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The Role of Muscle Protein Breakdown in the Regulation of Muscle Quality in Frail Elderly Individuals

Not Applicable
Completed
Conditions
Sarcopenia
Interventions
Other: Strength training
Dietary Supplement: Protein supplementation
Registration Number
NCT03326648
Lead Sponsor
Truls Raastad
Brief Summary

The purpose of this study is to investigate mechanisms underlying the reduction in muscle quality (the ratio between muscle strength and muscle size) with aging, and to investigate how these factors are affected by strength training and protein supplementation. It is already established that muscle quality defined as the ratio between the strength and the size of a muscle is improved with strength training, even in frail elderly individuals. However, the relative contribution of factors such as activation level, fat infiltration, muscle architecture and single fiber function is unknown. The main focus of this study is to investigate the relationship between muscle quality and muscle protein breakdown, as insufficient degradation of proteins is hypothesized to negatively affect muscle quality.

Detailed Description

Aging is associated with impaired skeletal muscle function. This is evident not only by a reduced capacity to generate force and power at the whole muscle level, but also by a decline in individual muscle fiber contraction velocity and force generation. Combined with muscle atrophy, these changes lead to reduced muscle strength and quality and loss off physical function with age. Clinically, muscle quality may be a better indicator of overall functional capacity than absolute muscle strength. Thus, identifying the mechanisms underlying the age-related loss of muscle quality is of high relevance for the prevention of functional impairment with aging. The explanation for the loss of muscle quality with aging seems to be multifactorial, with alterations in voluntary muscle activation, muscle architecture, fat infiltration and impaired contractile properties of single muscle fibers being likely contributors. Single fiber specific force seems to be related to myosin heavy chain (MHC) content, which is thought to reflect the number of available cross-bridges. The reduction of single fiber specific force with aging may thus be a consequence of reduced synthesis of MHC and/or increased concentration of non-contractile tissue (e.g. intramyocellular lipids).

Some studies in mice also indicate attenuated activity in some of the pathways responsible for degradation of muscle proteins with aging (especially autophagy). As a result, damaged proteins and organelles are not removed as effectively as they should, which could ultimately compromise the muscle's ability to produce force. In addition, reduced efficiency of mitophagy and lipophagy (two specific forms of autophagy), may indirectly affect single fiber specific force, through oxidative damage by reactive oxygen species (ROS) and increased levels of intramyocellular lipids, respectively. Although animal studies indicate attenuated autophagic function, exercise seems to restore the activity in this pathway. Whether this also is the case in humans is unknown. Thus, the purpose of this study is to investigate how the different factors contributing to reduced muscle quality in frail elderly individuals, with emphasis on the relationship between muscle quality and autophagy, may be counteracted by a specific strength training program targeting muscle quality and muscle mass.

In this randomized controlled trial the investigators will aim to recruit frail elderly individuals, as muscle quality is shown to be low in this population. As a consequence, the potential for improved muscle quality is expected to be large. Subjects will be randomized to two groups; one group performing strength training twice a week for 10 weeks in addition to receiving daily protein supplementation. The other group will only receive the protein supplement. Several tests will be performed before and after the intervention period, including a test day where a biopsy is obtained both at rest, and 2.5 hours following strength training + protein supplementation or protein supplementation only. This will provide information about the regulation of muscle protein breakdown in a resting state, following protein intake and following strength training in combination with protein intake. As this will be done both before and after the training period, it will also provide information on how long-term strength training affects the activity in these systems.

Recruitment & Eligibility

Status
COMPLETED
Sex
All
Target Recruitment
34
Inclusion Criteria
  • Age > 65
  • Frail or pre-frail according to the Fried Frailty Criteria or Short Physical Performance Battery (SPPB) score <6.
  • Mini Mental State Examination score > 18
Exclusion Criteria
  • Diseases or injuries contraindicating participation
  • Lactose intolerance
  • Allergy to milk
  • Allergy towards local anesthetics (xylocain)
  • Use of anticoagulants that cannot be discontinued prior to the muscle biopsy

Study & Design

Study Type
INTERVENTIONAL
Study Design
PARALLEL
Arm && Interventions
GroupInterventionDescription
Strength training + protein supplementProtein supplementationTwo sessions of strength training each week in addition to daily protein supplementation for 10 weeks.
Protein supplementProtein supplementationDaily protein supplementation for 10 weeks.
Strength training + protein supplementStrength trainingTwo sessions of strength training each week in addition to daily protein supplementation for 10 weeks.
Primary Outcome Measures
NameTimeMethod
Single fiber specific forceChange from baseline at 10 weeks

A measure of muscle quality at the single fiber level. Biopsies obtained from m. Vastus Lateralis

Secondary Outcome Measures
NameTimeMethod
Blood plasma Hemoglobin A1c (HbA1c)Change from baseline at 10 weeks

Fasted

Phosphorylation status and total level of ribosomal protein S6 kinase beta-1(P70S6K)Before and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

Level/cellular location of ubiquitin (Ub)Before and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

hepatocyte growth factor (HGF) mRNABefore and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

myostatin (MSTN) mRNABefore and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

p62/Sequestosome-1 mRNABefore and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

Microtubule-associated protein 1A/1B-light chain 3 (LC3) mRNABefore and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

PTEN-induced putative kinase 1 (PINK1) mRNABefore and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

TNF receptor associated factor 6 (TRAF6) mRNABefore and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

Intramyocellular lipidsChange from baseline at 10 weeks

Oil-Red-O staining of muscle sections. Biopsy from m. Vastus Lateralis analyzed by immunohistochemistry

Muscle fiber cross-sectional areaChange from baseline at 10 weeks

Biopsy from m. Vastus Lateralis analyzed by immunohistochemistry

E3 ubiquitin-protein ligase TRIM63 (TRIM63) mRNABefore and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

MyonucleiChange from baseline at 10 weeks

Biopsy from m. Vastus Lateralis analyzed by immunohistochemistry

Myonuclei locationChange from baseline at 10 weeks

Biopsy from m. Vastus Lateralis analyzed by confocal microscopy

Amount of mitochondriaChange from baseline at 10 weeks

Biopsy from m. Vastus Lateralis analyzed by confocal microscopy

Location of mitochondriaChange from baseline at 10 weeks

Biopsy from m. Vastus Lateralis analyzed by confocal microscopy

insulin-like growth factor I (IGF1) mRNABefore and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

Muscle activationChange from baseline at 10 weeks

Voluntary activation level during a maximal isometric knee extension using the interpolated twitch technique

Fat massChange from baseline at 10 weeks

Measured by a Dual-energy X-ray absorptiometry (DXA) scan

Bone mineral densityChange from baseline at 10 weeks

Measured by a Dual-energy X-ray absorptiometry (DXA) scan

Fractional Breakdown RateMeasured over the last 14 days of the intervention period

Measurement of fractional breakdown rate by the use of orally provided Deuterium Oxide, biopsies and blood samples

Phosphorylation status and total level of eukaryotic translation initiation factor 4E-binding protein 1 (4EBP-1)Before and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

Level/cellular location of muscle RING-finger protein-1 (Murf-1)Before and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

Blood serum glucoseChange from baseline at 10 weeks

Fasted

Lean massChange from baseline at 10 weeks

Measured by a Dual-energy X-ray absorptiometry (DXA) scan

Fat infiltration of m. quadricepsChange from baseline at 10 weeks

Fat infiltration of m. quadriceps measured by a Computed Tomography scan

m. Vastus Lateralis thicknessChange from baseline at 10 weeks

Measured by ultrasound

Habitual gait velocityChange from baseline at 10 weeks

Time (sec) to walk 6 meters at habitual gait velocity

Maximal gait velocityChange from baseline at 10 weeks

Time (sec) to walk 6 meters as fast as possible

Level/cellular location of Lysosome-associated membrane glycoprotein 2 (LAMP2)Before and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

Blood serum insulinChange from baseline at 10 weeks

Fasted

Blood serum High-density lipoproteins (HDL)Change from baseline at 10 weeks

Fasted

Blood serum C-reactive protein (CRP)Change from baseline at 10 weeks

Fasted

Muscle strength of m. quadricepsChange from baseline at 10 weeks

Maximal isometric and dynamic muscle strength of m. quadriceps

Blood serum Low-density lipoproteins (LDL)Change from baseline at 10 weeks

Fasted

Myonuclei numberChange from baseline at 10 weeks

Biopsy from m. Vastus Lateralis analyzed by confocal microscopy

Muscle satellite cellsChange from baseline at 10 weeks

Biopsy from m. Vastus Lateralis analyzed by immunohistochemistry

Chair stand performanceChange from baseline at 10 weeks

Time (sec) to stand up from a chair five times

Level/cellular location of p62/Sequestosome-1Before and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

Phosphorylation status and total level of eukaryotic elongation factor 2 (eEF-2)Before and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

forkhead box protein O1 (FOXO1) mRNA mRNABefore and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

Muscle size of m. quadricepsChange from baseline at 10 weeks

Cross-sectional area of m. quadriceps measured by a Computed Tomography scan

Level/cellular location of Microtubule-associated protein 1A/1B-light chain 3 (LC3)Before and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

Level/cellular location of forkhead box O3 (FOXO3a)Before and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

Blood serum TriglyceridesChange from baseline at 10 weeks

Fasted

forkhead box protein O3 (FOXO3A) mRNABefore and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

muscle RING-finger protein-1 (Murf-1) protein 1 (4EBP-1) mRNABefore and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

Atrogin1 mRNABefore and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

BCL2/adenovirus E1B interacting protein 3 (BNIP3) mRNABefore and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

transcription factor EB (Tfeb) mRNABefore and 2.5 hours after acute training session both at baseline and after 10 weeks

Biopsies from m. Vastus Lateralis analyzed by western blot

Muscle fiber type distributionChange from baseline at 10 weeks

Biopsy from m. Vastus Lateralis analyzed by immunohistochemistry

Trial Locations

Locations (1)

Norwegian School of Sport Sciences

🇳🇴

Oslo, Norway

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