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Inhibitory Effects of Illicium verum Hooker fil. Dichloromethane Fractions on Adipocyte Differentiation

팔각회향 dichloromethane 분획물에 의한 지방세포 분화 억제 효과

  • Jeong, Hyun Young (Blue-Bio Industry regional Innovation Center, Dong-eui University) ;
  • Jeong, In Kyo (Department of Life Science and Biotechnology, Dong-eui University Graduate School) ;
  • Kim, Nam Ju (Department of Life Science and Biotechnology, Dong-eui University Graduate School) ;
  • Yun, Hee Jung (Department of Life Science and Biotechnology, Dong-eui University Graduate School) ;
  • Park, Jung Ha (Department of Life Science and Biotechnology, Dong-eui University Graduate School) ;
  • Kim, Byung Woo (Blue-Bio Industry regional Innovation Center, Dong-eui University) ;
  • Kwon, Hyun Ju (Blue-Bio Industry regional Innovation Center, Dong-eui University)
  • 정현영 (동의대학교 블루바이오 소재개발센터) ;
  • 정인교 (동의대학교 대학원 생명응용학과) ;
  • 김남주 (동의대학교 대학원 생명응용학과) ;
  • 윤희정 (동의대학교 대학원 생명응용학과) ;
  • 박정하 (동의대학교 대학원 생명응용학과) ;
  • 김병우 (동의대학교 블루바이오 소재개발센터) ;
  • 권현주 (동의대학교 블루바이오 소재개발센터)
  • Received : 2018.12.12
  • Accepted : 2019.04.11
  • Published : 2019.04.30

Abstract

Fat accumulation in adipocytes occurs through the process of adipogenesis in which preadipocytes differentiate into adipocytes. Obesity is a metabolic disorder caused by excessive accumulation of fat in the body, which increases the incidence of cardiovascular diseases, hypertension, type 2 diabetes, hyperlipidemia, and various cancers. Recently, inhibition of adipocyte differentiation was shown to be a potential antiobesity strategy. In this study, the inhibitory effect of dichloromethane fractions from Illicium verum Hooker fil. water extract on the differentiation of 3T3-L1 preadipocytes to adipocytes was investigated. Dichloromethane fractions from I. verum Hooker fil. significantly inhibited adipocyte differentiation when applied during the adipocyte differentiation process, as assessed by measuring fat accumulation using Oil-red O staining. In addition, dichloromethane fractions from I. verum Hooker fil. reduced important adipogenic transcription factors, such as CCAAT/enhancer binding protein ${\alpha}$ ($C/EBP{\alpha}$), $C/EBP{\beta}$, and peroxisome proliferator activated receptor ${\gamma}$ ($PPAR{\gamma}$). The expression of FAS and LPL, which are terminal differentiation markers of mature adipocytes, was also reduced in the 3T3-L1 adipocytes treated with dichloromethane fractions from I. verum Hooker fil. In addition, the treatment significantly inhibited mitotic clonal expansion, which is essential for adipocyte differentiation, by arresting the G1 phase of the cell cycle. Taken together, these results suggest that dichloromethane fractions from I. verum Hooker fil. may be a natural material with antiobesity effects.

비만은 에너지의 불균형으로 인하여 체내 지방조직에 지방이 축적되는 대사성질환으로 심혈관계 질환, 고혈압, 2형 당뇨, 고지혈증 및 각종 암의 발생 빈도를 증가시키는 요인이다. 지방의 축적은 지방전구세포가 지방세포로 분화하는 과정을 의미하는 adipogenesis라는 과정을 거쳐서 일어난다. 지방세포로의 분화는 다양한 호르몬과 전사인자들의 상호작용에 의해서 일어난다. 본 연구에서는 팔각회향이 항비만 소재로 활용 가능한지 확인하기 위해, 팔각회향 물 추출물을 분획하여 지방축적 억제 활성이 좋은 dichloromethane층을 선정하였다. 3T3-L1 지방전구세포가 성숙한 지방세포로 분화할 때 팔각회향 dichloromethane 층이 어떠한 기전으로 분화를 조절하는지 확인한 결과, 지방세포 분화에서 중요한 전사인자인 C/EBP family, $PPAR{\gamma}$의 발현이 억제되었고, 지방세포 최종 분화 마커로 알려져 있는 FAS 및 LPL의 발현 또한 감소되었다. 또한 G1기에서의 세포주기 정지를 통해 지방세포 분화 단계에서 필수적인 mitotic clonal expansion 단계를 억제한다는 결과를 얻었다. 이러한 연구 결과는 팔각회향이 항비만 효과를 가지는 천연물 소재로의 활용가능성을 보여주는 기초 자료가 될 것으로 사료된다.

Keywords

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Fig. 1. Extraction and fractionation of Illicium verum Hooker fil. by various organic solvents.

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Fig. 2. Effects of several fractions derived from lllicium verum Hooker fil. on lipid accumulation in 3T3-L1 adipocytes.

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Fig. 3. Effects of CH2Cl2 fraction derived from lllicium verum Hooker fil. on cell viability in 3T3-L1 pre-adipocytes.

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Fig. 4. Expression of adipogenesis-related proteins in 3T3-L1 cells treated with lllicium verum Hooker fil. CH2Cl2 fraction.

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Fig. 5. Anti-proliferative effects of lllicium verum Hooker fil. CH2Cl2 fraction in the mitotic clonal expansion process of adipocyte differentiation.

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Fig. 6. Cell cycle analysis of 3T3-L1 cells treated with of lllicium verum Hooker fil. CH2Cl2 fraction during the mitotic clonal expansion process of adipocyte differentiation.

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