Tuberculosis and Respiratory Diseases

  • 제57권1호
  • /
  • Pages.37-46
  • /
  • 2004
  • /
  • 1738-3536(pISSN)
  • /
  • 2005-6184(eISSN)
대한결핵및호흡기학회 (The Korean Academy of Tuberculosis and Respiratory Diseases)
권호 표지

허파혈관주위세포에서 저산소증에 의한 생존능의 억제와 유전자 발현의 변화

Inhibition of Viability and Genetic Change in Hypoxia-treated Lung Pericytes

  • 신종욱 (중앙대학교 의과대학 내과학 교실) ;
  • 김계영 (국립보건원 생명의약부 심혈관질환과) ;
  • 이영우 (중앙대학교 의과대학 내과학 교실) ;
  • 정재우 (중앙대학교 의과대학 내과학 교실) ;
  • 이병준 (중앙대학교 의과대학 내과학 교실) ;
  • 김재열 (중앙대학교 의과대학 내과학 교실) ;
  • 조인호 (국립보건원 생명의약부 심혈관질환과) ;
  • 박인원 (중앙대학교 의과대학 내과학 교실) ;
  • 최병휘 (중앙대학교 의과대학 내과학 교실)
  • Shin, Jong Wook (Department of Internal Medicine, Chung-Ang University School of Medicine) ;
  • Kim, Kae-Young (Division of Cardiovascular Diseases, Institute of Biomedical Research, Korean National Institute of Health) ;
  • Lee, Young Woo (Department of Internal Medicine, Chung-Ang University School of Medicine) ;
  • Jung, Jae Woo (Department of Internal Medicine, Chung-Ang University School of Medicine) ;
  • Lee, Byoung Jun (Department of Internal Medicine, Chung-Ang University School of Medicine) ;
  • Kim, Jae-Yeol (Department of Internal Medicine, Chung-Ang University School of Medicine) ;
  • Jo, Inho (Division of Cardiovascular Diseases, Institute of Biomedical Research, Korean National Institute of Health) ;
  • Park, In Won (Department of Internal Medicine, Chung-Ang University School of Medicine) ;
  • Choi, Byoung Whui (Department of Internal Medicine, Chung-Ang University School of Medicine)
  • 투고 : 2004.06.02
  • 심사 : 2004.07.06
  • 발행 : 2004.07.30
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초록

연구 배경 : 허파혈관주위세포는 허파미세혈관에서 혈액공기 장벽을 이루고 있는 중요한 세포이다. 이 세포는 생리학적으로 혈류와 혈관의 투과성을 조절하는 기능이 있다. 본 연구는 급성폐손상/급성호흡곤란증후군에서 혈관 과투과성 및 개형에 혈관주위세포의 변화가 중요한 역할을 할 것으로 보고 시작하게 되었다. 흰쥐로 부터 일차 배양한 허파혈관주위세포에 저산소 상태를 만들었을 때, 세포의 생존능에 미치는 영향과 저산소증에 의해 유도되는 유전자의 발현을 살펴보았다. 방 법 : 흰쥐로부터 허파혈관주위세포를 일차 배양 및 계대 배양하였다. 광학 현미경 및 세포 면역 화학 염색으로 세포를 확인하였다. 2% $O_2$의 세포 배양기와 $200{\mu}M$ $CoCl_2$를 처리하였다. 세포의 증식은 tryphan blue 염색 후 세포수를 세는 방법을 택하였다. 유전자 발현은 역전사 중합 효소 연쇄반응을 이용하였다. 결 과 : 1. 흰쥐로부터 허파혈관주위세포를 성공적으로 일차 배양 및 계대 배양할 수 있었다. 2. 2% $O_2$나 $CoCl_2$에서 혈관주위세포는 24시간, 36시간, 48시간에 증식이 억제되었다. 3. 허파혈관주위세포에 저산소 상태의 자극을 주면 VEGF와 smad-2의 발현이 현저하게 증가하였다. 4. 허파혈관주위세포의 HIF$1{\alpha}$, COX-2는 저산소상태에서 VEGF, smad-2에 비해 발현의 변화가 현저하지 않았다. 결 론 : 허파혈관주위세포를 일차 배양함으로써 허파꽈리혈관장벽의 연구를 위한 한 모델을 만들었고, 저산소 상태에서 증식의 억제와 유전자 발현의 변화를 살펴봄으로써 허파혈관손상의 기전을 규명하는데 향후 도움이 될 것으로 보인다.

Background : Lung pericytes are important constituent cells of blood-air barrier in pulmonary microvasculature. These cells take part in the control of vascular contractility and permeability. In this study, it was hypothesized that change of lung pericytes might be attributable to pathologic change in microvasculature in acute lung injury. The purpose of this study was how hypoxia change proliferation and genetic expression in lung pericytes. Methods : From the lungs of several Sprague-Dawley rats, performed the primary culture of lung pericytes and subculture. Characteristics of lung pericytes were confirmed with stellate shape in light microscopy and immunocytochemistry. 2% concentration of oxygen and $200{\mu}M$ $CoCl_2$ were treated to cells. Tryphan blue method and reverse transcription-polymerase chain reaction were done. Results : 1. We established methodology for primary culture of lung pericytes. 2. Hypoxia inhibited cellular proliferation in pericytes. 3. Hypoxia could markedly induce vascular endothelial growth factor(VEGF) and smad-2. 4. Hypoxia-inducible factor-$1{\alpha}$(HIF-$1{\alpha}$) was also induced by 2% oxygen. Conclusion : Viability of lung pericytes are inhibited by hypoxia. Hypoxia can stimulate expression of hypoxia-responsive genes. Pericytic change may be contributed to dysfunction of alveolar-capillary barrier in various pulmonary disorders.

키워드

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