Insect Protein and Muscle Protein Signaling

Study Overview
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Brief Summary
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Randomized parallel study investing the effect of intake of different protein sources (whey, insect and pea) on the muscle protein synthesis. Activation of the signaling pathway leading to muscle protein synthesis is investigated by western blotting and Real time quantitative polymerase chain reaction (qPCR or PCR). Urine, blood and muscle is moreover investigated by metabolomics analysis.

Detailed Description
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Background: Sarcopenia is a syndrome highly prevalent in the older population, characterized by muscle loss and a decrease in muscle strength. This leads to loss of physical function, decreased life quality and well-being and an increased risk of early death. Research has shown that both ingestion of protein and physical activity is able to diminish the loss of muscle mass during aging and thereby reduce the prevalence of sarcopenia and diminish the consequence of the disease. This study will investigate if more sustainable protein sources of good quality can increase the muscle protein synthesis and thereby prevent sarcopenia. Aim: The aim of this project is to investigate the effects of insect protein on the stimulation of muscle protein synthesis. The effects of protein will both be studied alone and in combination with exercise in perspective of preventing loss of both muscle mass and muscle strength. The potential of insect protein will be elucidated by comparison with other alternative plant-based protein sources in this case pea protein. In addition, whey protein will be included as a positive control as it may be regarded as the most established dietary protein source combatting muscle loss. Method: Young(18-30 years old) healthy men (n=60) are randomized in 3 groups to ingest either insect protein, pea protein or whey protein. Urine are collected 24 hours prior to the experiment while a blood sample and a muscle biopsy is collected at the beginning of the study. The subjects are instructed to perform one-leg exercise (knee extension, 5 sets of 10 repetitions, 10 repetitions-maximum) after which the subjects ingest the assigned protein bolus. Urine and blood samples are collected in the following hours and one muscle biopsy is collected from both the exercised and the non-exercised leg at 3 hours after protein ingestion. The expression of messenger RNA (mRNA) and protein related to the mTORC pathway in the muscle is investigated by qPCR and western blotting. In addition, metabolites in urine, blood plasma and muscle tissue are investigated by metabolomics analysis.

Official Title
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Insect Protein - a New and Sustainable Protein Source to Maintain and Build Muscle

Conditions
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Sarcopenia

Intervention / Treatment
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* Dietary Supplement: insect, pea or whey protein


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: male 18-30 year able to read and understand danish Exclusion Criteria: smoking resistance training more than 1 time pr month endurance training more than 2 hours pr week Allergy toward shellfish, house dust mites or the protein sources. knee problems effecting the exercise weight change more than 5 kg the last half year medicine which can effect the protein turnover Disease in joint, or muscle as well as metabolic diseases.

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 30 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: Prevention
Allocation: Randomized
Intervention Model: Parallel Assignment
Interventional Model Description: A parallel 3 armed study, each arm with a different protein type
Masking: Double


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Insect protein<br>The participants are are given insect protein | Dietary Supplement: insect, pea or whey protein<br>* The protein powder is dissolved in 400 mL water and given relative to lean body mass, 0.25 g protein pr kg lean body mass.<br>|
| Experimental: Pea protein<br>The participants are are given pea protein | Dietary Supplement: insect, pea or whey protein<br>* The protein powder is dissolved in 400 mL water and given relative to lean body mass, 0.25 g protein pr kg lean body mass.<br>|
| Experimental: Whey protein<br>The participants are are given whey protein | Dietary Supplement: insect, pea or whey protein<br>* The protein powder is dissolved in 400 mL water and given relative to lean body mass, 0.25 g protein pr kg lean body mass.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Expression and phosphorylation of signaling proteins and their mRNA by Western Blotting and PCR | Change in expression and phosphorylation of signaling proteins and the mRNA related the muscle protein synthesis. | 4 hours | 
| Blod and urine metabolism by NMR | Both blod and urine are analysed by nuclear magnetic resonance (NMR) metabolomics to enlighten changes in the metabolites. | 4 hours | 
| Muscle metabolism by NMR | The muscle metabolites are quantified before intervention, after protein and after protein end exercise. | 3 hours | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| BMI | Control parameter between groups. Measured before the study day. | 1 hour | 
| General physical activity | in the last 3 month. Self-reported through a questionnaire. | 3 months | 
| Blood glucose | Blood glucose measured at 7 time points over the 4 hour period. | 4 hours | 
| Blood Insulin | Blood insulin is measured at 7 time points over the 4 hour period. | 4 hours | 
| Food intake, calories and distribution of macro nutrients. | Control parameter between groups. Self-reported diet report. | 1 day | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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mTORC metabolomics,, Western blotting, Real time quantitative polymerase chain reaction, Nuclear magnetic resonance, Liquid chromatography-mass spectrometry