Heart and Vessels

, Volume 31, Issue 10, pp 1685–1695 | Cite as

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

  • Toshiaki NakajimaEmail author
  • Tomohiro Yasuda
  • Seiichiro Koide
  • Tatsuya Yamasoba
  • Syotaro Obi
  • Shigeru Toyoda
  • Yoshiaki Sato
  • Teruo Inoue
  • Yutaka Kano
Original Article


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.


Blood flow restriction Mammalian target of rapamycin (mTOR) Microvascular pO2 Muscle atrophy Myostatin p70 S6-kinase Rat skeletal muscle Hypoxia 



Restriction of muscle blood flow


Repetitive restriction of muscle blood flow


Microvascular oxygen pressure


Hypoxia-inducible factor-1α


Mammalian target of rapamycin


p70 S6-kinase


Extracellular signal-regulated kinase 1/2


Forkhead box O3A


Muscle ring finger-1


Vascular endothelial growth factor



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).

Compliance with ethical standards

Conflict of interest

There is no conflict of interests to disclose.


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Copyright information

© Springer Japan 2016

Authors and Affiliations

  • Toshiaki Nakajima
    • 1
    Email author return OK on get
  • Tomohiro Yasuda
    • 3
  • Seiichiro Koide
    • 4
  • Tatsuya Yamasoba
    • 5
  • Syotaro Obi
    • 2
  • Shigeru Toyoda
    • 1
  • Yoshiaki Sato
    • 6
  • Teruo Inoue
    • 1
  • Yutaka Kano
    • 4
  1. 1.Department of Cardiovascular MedicineDokkyo Medical University and Heart Center, Dokkyo Medical University HospitalTochigiJapan
  2. 2.Department of Cardiovascular Medicine and Research Support CenterDokkyo Medical UniverasityTochigiJapan
  3. 3.Seirei Christopher UniversityShizuokaJapan
  4. 4.Department of Engineering Science, Bioscience and Technology ProgramUniversity of Electro-CommunicationsTokyoJapan
  5. 5.Department of OtolaryngologyUniversity of TokyoTokyoJapan
  6. 6.Department of Basic Sciences in MedicineKaatsu International UniversityBattaramullaSri Lanka

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