Anabolic Effects of Whey and Casein After Strength Training in Young and Elderly

Study Overview
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Brief Summary
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The aim of this study is to investigate the acute anabolic effects of native whey, whey protein concentrate 80 (WPC-80) and milk after a bout of strength training in young and elderly. The investigators hypothesize that native whey will give a greater stimulation of muscle protein synthesis and intracellular anabolic signaling than WPC-80, and that WPC-80 will give a stronger stimulus than milk.

Detailed Description
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Increasing or maintaining muscle mass is of great importance for populations ranging from athletes to patients and elderly. Resistance exercise and protein ingestion are two of the most potent stimulators of muscle protein synthesis. Both the physical characteristic of proteins (e.g. different digestion rates of whey and casein) and the amino acid composition, affects the potential of a certain protein to stimulate muscle protein synthesis. Given its superior ability to rapidly increase blood leucine concentrations to high levels, whey is often considered the most potent protein source to stimulate muscle protein synthesis. Native whey protein is produced by filtration of unprocessed milk. Consequently, native whey has different characteristics than WPC-80, which is exposed to heating and acidification. Because of the direct filtration of unprocessed milk, native whey is a more intact protein compared with WPC-80. Of special interest is the higher amounts of the highly anabolic amino acid leucine in native whey. The higher levels of leucine can be of great interest for elderly individuals as some studies in elderly has shown an anabolic resistance to the effects of protein feeding and strength training. By increasing levels of leucine one might overcome this anabolic resistance in the elderly. The aim of this double-blinded, randomized, partial cross-over study is to compare the acute fractional protein synthesis and intracellular signaling response to a bout of strength training and intake of 20 grams of protein from either native whey, whey protein concentrate 80 or milk, in young and old individuals. Furthermore, the investigators wil investigate fractional protein breakdown, markers of protein breakdown, amino acid concentrations in blood. The investigators hypothesize that native whey will induce a greater anabolic response than whey protein concentrate 80, and that whey protein concentrate 80 will give a stronger anabolic response than milk.

Official Title
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Effects of Whey and Casein Supplementation on Acute Anabolic Responses in Muscle After Strength Training in Young and Elderly

Conditions
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Healthy, Young, Elderly

Intervention / Treatment
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* Other: Strength training
* Dietary Supplement: Milk 1%
* Dietary Supplement: Whey protein concentrate 80
* Dietary Supplement: Native whey


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Healthy in the sense that they can conduct training and testing Able to understand Norwegian language written and oral Between 18 and 45, or above 70 years of age Exclusion Criteria: Diseases or injuries contraindicating participation Use of dietary supplements (e.g. proteins, vitamins and creatine) Lactose intolerance Allergy to milk Allergy towards local anesthetics (xylocain)

Ages Eligible for Study
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Minimum Age: 18 Years

Sexes Eligible for Study
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All

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: Basic Science
Allocation: Randomized
Intervention Model: Crossover Assignment
Masking: Triple


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Placebo Comparator: Milk<br>Participants performed a bout of strength training and consumed 20g of milk protein | Other: Strength training<br> <br> Dietary Supplement: Milk 1%<br> <br> |
| Experimental: Whey protein concentrate 80<br>Participants performed a bout of strength training and consumed 20g of whey protein concentrate 80 | Other: Strength training<br> <br> Dietary Supplement: Whey protein concentrate 80<br> <br> |
| Experimental: Native whey<br>Participants performed a bout of strength training and consumed 20g of native whey | Other: Strength training<br> <br> Dietary Supplement: Native whey<br> <br> |

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Mixed muscle fractional synthetic rate | A continous infusion of a stable isotope (phe D5) is used to measure incorporation of tracer into muscle (biopsies from m. vastus lateralis) | Three to one hours prior to a bout of strength training and protein consumption | 
| Mixed muscle fractional synthetic rate | A continous infusion of a stable isotope (phe D5) is used to measure incorporation of tracer into muscle (biopsies from m. vastus lateralis) | One to five hours after a bout of strength training and protein consumption | 
| Mixed muscle fractional synthetic rate | Two boluses of tracer (phe13C6 and phe15N) was used to measure incorporation of tracer into muscle (biopsies from m. vastus lateralis) | From three to five hours after a bout of strength training and protein consumption | 
| Mixed muscle fractional breakdown rate | Two boluses of tracer (phe13C6 and phe15N) was used to measure the dilution of tracer in muscle (biopsies from m. vastus lateralis) | From three to five hours after a bout of strength training and protein consumption | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Ratio of phosphorylated to total ribosomal protein S6 kinase beta-1(P70S6K) change from baseline | Biopsies from m. Vastus Lateralis was analyzed by western blot | 30 min before, 1, 2.5 and 5 hours after training and protein intake | 
| Phosphorylation of phosphorylated to total eukaryotic elongation factor 2 (eEF-2) change from baseline | Biopsies from m. Vastus Lateralis was analyzed by western blot | 30 min before, 1, 2.5 and 5 hours after training and protein intake | 
| Phosphorylation of phosphorylated to total eukaryotic translation initiation factor 4E-binding protein 1 (4EBP-1) change from baseline | Biopsies from m. Vastus Lateralis was analyzed by western blot | 30 min before, 1, 2.5 and 5 hours after training and protein intake | 
| Intracellular translocation of forkhead box O3 (FOXO3a) change from baseline | Biopsies from m. Vastus Lateralis was analyzed by western blot | 30 min before, 1, 2.5 and 5 hours after training and protein intake | 
| Intracellular translocation of muscle RING-finger protein-1 (Murf-1) change from baseline | Biopsies from m. Vastus Lateralis was analyzed by western blot | 30 min before, 1, 2.5 and 5 hours after training and protein intake | 
| Intracellular translocation of Atrogin1 change from baseline | Biopsies from m. Vastus Lateralis was analyzed by western blot | 30 min before, 1, 2.5 and 5 hours after training and protein intake | 
| Ubiquitin | Biopsies from m. Vastus Lateralis was analyzed by western blot | 30 min before, 1, 2.5 and 5 hours after training and protein intake | 
| Plasma amino acid concentration |  | 180 and 60 min before, and 45, 60, 75, 120, 160, 180, 200, 220 and 300 min after training and protein intake | 
| Muscle force generating capacity change from baseline | Measured as unilateral isometric knee extension force (Nm) with 90° in the hip and knee joints. | 15 min before, 15 and 300 min after, and 24 hours after training and protein intake | 
| Plasma glucose |  | 180 and 60 min before, and 45, 60, 75, 120, 160, 180, 200, 220 and 300 min after training and protein intake | 
| Plasma insulin |  | 180 and 60 min before, and 45, 60, 75, 120, 160, 180, 200, 220 and 300 min after training and protein intake | 
| Serum urea |  | 180 and 60 min before, and 60, 10, 180 and 300 min after training and protein intake | 
| Serum ureic acid |  | 180 and 60 min before, and 60, 10, 180 and 300 min after training and protein intake | 
| Serum creatine kinase |  | 180 and 60 min before, and 60, 10, 180 and 300 min after training and protein intake | 
| Change in ATP-binding cassette transporter (ABCA1) messenger ribonucleic acid (mRNA) |  | 1 hour after training and protein intake | 
| Change in ABCA1 mRNA |  | 5 hous after training and protein intake | 
| Change in BRCA1-A complex subunit Abraxas (ABRA1) mRNA |  | 1 hour after training and protein intake | 
| Change in ABRA1 mRNA |  | 5 hours after training and protein intake | 
| Change in alfa-actin (ACTA1) mRNA |  | 1 hour after training and protein intake | 
| Change in ACTA1 mRNA |  | 5 hours after training and protein intake | 
| Change in C-C motif chemokine 2 (CCL2) mRNA |  | 1 hour after training and protein intake | 
| Change in CCL2 mRNA |  | 5 hours after training and protein intake | 
| Change in C-C motif chemokine 3 (CCL3) mRNA |  | 1 hour after training and protein intake | 
| Change in CCL3 mRNA |  | 5 hours after training and protein intake | 
| Change in C-C motif chemokine 5 (CCL5) mRNA |  | 1 hour after training and protein intake | 
| Change in CCL5 mRNA |  | 5 hours after training and protein intake | 
| Change in C-C motif chemokine 8 (CCL8) mRNA |  | 1 hour after training and protein intake | 
| Change in CCL8 mRNA |  | 5 hours after training and protein intake | 
| Change in platelet glycoprotein 4 (CD36) mRNA |  | 1 hour after training and protein intake | 
| Change in CD36 mRNA |  | 5 hours after training and protein intake | 
| Change in cholesterol 25-hydroxylase (CH25H) mRNA |  | 1 hour after training and protein intake | 
| Change in CH25H mRNA |  | 5 hours after training and protein intake | 
| Change in granulocyte colony-stimulating factor (CSF3) mRNA |  | 1 hour after training and protein intake | 
| Change in CSF3 mRNA |  | 5 hours after training and protein intake | 
| Change in C-X-C motif chemokine 16 (CXCL16) mRNA |  | 1 hour after training and protein intake | 
| Change in CXCL16 mRNA |  | 5 hours after training and protein intake | 
| Change in F-box only protein 32 (FBXO32) mRNA |  | 1 hour after training and protein intake | 
| Change in FBXO32 mRNA |  | 5 hours after training and protein intake | 
| Change in growth-regulated alpha protein (CXCL1) mRNA |  | 1 hour after training and protein intake | 
| Change in CXCL1 mRNA |  | 5 hours after training and protein intake | 
| Change in matrix metalloproteinase-9 (MMP9) mRNA |  | 1 hour after training and protein intake | 
| Change in MMP9 mRNA |  | 5 hours after training and protein intake | 
| Change in forkhead box protein O1 (FOXO1) mRNA |  | 1 hour after training and protein intake | 
| Change in FOXO1 mRNA |  | 5 hours after training and protein intake | 
| Change in forkhead box protein O3 (FOXO3A) mRNA |  | 1 hour after training and protein intake | 
| Change in FOXO3A mRNA |  | 5 hours after training and protein intake | 
| Change in hepatocyte growth factor (HGF) mRNA |  | 1 hour after training and protein intake | 
| Change in HGF mRNA |  | 5 hours after training and protein intake | 
| Change in insulin-like growth factor I (IGF1) mRNA |  | 1 hour after training and protein intake | 
| Change in IGF1 mRNA |  | 5 hours after training and protein intake | 
| Change in interleukin-10 (IL10) mRNA |  | 1 hour after training and protein intake | 
| Change in IL10 mRNA |  | 5 hours after training and protein intake | 
| Change in interleukin-17D (IL17D) mRNA |  | 1 hour after training and protein intake | 
| Change in IL17D mRNA |  | 5 hours after training and protein intake | 
| Change in interleukin-1B (IL1B) mRNA |  | 1 hour after training and protein intake | 
| Change in IL1B mRNA |  | 5 hours after training and protein intake | 
| Change in interleukin-1 receptor antagonist protein (IL1RN) mRNA |  | 1 hour after training and protein intake | 
| Change in IL1RN mRNA |  | 5 hours after training and protein intake | 
| Change in interleukin-6 (IL6) mRNA |  | 1 hour after training and protein intake | 
| Change in IL6 mRNA |  | 5 hours after training and protein intake | 
| Change in interleukin-8 (IL8) mRNA |  | 1 hour after training and protein intake | 
| Change in IL8 mRNA |  | 5 hours after training and protein intake | 
| Change in transcription factor jun-B (JUNB) mRNA |  | 1 hour after training and protein intake | 
| Change in JUNB mRNA |  | 5 hours after training and protein intake | 
| Change in kit ligand (KITLG) mRNA |  | 1 hour after training and protein intake | 
| Change in KITLG mRNA |  | 5 hours after training and protein intake | 
| Change in myostatin (MSTN) mRNA |  | 1 hour after training and protein intake | 
| Change in MSTN mRNA |  | 5 hours after training and protein intake | 
| Change in myosin-1 (MYH1) mRNA |  | 1 hour after training and protein intake | 
| Change in MYH1 mRNA |  | 5 hours after training and protein intake | 
| Change in myosin-2 (MYH2) mRNA |  | 1 hour after training and protein intake | 
| Change in MYH2 mRNA |  | 5 hours after training and protein intake | 
| Change in myosin-7 (MYH7) mRNA |  | 1 hour after training and protein intake | 
| Change in MYH7 mRNA |  | 5 hours after training and protein intake | 
| Change in oxysterols receptor LXR-alpha (NR1H3) mRNA |  | 1 hour after training and protein intake | 
| Change in NR1H3 mRNA |  | 5 hours after training and protein intake | 
| Change in nuclear receptor subfamily 4 group A member 3 (NR4A3) mRNA |  | 1 hour after training and protein intake | 
| Change in NR4A3 mRNA |  | 5 hours after training and protein intake | 
| Change in peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PPARGC1A) mRNA |  | 1 hour after training and protein intake | 
| Change in PPARGC1A mRNA |  | 5 hours after training and protein intake | 
| Change in prostaglandin G/H synthase 2 (PTGS2) mRNA |  | 1 hour after training and protein intake | 
| Change in PTGS2 mRNA |  | 5 hours after training and protein intake | 
| Change in proton-coupled amino acid transporter 1 (SLC36A1) mRNA |  | 1 hour after training and protein intake | 
| Change in SLC36A1 mRNA |  | 5 hours after training and protein intake | 
| Change in sodium-coupled neutral amino acid transporter 2 (SLC38A2) mRNA |  | 1 hour after training and protein intake | 
| Change in SLC38A2 mRNA |  | 5 hours after training and protein intake | 
| Change in 4F2 cell-surface antigen heavy chain (SLC3A2) mRNA |  | 1 hour after training and protein intake | 
| Change in SLC3A2 mRNA |  | 5 hours after training and protein intake | 
| Change in large neutral amino acids transporter small subunit 1 (SLC7A5) mRNA |  | 1 hour after training and protein intake | 
| Change in SLC7A5 mRNA |  | 5 hours after training and protein intake | 
| Change in toll-like receptor 2 (TLR2) mRNA |  | 1 hour after training and protein intake | 
| Change in TLR2 mRNA |  | 5 hours after training and protein intake | 
| Change in tumor necrosis factor (TNF) mRNA |  | 1 hour after training and protein intake | 
| Change in TNF mRNA |  | 5 hours after training and protein intake | 
| Change in E3 ubiquitin-protein ligase TRIM63 (TRIM63) mRNA |  | 1 hour after training and protein intake | 
| Change in E3 ubiquitin-protein ligase TRIM63 (TRIM63) mRNA |  | 5 hours after training and protein intake |