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생쥐 근육세포에서 코티졸이 세포질세망 스트레스, 자연 세포사멸과 자가포식에 미치는 영향

Effects of Cortisol on Endoplasmic Reticulum-stress, Apoptosis, and Autophagy in Mouse Muscle C2C12 Cells

  • 신동현 (전북대학교 농업생명과학대학 동물생명공학과) ;
  • 김경환 (부산대학교 생명자원과학대학 동물생명자원과학과) ;
  • 이지현 (부산대학교 생명자원과학대학 동물생명자원과학과) ;
  • 조병욱 (부산대학교 생명자원과학대학 동물생명자원과학과)
  • Shin, Donghyun (Department of Animal Biotechnology, College of Agricultural and Life Sciences, Chonbuk National, University) ;
  • Kim, Kyoung Hwan (Department of Animal Science, College of Life Sciences, Pusan National University) ;
  • Lee, Ji Hyun (Department of Animal Science, College of Life Sciences, Pusan National University) ;
  • Cho, Byung-Wook (Department of Animal Science, College of Life Sciences, Pusan National University)
  • 투고 : 2018.05.28
  • 심사 : 2018.09.27
  • 발행 : 2018.10.30

초록

운동 후 분비되는 스트레스 호르몬인 cortisol을 통한 근육세포에 미치는 운동 스트레스의 재현과 coritisol 처리 농도에 따른 세포사멸, 세포질세망 스트레스 및 자가포식현상과의 관련성을 검증 하였다. 마우스 근육 세포주 C2C12를 배양하여 다양한 농도의 cortisol을 12시간 처리하여 세포의 형태 변화를 관찰하고, 세포사멸 마커인 IER3의 발현을 세포면역화학법을 이용하여 확인하였다. 또한 ER-stress와 자가포식 현상의 유도 여부를 확인하기 위하여 BiP와 LC3-I/LC3-II 항체를 이용하여 웨스턴 블랏법을 통해 검증 하였다. 그 결과 cortisol의 농도가 $50{\mu}g/ml$$100{\mu}g/ml$로 증가함에 따라 IER3와 BiP 및 LC3-II의 발현량도 유의적으로 증가함을 확인 할 수 있었다. 이러한 결과는 운동 스트레스 호르몬인 cortisol이 운동 후 근육세포의 세포사멸, 세포질세망 스트레스 및 자가포식에 영향을 미침을 보여준다. 본 연구결과는 호르몬과 근육세포 간의 관련성 연구에 기여할 것으로 기대된다.

Cortisol, a steroid hormone, functions within metabolism, immune response, and stress. Intense or prolonged physical exercise increases cortisol levels to enhance the gluconeogenesis pathway and stabilize blood glucose level. However, cortisol also exerts a negative impact on muscle function and creates a stressful environment in skeletal muscle cells. The present study investigated the function of cortisol as a stress hormone. To examine the effect of the exercise-induced hormone cortisol on skeletal muscles, C2C12 cells were cultured and treated with cortisol at different concentrations. As a result, we found that the morphology of C2C12 changed remarkably with 5 ug/ml cortisol treatment. Western blot analysis was conducted to learn whether ER-stress and autophagy were induced. We found that the expression ratio of LC3I/LC3II decreased and BiP expression increased after cortisol treatment. In addition, immunocytochemistry analysis with IER3 antibody clearly showed that apoptosis is induced after 12-hour cortisol treatment. These results indicate that cortisol treatment could induce apoptosis, ER-stress, and autophagy in muscle cells. This study would provide valuable information in the study of the effects of exercise on skeletal muscle cells and the development of additives to reduce cortisol stress.

키워드

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