Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Effect of Heat-Killed Enterococcus faecalis, EF-2001 on C2C12 Myoblast Damage Induced by Oxidative Stress and Muscle Volume Decreased by Sciatic Denervation in C57BL/6 Mice

산화스트레스에 의해 유도된 C2C12 근세포 손상과, 신경절제에 의해 근감소가 유도된 C57BL/6 마우스에서 열처리 사균체 엔테로코커스 패칼리스 EF-2001의 효과

  • Chang, Sang-Jin (Division of Biological Science and Technology, Yonsei University) ;
  • Lee, Myung-Hun (Division of Biological Science and Technology, Yonsei University) ;
  • Kim, Wan-Joong (Division of Biological Science and Technology, Yonsei University) ;
  • Chae, Yuri (Division of Biological Science and Technology, Yonsei University) ;
  • Iwasa, Masahiro (Division of Biological Science and Technology, Yonsei University) ;
  • Han, Kwon-Il (Division of Biological Science and Technology, Yonsei University) ;
  • Kim, Wan-Jae (Research & Development Center, Korea BeRM Co. Ltd.) ;
  • Kim, Tack-Joong (Division of Biological Science and Technology, Yonsei University)
  • 장상진 (연세대학교 생명과학기술학부) ;
  • 이명헌 (연세대학교 생명과학기술학부) ;
  • 김완중 (연세대학교 생명과학기술학부) ;
  • 채유리 (연세대학교 생명과학기술학부) ;
  • 이와사 마사히로 (연세대학교 생명과학기술학부) ;
  • 한권일 (연세대학교 생명과학기술학부) ;
  • 김완재 (주식회사 한국베름 연구개발센터) ;
  • 김택중 (연세대학교 생명과학기술학부)
  • Received : 2018.12.06
  • Accepted : 2018.12.28
  • Published : 2019.02.28
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Abstract

Muscle dysfunction may arise from skeletal muscle atrophy caused by aging, injury, oxidative stress, and hereditary disease. Powdered heat-killed Enterococcus faecalis (EF-2001) has anti-allergy, anti-inflammatory, and anti-tumor effects. However, its antioxidant and anti-atrophy effects are poorly characterized. In this study, we examined the effects of EF-2001 on muscle atrophy. To determine the protective effect of EF-2001 on oxidative stress, C2C12 myoblasts were treated with $H_2O_2$ to induce oxidative stress. This induced cell damage, which was reduced by treatment with EF-2001. The mechanism of EF-2001's effect was examined in response to oxidative stress. Treatment with EF-2001 reversed the expression of HSP70 and SOD1 proteins. Also, mRNA levels of Atrogin-1/MAFbx and MuRF1 increased under oxidative stress conditions but decreased following EF-2001 treatment. To evaluate muscle volume, two and three dimensional models of the muscles were analyzed using micro-CT. As expected, muscle volume decreased after sciatic denervation and recovered after oral administration of EF-2001. Therefore, EF-2001 is a candidate for the treatment of muscular atrophy, and future discovery of the additional effects of EF-2001 may yield further applications as a functional food with useful activities in various fields.

노화, 상해, 유전병 및 산화 스트레스와 같은 다양한 원인으로 인해 근육 위축을 유발한다. 그 동안의 연구에 의하면 열처리 사균체 엔테로코커스 패칼리스(EF-2001)는 항알레르기, 항염증 및 항종양 효과를 보였다. 그러나 항산화 및 항근위축에 대한 효과는 잘 알려져 있지 않고 있다. 본 연구에서는 EF-2001이 근육 위축에 미치는 영향을 연구 하였다. 산화 스트레스에 의한 EF-2001의 세포손상 보호 효과를 확인하기 위해 C2C12 근섬유 아세포는 H2O2로 처리되어, 산화 스트레스를 유도하여 세포 손상을 유발하였다. 그러나 EF-2001 처리로 인해 근세포 손상이 감소됨을 확인 하였다. 우리는 산화스트레스에 의한 EF-2001의 근세포손상의 감소 효과에 대한 메커니즘을 확인하였다. EF-2001는 산화스트레스로 유도된 근세포내의 HSP70 및 SOD1 단백질의 발현을 감소시켰다. 또한, 근세포내에 Atrogin-1/MAFbx 및 MuRF1의 mRNA 수준은 산화 스트레스 조건 하에서 증가하였으나, EF-2001에 의해 감소하였다. 더나아가 근감소를 유도한 좌골신경 절제 모델동물을 통한 근육량을 확인하기 위해 마이크로 CT를 활용해 2차원과 3차원으로 분석하였다. 근육량은 좌골 신경 절제 후에 감소하였고, EF-2001의 경구 투여에 의해 근육량이 회복되었다. 본 결과는 열처리 사균체 엔테로코커스 패칼리스인 EF-2001이 노인들에게 자주 발생하는 근감소증의 예방 및 개선할 수 있는 기능성 식품으로 다양한 분야에서 유용하게 활용 될 수 있음을 시사한다.

Keywords

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Fig. 2 Effect of EF-2001 on apoptosis in C2C12 myoblasts induced by oxidative stress.

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Fig. 3. Effect of EF-2001 on HSP70 and SOD1 protein expression in C2C12 myoblasts induced by oxidative stress.

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Fig. 4. Effect of EF-2001 on Atrogin-1/MAFbx and MuRF1 mRNA expression in C2C12 myoblasts induced by oxidative stress.

SMGHBM_2019_v29n2_215_f0005.png 이미지

Fig. 5. Effect of EF-2001 on muscle atrophy induced by sciatic denervation in C57BL/6 mice.

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Fig. 6. EF-2001 can restore muscle atrophy caused by oxidative stress in vitro and in vivo.

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Fig. 1. Effects of Effect of EF-2001 on the viability of C2C12 myoblasts induced by oxidative stress.

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