Holwerda · International journal of sport nutrition and exercise metabolism 2021 · randomized controlled trial · n=36

Exercise Plus Presleep Protein Ingestion Increases Overnight Muscle Connective Tissue Protein Synthesis Rates in Healthy Older Men.

Cited 20 times in the scientific literature.

Level 2 - randomized trial

Individual randomized controlled trial

PubMed 33588378 · doi:10.1123/ijsnem.2020-0222 · record verified 2026-08-29

What was done

Thirty-six healthy older men were randomly assigned to one of three groups (n = 12 each): ingestion of a nonprotein placebo before sleep (PLA), ingestion of 40 g of intrinsically labeled casein protein before sleep (PRO), or a single evening bout of resistance exercise followed by 40 g of presleep intrinsically labeled casein protein (EX+PRO). Stable isotope tracer infusions (L-[ring-2H5]-phenylalanine and L-[1-13C]-leucine) alongside blood and muscle tissue sampling were performed across overnight sleep to assess muscle connective tissue protein synthesis rates and dietary amino acid incorporation.

What was found

Presleep protein alone did not significantly increase overnight muscle connective tissue protein synthesis rates compared to placebo (0.060 ± 0.024%/hr in PRO vs. 0.049 ± 0.013%/hr in PLA; p = .73). In contrast, EX+PRO resulted in higher synthesis rates (0.095 ± 0.022%/hr) compared with both PLA and PRO (p < .01). Furthermore, prior exercise increased the incorporation of dietary protein-derived amino acids into muscle connective tissue protein (0.054 ± 0.009 mole percent excess in EX+PRO vs. 0.036 ± 0.013 in PRO; p < .01).

Why it matters

These findings demonstrate that presleep protein ingestion alone is insufficient to stimulate overnight muscle connective tissue protein synthesis in older adults, but combining prior resistance exercise with presleep protein effectively promotes connective tissue remodeling during sleep.

Limits

The study was restricted to a small cohort of healthy older men, limiting generalizability to women, younger individuals, or clinical populations. It measured only acute, single-night synthetic responses rather than long-term structural adaptations, and it lacked an exercise-only comparator group without protein.

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