Maternal betaine supplementation during gestation enhances gene expression in skeletal muscle of newborn piglets

Betaine has been widely used in animal and human nutrition to promote muscle growth and performance, yet it remains unknown whether maternal betaine supplementation during gestation affects the metabolic characteristics of neonatal skeletal muscles. In the present study, feeding sows with betaine-supplemented diets throughout gestation significantly up-regulated the expression of mtDNA-encoded OXPHOS genes including COX1, COX2 and ND5, in the muscle of newborn piglets, which was associated with enhanced mitochondrial COX enzyme activity. Concurrently, maternal betaine supplementation increased plasma betaine concentration and muscle expression of methyl transfer enzymes, BHMT and GNMT in offspring piglets. Nevertheless, Dnmt3a was down-regulated at the level of both mRNA and protein, which was associated with hypomethylated mtDNA D-loop region. These results suggest that maternal betaine supplementation during gestation enhances expression of mtDNA-encoded genes through D-loop DNA hypomethylation in skeletal muscle of newborn piglets.

Jia, Y., et al., Maternal betaine supplementation during gestation enhances expression of mtDNA-encoded genes through D-loop DNA hypomethylation in skeletal muscle of newborn piglets. J Agric Food Chem, 2015.

Betaine supplementation causes increase in carnitine metabolites in the muscle and liver of mice fed a high-fat diet as studied by nontargeted LC-MS metabolomics approach

SCOPE: Betaine (BET) reduces diet-induced liver lipid accumulation, and may relieve obesity-related metabolic disturbances. The aim of our study was to analyze metabolite alterations after supplementation of BET, polydextrose (PDX, a soluble dietary fiber), or their combination (BET PDX) via drinking water to C57BL/6J mice fed a high-fat (HF) diet.
METHODS AND RESULTS: BET supplementation increased BET levels in plasma, muscle, and liver (p < 0.05), and the nontargeted LC-MS metabolite profiling revealed an increase in several metabolites in the carnitine biosynthesis pathway after BET supplementation both in liver and muscle. These included carnitine and acetylcarnitine (1.4-fold, p < 0.05), propionylcarnitine and gamma-butyrobetaine (1.5-fold, p < 0.05), and several other short-chain acylcarnitines (p < 0.05) in muscle. These changes were slightly higher in the BET PDX group. Furthermore, BET reduced the HF diet induced accumulation of triglycerides in liver (p < 0.05). The supplementations did not attenuate the HF diet induced increase in body weight gain or the increase in adipose tissue mass. Instead, the combination of BET and PDX tended to increase adiposity. CONCLUSION: Our results suggest that increased availability of BET in different tissues, especially in muscle, after BET supplementation has an impact on carnitine metabolism, and this could further explain the link between BET and lipid metabolism.

Pekkinen, J., et al., Betaine supplementation causes increase in carnitine metabolites in the muscle and liver of mice fed a high-fat diet as studied by nontargeted LC-MS metabolomics approach. Mol Nutr Food Res, 2013

Betaine supplement enhances skeletal muscle differentiation in murine myoblasts via IGF-1 signaling activation

BACKGROUND: Betaine (BET) is a component of many foods, including spinach and wheat. It is an essential osmolyte and a source of methyl groups. Recent studies have hypothesized that BET might play a role in athletic performance. However, BET effects on skeletal muscle differentiation and hypertrophy are still poorly understood.
METHODS: We examined BET action on neo myotubes maturation and on differentiation process, using C2C12 murine myoblastic cells. We used RT2-PCR array, Western blot and immunofluorescence analysis to study the BET effects on morphological features of C2C12 and on signaling pathways involved in muscle differentiation and hypertrophy.
RESULTS: We performed a dose--response study, establishing that 10 mM BET was the dose able to stimulate morphological changes and hypertrophic process in neo myotubes. RT2-PCR array methodology was used to identify the expression profile of genes encoding proteins involved in IGF-1 pathway. A dose of 10 mM BET was found to promote IGF-1 receptor (IGF-1 R) expression. Western blot and immunofluorescence analysis, performed in neo myotubes, pointed out that 10 mM BET improved IGF-1 signaling, synthesis of Myosin Heavy Chain (MyHC) and neo myotubes length.In addition, we investigated BET role on myoblasts proliferation and differentiation. During proliferation, BET did not modify C2C12 proliferative rate, but promoted myogenic induction, enhancing MyoD protein content and cellular elongation. During differentiation, BET caused an increase of muscle-specific markers and IGF-1 R protein levels. CONCLUSIONS: Our findings provide the first evidence that BET could promote muscle fibers differentiation and increase myotubes size by IGF-1 pathway activation, suggesting that BET might represent a possible new drug/integrator strategy, not only in sport performance but also in clinical conditions characterized by muscle function impairment.

Senesi, P., et al., Betaine supplement enhances skeletal muscle differentiation in murine myoblasts via IGF-1 signaling activation. J Transl Med, 2013. 11(1): p. 174

Betaine supplementation enhances anabolic endocrine and Akt signaling in response to acute bouts of exercise

Our aim was to examine the effect of betaine supplementation on selected circulating hormonal measures and Akt muscle signaling proteins after an acute exercise session. Twelve trained men (age 19.7 +/- 1.23 years) underwent 2 weeks of supplementation with either betaine (B) (1.25 g BID) or placebo (P). Following a 2-week washout period, subjects underwent supplementation with the other treatment (B or P). Before and after each 2-week period, subjects performed an acute exercise session (AES). Circulating GH, IGF-1, cortisol, and insulin were measured. Vastus lateralis samples were analyzed for signaling proteins (Akt, p70 S6k, AMPK). B (vs. P) supplementation approached a significant increase in GH (mean +/- SD (Area under the curve, AUC), B: 40.72 +/- 6.14, P: 38.28 +/- 5.54, p = 0.060) and significantly increased IGF-1 (mean +/- SD (AUC), B: 106.19 +/- 13.45, P: 95.10 +/- 14.23, p = 0.010), but significantly decreased cortisol (mean +/- SD (AUC), B: 1,079.18 +/- 110.02, P: 1,228.53 +/- 130.32, p = 0.007). There was no difference in insulin (AUC). B increased resting Total muscle Akt (p = 0.003). B potentiated phosphorylation (relative to P) of Akt (Ser(473)) and p70 S6 k (Thr(389)) (p = 0.016 and p = 0.005, respectively). Phosphorylation of AMPK (Thr(172)) decreased during both treatments (both p = 0.001). Betaine (vs. placebo) supplementation enhanced both the anabolic endocrine profile and the corresponding anabolic signaling environment, suggesting increased protein synthesis.
Apicella, J.M., et al., Betaine supplementation enhances anabolic endocrine and Akt signaling in response to acute bouts of exercise. Eur J Appl Physiol, 2013. 113(3): p. 793-802

Betaine: a promising antioxidant agent for enhancement of broiler meat quality

1. Antioxidant and methyl donor effects of betaine in experimental animal models have recently been demonstrated. The present study was therefore designed to examine the antioxidant effects of betaine on the antioxidant status and meat quality of breast muscles in broilers.
2. Cobb broilers were randomly divided into Control, Methionine low, Methionine low plus betaine, and Betaine groups.
3. The activity of the main antioxidant enzyme (glutathione peroxidase) in the Betaine and the Methionine low plus betaine groups significantly increased compared to the Methionine low and Control groups. Catalase and superoxide dismutase activities were significantly higher in the Betaine group compared to the Methionine low group, and lipid peroxidation was significantly higher in the Control and the Methionine low groups.
4. The present study indicates that adding betaine (1 g/kg) to a diet deficient in methionine can significantly improve antioxidant defences and meat quality, decreasing lipid peroxidation in the breast muscles of broiler chickens.
Alirezaei, M., et al., Betaine: a promising antioxidant agent for enhancement of broiler meat quality. British Poultry Science, 2012. 53(5): p. 699-707

Effect of long-term betaine supplementation on chemical and physical characteristics of three muscles from the Alentejano pig

BACKGROUND: The use of betaine as a dietary supplement to reduce fat deposition and increase lean muscle mass in pigs is increasing. However, there is no available information on its effect on the physical and chemical (i.e. fatty acid (FA) profile) characteristics of pork. RESULTS: The effects of long-term betaine intake (1 g betaine kg(-1) diet for 20 weeks) on the chemical and physical characteristics of Musculus longissimus dorsi, M. biceps femoris and M. semimembranosus from the Alentejano pig were investigated. The contents of total protein, intramuscular lipids (neutral and polar), myoglobin and total collagen as well as the water-holding capacity, physical colour characteristics and FA profile of the three muscles were determined. With the exception of a significant increase in the intramuscular lipid content of M. longissimus dorsi and M. biceps femoris, betaine supplementation did not influence muscle chemical and colour characteristics or body fat deposition. CONCLUSION: These data suggest that long-term betaine supplementation selectively increases intramuscular lipid deposition (a trait related to meat quality) while not affecting other chemical (including the atheroprotective FA profile) and physical characteristics of the Alentejano pig muscles studied.
Martins, J.M., et al., Effect of long-term betaine supplementation on chemical and physical characteristics of three muscles from the Alentejano pig. J Sci Food Agric, 2012. 92(10): p. 2122-7

Muscle endurance and quality of repetitions

This study examined the efficacy of 15 days of betaine supplementation on muscle endurance, power performance and rate of fatigue in active college-aged men. Male subjects (24) were randomly assigned to one of two groups; the first consumed betaine daily (2.5 g/d), and the second consumed a placebo. Subjects were tested prior to the onset of supplementation (T1) and 7 (T2) and 14 days (T3) following supplementation. Subjects were tested over a 2-day period for:

- vertical jump power (VJP)
- bench press throw (BPT) power
- number of squat repetitions at 75% 1-RM (peak and mean power)
- number of bench press repetitions at 75% 1-RM (peak and mean power)
- two 30-sec Wingate anaerobic power tests (WAnT) (Day 2)

There were no differences in power assessment (VJP, BPT, WAnT) between the groups, or in the repetitions performed to exhaustion or in the number of repetitions performed at 90% of both peak and mean power between the groups in the bench press exercise.

However, the number of repetitions performed in the squat exercise for BET was significantly greater (p < 0.05) than that seen for PL at T2. The number of repetitions performed at 90% or greater of peak power in the squat exercise was significantly greater for BET at both T2 and T3 than PL. It was concluded that either 1 or 2 weeks of betaine supplementation in active, college males improved muscle endurance of the squat exercise, and increased the quality of repetitions performed.

Hoffman et al (2009). "Effect of Betaine Supplementation on Power Performance and Fatigue." Journal of the International Society of Sports Nutrition 6(1): 7.

Betaine and Muscle

This paper helps explain the positive effects seen on muscle by betaine.

Ortiz-Costa et al (2008) "Betaine protects urea-induced denaturation of myosin subfragment-1."

Myosin belongs to a family of motor proteins that transform the chemical energy of ATP into mechanical work.

Urea, a waste-product of protein metabolism, can increase due to exercise. Urea alters the structure of skeletal muscle myosin subfragment-1 (the catalytic domain), thereby inhibiting ATPase activity and reducing energy production.

Betaine, acting as a 'counteracting' osmolyte, was able to protect against these changes in structure, thereby preserving the enzyme’s catalytic activity.

Papers 2003-7:

Brigotti et al (2003). "Effects of osmolarity, ions and compatible osmolytes on cell-free protein synthesis." Biochem. J. 369(2): 369-374.