Journal of Periodontal and Implant Science

Korean Academy of Periodontology (대한치주과학회)
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Involvement of apoptotic signals in cyclosporin A-induced proliferation of human gingival fibroblast

사람 치은 섬유모세포에서 Cyclosporin-A 유도 세포증식에 대한 항세포고사 기전

  • Jeong, Tea-Sul (Department of Periodontology, Chonnam National University) ;
  • Chung, Hyun-Ju (Department of Periodontology, Chonnam National University, Dental Science Research Institute, Chonnam National University) ;
  • Kim, Won-Jae (Department of Oral Physiology, Chonnam National University, Dental Science Research Institute, Chonnam National University)
  • 정태술 (전남대학교 치과대학 치주과학교실) ;
  • 정현주 (전남대학교 치과대학 치주과학교실, 전남대학교 치의학연구소) ;
  • 김원재 (전남대학교 구강생리학교실, 전남대학교 치의학연구소)
  • Published : 2005.09.30
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Abstract

Cyclosproin A(CsA)는 세포 이식거부방지를 위한 면역 억제제 및 자가 면역질환 치료제로 널리 사용되어 왔다. CsA는 매양된 사람 치은섬유아세포를 증식시킴이 알려져 있지만 CsA에 의한 세포증식기전에 대한 세포사멸기전 및 Bcl-2의 역할은 연구되어 있지 않다. 이번 연구는 사람 섬유아세포에서 CsA에 의한 세포증삭기전에 세포고사기전 및 Bcl-2 family가 관여하는지 밝히는 데에 목적이 있다. 세포 생장력은 MTT 방법으로 측정하였다. Bcl-2 family와 Fas 발현 정도는 RT-PCR 방법이나 western blot으로 확인하였다. Caspase-3 및 -9의 활성은 ELISER reader로, reactive oxygen species(ROS)는 fluorescence spectrometer에 의해 측정되었다. 미토콘드리아에서 세포질로 분비된 cytochrome c는 Western blot으로 조사하였다. CsA는 $0.1{\sim}10\;{\mu}M$에서 사람 섬유아세포의 생존률을 시간과 농도 의존적으로 증가시켰으며, 50 ${\mu}M$ CsA에서는 오히려 세포가 죽였다. 또한, CsA 처리로 미토콘드리아에서 세포질로 유리되는 cytochrome c 양과 VDAC 1 및 3 발현량이 감소되었고, caspase-9과 caspase-3의 활성도도 감소되었다. 한편, CsA 처리한 섬유아세포에서 death receptor 구성요소인 Fas 발현이 감소되었다. Bcl-2 family에 대한 RT-PCR, western blot 분석결과, 세포고사를 억제하는 Bel-2 발현은 증가되었으나 세포고사를 자극하는 Bax와 Bid의 발현은 감소되었다. 이러한 결과들은 사람 섬유아세포에서 CsA유도 세포증식에 Bcl-2 family와 ROS가 매개하는 미토콘드리아 의존 및 death receptor 의존 세포고사기전이 관여함을 시사하였다.

Keywords

References

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