Potential Synergistic Effect of Combined Blood Flow Restriction Training and Betaine Supplementation on Skeletal Muscle

Study Overview
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Brief Summary
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The purpose of the study is to determine whether there is a synergistic effect via combining both low-load blood flow restriction (BFR) training and betaine supplementation loading (6g/day for 14 days) on skeletal muscle anabolic signaling pathways that is mediated by enhancements in intracellular water. These effects are proposed to be greater than either BFR training or betaine supplementation alone or compared to control conditions (high-load non-occluded and/or placebo supplementation).

Detailed Description
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The purpose of the investigation is to determine whether the combination of blood flow restriction (BFR) training and betaine supplementation can synergistically augment phosphorylated targets associated with mechanotransduction and/or muscle protein synthesis relative to either modality alone and compared against control conditions (standard high-intensity resistance training and placebo supplementation) in healthy young males. Secondly, the investigators aim to determine if any potential synergistic effects are mediated by enhanced intracellular fluid volumes, as determined by the changes in water content between hydrated and dehydrated muscle samples, as well as through changes in both muscle and serum betaine concentrations. Finally, the investigators aim to assess differences in the aforementioned interventions on specific gene targets, the betaine/γ-aminobutyric acid transporter, myosin heavy chain I, IIa, and IIx lactate dehydrogenase A. Therefore, the specific aims of this study are to determine in healthy, young males: 1) whether combined BFR training and betaine supplementation significantly augment mechanotransductive growth-associated post-translational protein modifications via extra-to-intracellular fluid flux, alongside 2) potentially altered gene expression that otherwise characterizes phenotypical/biochemical changes in skeletal muscle. The specific aims of the study are to determine whether: The combination of BFR training and betaine supplementation demonstrates significantly greater phosphorylated FAK, ERK1/2, IRS1, and p70S6K, commensurate with greater wet-to-dry hydration changes, relative to any other combinations between BFR training, standard high-load training, betaine supplementation, and/or placebo ingestion. The combination of BFR training and betaine supplementation will result in increased MYH2 gene expression, alongside decreases in MYH7 and MYH1 expression. Furthermore, this combination will also result in the highest degree of HIF-1 and Ldha, as well as the lowest BGT-1 gene expression relative to baseline levels. The combination of BFR training and betaine supplementation will result in a higher load-volume accumulated relative to BFR-alone, and will not be statistically different than high-load-placebo training. Therein, the high-load-betaine group will have the greatest load-volume amidst any other combination of conditions.

Official Title
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The Potential Synergistic Effect of Combined Blood Flow Restriction Training and Betaine Supplementation on Skeletal Muscle Mechanotransduction-Associated Cell Signaling

Conditions
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Blood Flow Restriction Training, Betaine Supplementation

Intervention / Treatment
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* Dietary Supplement: Betaine
* Other: Placebo


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Only participants considered low risk for cardiovascular disease with no contraindications to exercise as outlined by the ACSM Have not consumed any nutritional supplements (aside from a multi-vitamin) one month prior to investigation Blood pressure <140/90mmHg Resting heart rate <90bpm- Exclusion Criteria: sedentary individual as defined by the ACSM guidelines. inadequate resistance training experience (<12 months, <3x/week) vegetarian, vegan, or have dietary restrictions or supplements that potentially affect betaine metabolism. allergy to topical anesthetics. known metabolic or cardiovascular disorder including heart disease, arrhythmias, diabetes, thyroid disease, or hypogonadism. genetic disorders/polymorphisms that would act as direct contraindications to betaine supplementation (i.e. methyltetrahydrofolate reductase, hyperhomocysteinemia, etc.) bleeding disorder, history of pulmonary disease, hypertension, hepatorenal disease, musculoskeletal disorders, neuromuscular/ neurological diseases, autoimmune disease, cancer, peptic ulcers, anemia, or chronic infection (e.g., HIV). resting systolic/diastolic blood pressure and heart rate of more than 140/90 mmHg and 90, respectively. taking any blood thinning (e.g., warfarin, Jantoven, etc.), heart, pulmonary, thyroid, anti-hyperlipidemic, hypoglycemic, anti-hypertensive, endocrinologic (e.g, thyroid, insulin, etc), emotional/psychotropic (e.g., Prednisone, Ritalin, Adderall), or neuromuscular/neurological medications. taking anabolic androgenic steroids within the past year.

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 35 Years

Sexes Eligible for Study
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Male

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Other
Allocation: Randomized
Intervention Model: Crossover Assignment
Masking: Single


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Placebo Comparator: Placebo<br>3g/ twice daily (separated by  12 hours) cellulose placebo for 14 days | Other: Placebo<br>* 3g/ twice daily (separated by  12 hours) cellulose placebo<br>|
| Experimental: Betaine Supplementation<br>3g/twice daily (separated by  12 hours) betaine anhydrous for 14 days | Dietary Supplement: Betaine<br>* 3g/ twice daily (separated by  12 hours) betaine anhydrous<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| serum/muscle phosphorylated FAK | Focal adhesion kinase | 3 hours following exercise cessation | 
| insulin receptor substrate 1 | signalling adapter protein | 3 hours following exercise cessation | 
| gene expression of HIF-1 | genes related to skeletal muscle adaptation | 3 hours following exercise cessation | 
| serum and muscle betaine concentrations | measure related to the supplementation protocol | 3 hours following exercise cessation | 
| gene expression BGT-1 | genes related to skeletal muscle adaptation | 3 hours following exercise cessation | 
| gene expression MHC | genes related to skeletal muscle adaptation | 3 hours following exercise cessation | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| pre-to-post exercise set tissue hydration | hydration status of the participant via multi-frequency bioelectrical impedance | both pre and immediately post exercise session | 
| capillary blood lactate concentrations | blood lactate levels of the participant | both pre and immediately post exercise session | 
| set-to-failure repetition number | as related to the exercise protocol | immediately post exercise session | 
| exercise condition total load-volume. | as related to the exercise protocol and calculated at termination of the exercise session | immediately post exercise session |