Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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A Study on the Gene Expression in Shikonin-Induced Inhibition of Adipogenesis

Shikonin에 의한 지방세포형성 억제과정에서의 유전자 발현 연구

  • Lee, Hae-Yong (Department of Microbiology, Chung-Ang University College of Medicine) ;
  • Kang, Ryun-Hwa (Department of Microbiology, Chung-Ang University College of Medicine) ;
  • Chung, Sang-In (Department of Microbiology, Chung-Ang University College of Medicine) ;
  • Cho, Soo-Hyun (Department of Family Medicine, Yongsan Hospital, Chung-Ang University) ;
  • Oh, Dong-Jin (Department of Internal Medicine, Chung-Ang University College of Medicine) ;
  • Yoon, Yoo-Sik (Department of Microbiology, Chung-Ang University College of Medicine)
  • 이해용 (중앙대학교 의과대학 미생물학교실) ;
  • 강련화 (중앙대학교 의과대학 미생물학교실) ;
  • 정상인 (중앙대학교 의과대학 미생물학교실) ;
  • 조수현 (중앙대학교 용산병원 가정의학과) ;
  • 오동진 (중앙대학교 의과대학 내과학교실) ;
  • 윤유식 (중앙대학교 의과대학 미생물학교실)
  • Published : 2009.11.30
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Abstract

Shikonin, a component of Lithospermum erythrorhizon Sieb. et Zucc, exerts various characteristics such as anti-inflammatory, anti-cancer and anti-obesity functions. To elucidate the molecular mechanism of shikonin-induced inhibition of adipogenesis, we analyzed the mRNA expression level of various adipogenesis-related factors including C/EBPs (CCAAT/enhancerbinding proteins) and $PPAR{\gamma}$ (peroxisome proliferator-activated receptor $\gamma$). The data showed that mRNA expressions of C/$EBP{\beta}$ and C/$EPB{\delta}$ were only slightly changed by shikonin treatment, but mRNA expressions of $PPAR{\gamma}$ and C/$EPB{\alpha}$ were significantly down-regulated. Then, we tested whether upstream regulators of C/$EBP{\beta}$ and $PPAR{\gamma}$ were involved in anti-adipogenesis of shikonin. C/$EBP{\gamma}$ and CHOP (C/EBP homologous protein), which are upstream regulators of C/$EBP{\beta}$, were not affected by shikonin treatment. On the contrary, the mRNA level of KROX20 was markedly down-regulated by shikonin treatment. These results suggest that KROX20 might regulate downstream factors of adipogenesis through C/$EBP{\beta}$-independent pathway. The expression of KLF15 (Kruppel-like factor15), which is a member of KLF family and is a upstream regulator of $PPAR{\gamma}$, was dramatically decreased by shikonin treatment, but KLF2 was not changed. Shikonin had no impact on the expression of KLF5 in the early stage of adipogenesis, but shikonin increased expression of KLF5 in the late stage of adipogenesis. Even though mRNA expression of KLF5 was moderately changed by shikonin treatment, its effect may be small compared with the effect of KLF15, which was markedly inhibited. Taken together, these results suggest that shikonin inhibits adipogenesis through the down-regulation of $PPAR{\gamma}$ and C/$EPB{\alpha}$, which is mediated by the down-regulation of two pro-adipogenic factors, KROX20 and KLF15.

천연생약 자초의 한 성분인 shikonin은 항염증, 항암 및 항비만 등 다양한 분야에 효과를 보여왔다. 이번 연구에서는 shikonin이유도하는 adipogenesis억제 과정에 어떤 인자들이 작용하는지 살펴보았다. Shikonin의 효과에 대한 분자적 메커니즘을 규명하기 위해, real-time PCR을 이용하여 C/EBPs, $PPAR{\gamma}$를 포함한 다양한 adipogenesis 인자들의 mRNA 발현량을 분석하였다. 그 결과, 초기 분화의 주요 조절자인 C/$EBP{\beta}$와 C/$EPB{\delta}$는 shikonin에 의해 거의 변화가 없었으나, 후기 분화의 주요 조절자인 $PPAR{\gamma}$와 C/$EPB{\alpha}$의 mRNA 발현은 유의하게 감소하였다. Shikonin에 의한 adipogenesis억제의 메커니즘을 좀 더 자세히 밝히기 위해 adipogenesis과정의 상위 단계에 위치한 조절자들의 mRNA 발현을 분석하였다. C/$EBP{\beta}$의 상위 조절자인 C/$EBP{\gamma}$, CHOP은 shikonin에 의해 영향을 받지 않았으나, KROX20의 경우 유의하게 감소하였다. 이러한 결과는 Pro-adipogenic 인자인 KROX20의 감소가 C/$EBP{\beta}$에 영향을 주기 보다는 C/$EBP{\beta}$와 독립적으로 그 하위의 인자들에게 영향을 줄 수 있음을 제시한다. $PPAR{\gamma}$의 상위 조절자로 알려져 있는 KLF 들 중에서 pro-adipogenic 인자인 KLF15의 mRNA 발현은 shikonin에 의해 급격히 감소하였으나 anti-adipogenic 인자인 KLF2는 shikonin에 의한 변화가 거의 없었다. 또 다른 pro-adipogenic 인자인 KLF5의 경우, 주로 작용하는 초기 분화에서는 shikonin에 의해 거의 변화가 없었지만, 후기 분화에서는 조금 증가하였다. 이러한 후기 분화에서의 KLF5의 변화는 KLF15에 비해 전체 분화에 크게 영향을 주지 못하는 것 같다. 결론적으로, shikonin은 pro-adipogenic 인자인, KROX20과 KLF15의 조절을 통해 $PPAR{\gamma}$ 및 C/$EPB{\alpha}$의 mRNA 발현을 억제함으로써 지방 세포의 분화를 저해한다고 사료된다.

Keywords

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