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Repetitive restriction of muscle blood flow enhances mTOR signaling pathways in a rat model

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Abstract

Skeletal muscle is a plastic organ that adapts its mass to various stresses by affecting pathways that regulate protein synthesis and degradation. This study investigated the effects of repetitive restriction of muscle blood flow (RRMBF) on microvascular oxygen pressure (PmvO2), mammalian target of rapamycin (mTOR) signaling pathways, and transcripts associated with proteolysis in rat skeletal muscle. Eleven-week-old male Wistar rats under anesthesia underwent six RRMBF consisting of an external compressive force of 100 mmHg for 5 min applied to the proximal portion of the right thigh, each followed by 3 min rest. During RRMBF, PmvO2 was measured by phosphorescence quenching techniques. The total RNA and protein of the tibialis anterior muscle were obtained from control rats, and rats treated with RRMBF 0–6 h after the stimuli. The protein expression and phosphorylation of various signaling proteins were determined by western blotting. The mRNA expression level was measured by real-time RT-PCR analysis. The total muscle weight increased in rats 0 h after RRMBF, but not in rats 1–6 h. During RRMBF, PmvO2 significantly decreased (36.1 ± 5.7 to 5.9 ± 1.7 torr), and recovered at rest period. RRMBF significantly increased phosphorylation of p70 S6-kinase (p70S6k), a downstream target of mTOR, and ribosomal protein S6 1 h after the stimuli. The protein level of REDD1 and phosphorylation of AMPK and MAPKs did not change. The mRNA expression levels of FOXO3a, MuRF-1, and myostatin were not significantly altered. These results suggested that RRMBF significantly decreased PmvO2, and enhanced mTOR signaling pathways in skeletal muscle using a rat model, which may play a role in diminishing muscle atrophy under various conditions in human studies.

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Abbreviations

RMBF:

Restriction of muscle blood flow

RRMBF:

Repetitive restriction of muscle blood flow

PmvO2 :

Microvascular oxygen pressure

HIF-1α:

Hypoxia-inducible factor-1α

mTOR:

Mammalian target of rapamycin

p70S6k:

p70 S6-kinase

ERK1/2:

Extracellular signal-regulated kinase 1/2

FOXO3a:

Forkhead box O3A

MuRF-1:

Muscle ring finger-1

VEGF:

Vascular endothelial growth factor

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Acknowledgments

Dr. Yoshiaki Sato is an inventor of low-load resistance training with blood flow restriction, so-called Kaatsu training, and the owner of KAATSU Japan Co. Ltd. This work was supported by JSPS KAKENHI Grant Number 24300189 (to T.N.) and 25750288 (to T.Y.). This study was previously presented, in part, in abstract form at the Scientific Sessions, American Heart Association 2013 (Dallas, Texas, USA, 2013/11/16-11/20).

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Authors and Affiliations

  1. Department of Cardiovascular Medicine, Dokkyo Medical University and Heart Center, Dokkyo Medical University Hospital, 880 Kitakobayashi, Mibu, Tochigi, 321-0293, Japan

    Toshiaki Nakajima, Shigeru Toyoda & Teruo Inoue

  2. Department of Cardiovascular Medicine and Research Support Center, Dokkyo Medical Univerasity, Tochigi, Japan

    Syotaro Obi

  3. Seirei Christopher University, Shizuoka, Japan

    Tomohiro Yasuda

  4. Department of Engineering Science, Bioscience and Technology Program, University of Electro-Communications, Tokyo, Japan

    Seiichiro Koide & Yutaka Kano

  5. Department of Otolaryngology, University of Tokyo, Tokyo, Japan

    Tatsuya Yamasoba

  6. Department of Basic Sciences in Medicine, Kaatsu International University, Battaramulla, Sri Lanka

    Yoshiaki Sato

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  1. Toshiaki Nakajima
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  2. Tomohiro Yasuda
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Correspondence to Toshiaki Nakajima.

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Nakajima, T., Yasuda, T., Koide, S. et al. Repetitive restriction of muscle blood flow enhances mTOR signaling pathways in a rat model. Heart Vessels 31, 1685–1695 (2016). https://doi.org/10.1007/s00380-016-0801-6

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