Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Adenoviral-Mediated Ref-1 Overexpression Potentiates NO Production in Bradykinin-Stimulated Endothelial Cells

Bradykinin으로 자극한 혈관내피세포에서 Ref-1의 세포내 과발현에 의한 NO 생성 증진 효과에 대한 연구

  • Song, Ju-Dong (Department of Microbiology & Immunology and Medical Research Institute, Pusan National University School of Medicine) ;
  • Kim, Kang-Mi (Department of Microbiology & Immunology and Medical Research Institute, Pusan National University School of Medicine) ;
  • Lee, Sang-Kwon (Department of Cardiovascular Surgery, Pusan National University) ;
  • Kim, Jong-Min (Department of Anatomy and Cell Biology, Dong-A University School of Medicine) ;
  • Park, Young-Chul (Department of Microbiology & Immunology and Medical Research Institute, Pusan National University School of Medicine)
  • 송주동 (부산대학교 의학전문대학원 미생물학 및 면역학교실) ;
  • 김강미 (부산대학교 의학전문대학원 미생물학 및 면역학교실) ;
  • 이상권 (부산대학교 흉부외과학교실) ;
  • 김종민 (동아대학교 의과대학 해부학교실) ;
  • 박영철 (부산대학교 의학전문대학원 미생물학 및 면역학교실)
  • Published : 2007.07.30
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Abstract

The dual-function protein redox factor-1 (Ref-1) is essential for base excision repair of oxidatively damaged DNA and also governs the activation of many redox-sensitive transcription factors. We examined the role of Ref-1 in regulation of nitric oxide (NO) synthesis employing adenoviral-mediatedoverexpression of Ref-1 in bradykinin-stimulated endothelial cells. Intracellular NO was detected with the NO-sensitive fluorophore DAF-2. Overexpression of Ref-1 potentiates bradykinin-stimulated NO production in endothelial cells. And, cells ifected with AdRef-1 showed higher fluorescence intensity compared with uninfected or AdD1312-infected cells. In parallel with this, over expression of Ref-1 also stimulated endothelial NO synthase (eNOS) enzyme activity, compared with unifected or AdD1312-infected cells, in bradykinin-stimulated cells as well as in unstimulated cells. These results suggest that Ref-1 implicates in endothelium-dependent vasorelaxation resulting from NO production in vascular system.

Redox Factor-1 (Ref-1)은 손상된 DNA의 복구 및 많은 세포내 산화환원에 민감한 transcription factors의 활성화에 기여하는 양면의 역할을 수행하는 단백질이다. 본 연구에서는 혈관내피세포에서의 nitric oxide (NO) 생성과정에서 Ref-1의 역할을 살펴보았다. Ref-1의 세포내 과발현을 위하여 adenoviral vector를 사용하였고 bradykinin으로 자극한 혈관내피세포에서 생성되는 NO 측정을 위하여 fluorophore DAF-2를 사용하였다. Ref-1 과 발현은 bradykinin으로 자극한 혈관내피세포의 NO 생성을 증가시켰다. 또한 자극되지 않은 Ref-1 과발현 세포는 viral vector로 감염되지 않은 그리고 control로 사용한 AdD1312로 감염된 세포보다 높은 fluorescence intensity를 나타내었다. 이와 비슷하게, Ref-1 과발현은 bradykinin으로 자극한 세포뿐만 아니라 자극하지 않은 세포에서도 감염되지 않은 그리고 AdD1312로 감염된 세포와 비교할 때 endothelial NO synthase (eNOS)의 활성을 크게 증가시켰다. 이는Ref-1 자신이 eNOS의 효소활성을 직접 조절할 수 있다는 것을 의미한다. 결론적으로 Ref-1이 혈관계에서 NO생성에 의해 기인되는 endothelium-dependent vasorelaxation에서 중요한 역할을 한다는 것을 시사한다.

Keywords

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