Korean Journal of Clinical Laboratory Science (대한임상검사과학회지)

Korean Society for Clinical Laboratory Science (대한임상검사과학회)
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Role of p-anisaldehyde in the Differentiation of C2C12 Myoblasts

C2C12 근육모세포의 분화에서 p-anisaldehyde의 역할

  • Dal-Ah KIM (Ewha Medical Research Center, Ewha Womans University College of Medicine) ;
  • Kyoung Hye KONG (Ewha Medical Research Center, Ewha Womans University College of Medicine) ;
  • Hyun-Jeong CHO (Department of Biomedical Laboratory Science, College of Medical Science, Konyang University) ;
  • Mi-Ran LEE (Department of Biomedical Laboratory Science, College of Health Science, Jungwon University)
  • 김달아 (이화여자대학교 의과대학 의과학연구소) ;
  • 공경혜 (이화여자대학교 의과대학 의과학연구소) ;
  • 조현정 (건양대학교 의과학대학 임상병리학과) ;
  • 이미란 (중원대학교 의료보건대학 임상병리학과)
  • Received : 2023.08.04
  • Accepted : 2023.08.25
  • Published : 2023.09.30
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Abstract

In this study, we investigated whether p-anisaldehyde (PAA), the main component of essential oils derived from anise seeds, influences the differentiation of mouse C2C12 myoblasts. Cells were induced to differentiate over 5 days using a differentiation medium with or without PAA (50 or 200 mg/mL). Myotube length and diameter were measured, and the expressions of myogenic markers (myoblast determination protein 1, myogenin, myocyte enhancer factor 2, muscle creatine kinase, and myosin heavy chain) and atrophy-related genes (atrogin-1 and muscle ring finger-1 [MuRF-1]) were assessed by quantitative real-time polymerase chain reaction. Additionally, protein kinase B (Akt) phosphorylation was monitored by western blotting. PAA significantly induced the formation of smaller and thinner myotubes and reduced myogenic marker expression. Furthermore, PAA increased the expressions of atrogin-1 and MuRF-1 and simultaneously reduced Akt phosphorylation. Our findings indicate that PAA inhibits the myogenic differentiation of C2C12 cells by reducing the phosphorylation and activation of Akt.

골격근은 대사, 열기반 온도 조절, 그리고 전반적인 체내 균형을 위해 필수적인 조직이고 근발생(myogenesis)이라는 다단계 과정을 거쳐서 근관세포를 형성한다. p-아니스알데하이드(p-anisaldehyde, PAA) (4-메톡시벤잘데하이드)는 아니스 씨에서 추출된 에센셜 오일의 주성분이지만, 골격근에서의 기능은 아직까지 알려져 있지 않다. 따라서, 우리는 마우스 C2C12 근육모세포를 이용하여 근육분화가 PAA에 의해 영향을 받는지를 연구하였다. C2C12 근육모세포의 분화를 유도하기 위해 이 세포를 분화배지에서 5일동안 배양하였고, 매일 PAA (50 또는 200 ㎍/mL)를 포함하는 새로운 배지로 교체하였다. 대조군으로서 PAA가 포함되지 않은 배지를 사용하였다. 우리는 분화시작 후 1, 3, 5일째에 근관세포의 길이와 지름을 측정함으로써 PAA가 근관 형성에 미치는 영향을 평가하였고, quantitative real-time polymerase chain reaction 분석을 통해 PAA가 근육 표지인자(myoblast determination protein 1, myogenin, myocyte enhancer factor 2C, muscle creatine kinase, 및 myosin heavy chain)와 근육위축 관련 유전자(atrogin-1과 muscle ring finger-1 [MuRF-1])의 발현에 미치는 영향을 분석하였다. 또한, 주요 근육형성 키나아제인 protein kinase B (Akt)의 인산화를 웨스턴 블롯을 이용해 관찰하였다. 그 결과 PAA가 더 작고 얇은 근관 형성을 유의하게 유발하며 근육 표지인자의 발현을 감소시킨다는 것을 확인하였다. 또한, atrogin-1과 MuRF-1의 발현이 PAA에 의해서 감소하였는데, 이는 Akt 인산화의 감소와 일치하는 결과이다. 결론적으로, 본 연구결과는 PAA가 Akt 인산화와 활성화를 감소시킴으로써 C2C12 세포에서의 근육 분화를 억제하는 역할을 한다는 것을 증명한다.

Keywords

Acknowledgement

This research was supported by "Regional Innovation Strategy (RIS)" through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (MOE) (2021RIS-001(1345370811)), and Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (2021R1I1A1A01041710) (2019R1I1A1A01055061).

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