Journal of Periodontal and Implant Science

Korean Academy of Periodontology (대한치주과학회)
권호 표지

The Effect of ${\Delta}^{12}PGJ2$ and $PPAR{\Gamma}$ Agonist on the Proliferation and Differentiation of Osteoblast

(${\Delta}^{12}PGJ_2$ 및 PPAR 감마 길항체가 조골세포의 증식 및 분화에 미치는 효과

  • Heo, Jeong-Mi (Department of Periodontology, College of Dentistry, Seoul National University) ;
  • Kim, Kyoung-Wha (Department of Periodontology, College of Dentistry, Seoul National University) ;
  • Chung, Kyoung-Wook (Department of Periodontology, College of Dentistry, Seoul National University) ;
  • Lee, Hye-Joon (Department of Periodontology, College of Dentistry, Seoul National University) ;
  • Rhyu, In-Chul (Department of Periodontology, College of Dentistry, Seoul National University) ;
  • Ku, Young (Department of Periodontology, College of Dentistry, Seoul National University, Intellectual Biointerface Engineering Center, KOSEF) ;
  • Chung, Chong-Pyoung (Department of Periodontology, College of Dentistry, Seoul National University, Intellectual Biointerface Engineering Center, KOSEF)
  • 허정미 (서울대학교 치과대학 치주과학교실) ;
  • 김경화 (서울대학교 치과대학 치주과학교실) ;
  • 정경욱 (서울대학교 치과대학 치주과학교실) ;
  • 이혜준 (서울대학교 치과대학 치주과학교실) ;
  • 류인철 (서울대학교 치과대학 치주과학교실) ;
  • 구영 (서울대학교 치과대학 치주과학교실, 지능형생체계면공학연구센터, 과학재단) ;
  • 정종평 (서울대학교 치과대학 치주과학교실, 지능형생체계면공학연구센터, 과학재단)
  • Published : 2005.03.30
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Abstract

1. 목적 Prostaglandin은 치주질환과 관련된 국소적 골 대사에 중요한 역할을 한다. ${\Delta}^{12}PGJ_2$는 생체 내에서 혈장의 존재 하에 형성되는 천연 $PGD_2$ 대사산물이며 peroxisome- proliferator에 의해 활성화되는 감마 수용체 (PPAR ${\Gamma}$)에 대해 높은 친화성을 갖는 리간드로서 핵 수용체군에 속하는 전사조절인자이다. 이 연구의 목적은 골화 과정에서 ${\Delta}^{12}PGJ_2$의 역할을 규명하기 위해, 조골세포주의 증식과 분화에 미치는 영향과 그에 관련된 세포기전을 조사하는 데에 있다. 2. 방법 인간 골육종세포주인 Saos-2 (ATCC.HTB 85)와 쥐의 조골세포주 (MC3T3-E1)를 배양한 후 실험군에 농도가 각각 $10^{-5}$, $10^{-6}$, $10^{-7}$, $10^{-8}$, $10^{-9}$ 몰인 ${\Delta}^{12}PGJ_2$와 ciglitazone (합성 PPAR 감마 길항체)를 첨가하였다. 조골세포에서 PPAR 감마의 발현을 관찰하기 위해 역전사효소-중합효소연쇄반응(RT-PCR)을 특정한 primer를 이용하여 시행하였다. 세포 증식은 1일, 2일, 3 일째에 MIT 분석법으로 측정하였고, 2 일째에 알칼리성 인산효소 (ALPase) 생산을 측정하였다. 위의 결과에서 얻은 적정한 농도에서 다양한 조골세포 분화의 표지자들-제 1 형 교원질, 알칼리성 인산효소, osteopontin 및 bone sialoprotein-에 대한 간이 정량적 역전사효소-중합효소연쇄반응 (semiquantitative RT-PCR)을 실시하였으며 골결절 형성에 대한 효과를 알아보고자 석회화 분석도 시행하였다. 3. 결과 ${\Delta}^{12}PGJ_2$와 ciglitazone 모두 Saos-2 세포주의 증식을 촉진시켰다 .$10^{-8}$ 몰의 ${\Delta}^{12}PGJ_2$와 $10^{-6}$몰의 ciglitazone을 첨가한 실험군을 대조군과 비교했을 때, 시간에 비례하여 세포 증식률이 증가되었다. 알칼리성 인산효소의 활성화 검사에서도 증식률에서와 유사한 결과를 보여주었다. 간이 정량적 RT-PCR에서는 ${\Delta}^{12}PGJ_2$로 처리한 군의 경우 제 1 형 교원질, 알칼리성 인산효소, osteopontin, 그리고 bone sialoprotein의 상대적 mRNA 수준이 유의하게 높았다. 석회화 분석에서는 MC3T3-E1 세포를 $10^{-6}$ 몰의 ${\Delta}^{12}PGJ_2$로 처리한 군과 $10^{-5}$ 몰의 ciglitazone으로 처리한 군에서 현저한 골결절 형성을 보였다. 이러한 결과들은 ${\Delta}^{12}PGJ_2$가 유용한 골 유도물질이 될 수 있으며 또한 그 작용기전이 PPAR 감마-의존형 경로와 연관되어 있음을 보여준다.

Keywords

References

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