Journal of Nutrition and Health

The Korean Nutrition Society (한국영양학회)
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Effects of polygalacin D extracted from Platycodon grandiflorum on myoblast differentiation and muscle atrophy

길경에서 추출한 polygalacin D가 근원세포 분화 및 근위축에 미치는 영향

  • Eun-Ju Song (Department of Food and Nutrition, Sookmyung Women's University) ;
  • Ji-Won Heo (Department of Food and Nutrition, Sookmyung Women's University) ;
  • Jee Hee Jang (Department of Biological Sciences, Sookmyung Women's University) ;
  • Eonmi Kim (National Institute for Korean Medicine Development) ;
  • Yun Hee Jeong (National Institute for Korean Medicine Development) ;
  • Min Jung Kim (Department of Biological Sciences, Sookmyung Women's University) ;
  • Sung-Eun Kim (Department of Food and Nutrition, Sookmyung Women's University)
  • 송은주 (숙명여자대학교 식품영양학과) ;
  • 허지원 (숙명여자대학교 식품영양학과) ;
  • 장지희 (숙명여자대학교 생명시스템학부) ;
  • 김언미 (한국한의약진흥원 한약소재개발센터) ;
  • 정윤희 (한국한의약진흥원 한약소재개발센터) ;
  • 김민정 (숙명여자대학교 생명시스템학부) ;
  • 김성은 (숙명여자대학교 식품영양학과)
  • Received : 2023.10.12
  • Accepted : 2023.11.02
  • Published : 2023.12.31
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Abstract

Purpose: The balance between synthesis and degradation of proteins plays a critical role in the maintenance of skeletal muscle mass. Mitochondrial dysfunction has been closely associated with skeletal muscle atrophy caused by aging, cancer, and chemotherapy. Polygalacin D is a saponin derivative isolated from Platycodon grandiflorum (Jacq.) A. DC. This study aimed to investigate the effects of polygalacin D on myoblast differentiation and muscle atrophy in association with mitochondrial function in in vitro and in zebrafish models in vivo. Methods: C2C12 myoblasts were cultured in differentiation media containing different concentrations of polygalacin D, followed by the immunostaining of the myotubes with myosin heavy chain (MHC). The mRNA expression of markers related to myogenesis, muscle atrophy, and mitochondrial function was determined by real-time quantitative reverse transcription polymerase chain reaction. Wild type AB* zebrafish (Danio rerio) embryos were treated with 5-fluorouracil, leucovorin, and irinotecan (FOLFIRI) with or without polygalacin D, and immunostained to detect slow and fast types of muscle fibers. The Tg(Xla.Eef1a1:mitoEGFP) zebrafish expressing mitochondria-targeted green fluorescent protein was used to monitor mitochondrial morphology. Results: The exposure of C2C12 myotubes to 0.1 ng/mL of polygalacin D increased the formation of MHC-positive multinucleated myotubes (≥ 8 nuclei) compared with the control. Polygalacin D significantly increased the expression of MHC isoforms (Myh1, Myh2, Myh4, and Myh7) involved in myoblast differentiation while it decreased the expression of atrophic markers including muscle RING-finger protein-1 (MuRF1), mothers against decapentaplegic homolog (Smad)2, and Smad3. In addition, polygalacin D promoted peroxisome proliferator-activated receptor-gamma coactivator (Pgc1α) expression and reduced the level of mitochondrial fission regulators such as dynamin-1-like protein (Drp1) and mitochondrial fission 1 (Fis1). In a zebrafish model of FOLFIRI-induced muscle atrophy, polygalacin D improved not only mitochondrial dysfunction but also slow and fast muscle fiber atrophy. Conclusion: These results demonstrated that polygalacin D promotes myogenesis and alleviates chemotherapy-induced muscle atrophy by improving mitochondrial function. Thus, polygalacin D could be useful as nutrition support to prevent and ameliorate muscle wasting and weakness.

본 연구는 근생성 및 근위축 완화효능을 가진 유효소재 발굴의 필요성에 의해 polygalacin D가 근원세포 분화 및 미토콘드리아에 미치는 영향과 항암제 유도 근위축에 대한 완화효과를 각각 세포 및 동물실험을 통해 확인하고자 하였다. 그 결과, polygalacin D는 다핵을 지닌 근관세포의 수와 분화 종결인자인 MHC isoforms의 발현량을 증가시켰고 근육 내 단백질 분해 인자인 MuRF1, Smad2/3의 발현량은 유의적으로 감소시켰다. 또한 미토콘드리아 생합성 조절인자인 Pgc1α의 발현은 증가시키고 미토콘드리아 분열인자인 Drp1과 Fis1의 발현은 감소시켰다. 한편 zebrafish 동물모델을 통해 항암제 유도 근위축에 대한 개선효과를 확인한 결과, polygalacin D는 항암제에 의해 유도된 근위축과 미토콘드리아 손상을 완화시켰다. 이상의 결과들은 polygalacin D가 미토콘드리아 기능 증진을 매개로 근원세포 분화 촉진 및 근육 단백질 분해 저하 효과를 지닐 뿐만 아니라, 미토콘드리아 손상을 개선하여 항암제로 유도된 근위축에 대한 완화 효과를 나타냄을 시사한다. 따라서 본 연구를 통해 polygalacin D가 근생성 및 근위축 예방과 치료를 위한 잠재적인 유효소재로서의 가능성을 제시하였다.

Keywords

Acknowledgement

This work was supported by the National Research Foundation of Korea (NRF) funded by the Korea government (MSIT) (NRF-2020R1C1C1007553, 2023R1A2C1005313 to Sung-Eun Kim, NRF-2022R1F1A1074668 to Min Jung Kim).

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