The aim of the investigation was to determine influence of L-histidine dietary supplementation (P1 0.0%, P2 0.1%, P3 0.2% and P4 0.3%), hybrid line (Cobb 500 and Hubbard Classic) and gender on meat quality and carnosine concentration in chicken breast and thigh muscles. The research was carried out on 184 one day old broiler chickens as 4x2x2 factorial design. Chicken live weight, carcass weight, dressing percentage, absolute and relative shares of main body parts in the carcass, pHi, pHu, colour, drip loss, cooking loss and share force of breast muscle, and carnosine concentration in breast and thigh muscles were determined. L-histidine dietary supplementation significantly affected live weight, carcass weight, weight of drumsticks and thighs, backs and wings, share of back and the CIE a* value (P<0.05), as well as the content of carnosine in breast muscle (P=0.003). Hybrid line influenced relative shares of main body parts in the carcasses, except for wings, and the CIE a* value of breast muscle (P < 0.001). Chicken gender affected live weight of chickens, carcass weight, as well as weight of main parts (P<0.001), except for breast. The results of the investigation showed that dietary L- histidine significantly increased carnosine content in broiler breast muscle. Cobb 500 female chickens of the P4 group had higher carnosine concentrations in breast muscle by 156.61 and male chickens by 150.49 µg/g of tissue than the control group. Cobb 500 broiler chickens deposited more carnosine in meat than Hubbard Classic broilers.

Muscle carnosine loading by beta-alanine supplementation is more pronounced in trained vs. untrained muscles

Carnosine occurs in high concentrations in human skeletal muscle and assists working capacity during high-intensity exercise. Chronic beta-alanine (BA) supplementation has consistently been shown to augment muscle carnosine concentration, but the effect of training on the carnosine loading efficiency is poorly understood. The aim of the present study was to compare muscle carnosine loading between trained and untrained arm and leg muscles. In a first study (n = 17), reliability of carnosine quantification by proton magnetic resonance spectroscopy ((1)H-MRS) was evaluated in deltoid and triceps brachii muscles. In a second study, participants (n = 35; 10 nonathletes, 10 cyclists, 10 swimmers, and 5 kayakers) were supplemented with 6.4 g/day of slow-release BA for 23 days. Carnosine content was evaluated in soleus, gastrocnemius medialis, and deltoid muscles by (1)H-MRS. All the results are reported as arbitrary units. In the nonathletes, BA supplementation increased carnosine content by 47% in the arm and 33% in the leg muscles (not significant). In kayakers, the increase was more pronounced in arm (deltoid) vs. leg (soleus + gastrocnemius) muscles (0.089 vs. 0.049), whereas the reverse pattern was observed in cyclists (0.065 vs. 0.084). Swimmers had significantly higher increase in carnosine in both deltoid (0.107 vs. 0.065) and gastrocnemius muscle (0.082 vs. 0.051) compared with nonathletes. We showed that 1) carnosine content can be reliably measured by (1)H-MRS in deltoid muscle, 2) carnosine loading is equally effective in arm vs. leg muscles of nonathletes, and 3) carnosine loading is more pronounced in trained vs. untrained muscles.

Effect of two β-alanine dosing protocols on muscle carnosine synthesis and washout

Carnosine (β-alanyl-L-histidine) is found in high concentrations in skeletal muscle and chronic β-alanine (BA) supplementation can increase carnosine content. This placebo-controlled, double-blind study compared two different 8-week BA dosing regimens on the time course of muscle carnosine loading and 8-week washout, leading to a BA dose-response study with serial muscle carnosine assessments throughout. Thirty-one young males were randomized into three BA dosing groups: (1) high-low: 3.2 g BA/day for 4 weeks, followed by 1.6 g BA/day for 4 weeks; (2) low-low: 1.6 g BA/day for 8 weeks; and (3) placebo. Muscle carnosine in tibialis-anterior (TA) and gastrocnemius (GA) muscles was measured by 1H-MRS at weeks 0, 2, 4, 8, 12 and 16. Flushing symptoms and blood clinical chemistry were trivial in all three groups and there were no muscle carnosine changes in the placebo group. During the first 4 weeks, the increase for high-low (TA 2.04 mmol/kgww, GA 1.75 mmol/kgww) was ~twofold greater than low-low (TA 1.12 mmol/kgww, GA 0.80 mmol/kgww). 1.6 g BA/day significantly increased muscle carnosine within 2 weeks and induced continual rises in already augmented muscle carnosine stores (week 4-8, high-low regime). The dose-response showed a carnosine increase of 2.01 mmol/kgww per 100 g of consumed BA, which was only dependent upon the total accumulated BA consumed (within a daily intake range of 1.6-3.2 g BA/day). Washout rates were gradual (0.18 mmol/kgww and 0.43 mmol/kgww/week; ~2%/week). In summary, the absolute increase in muscle carnosine is only dependent upon the total BA consumed and is not dependent upon baseline muscle carnosine, the muscle type, or the daily amount of supplemented BA.