Screening System Establishment for Potential Anti-wrinkle Agents Using Human Fibroblast Elastase

엘라스타제를 이용한 주름개선 후보물질 검색 시스템의 구축

  • Oh, Mi-Hee (Department of Biochemistry, College of Medicine, Chung-Ang University) ;
  • Lee, Ju-Eun (Department of Biochemistry, College of Medicine, Chung-Ang University) ;
  • Kim, Su-Yeon (Department of Biochemistry, College of Medicine, Chung-Ang University) ;
  • Kim, So-Young (Department of Biochemistry, College of Medicine, Chung-Ang University) ;
  • Park, Kyoung-Chan (Department of Dermatology, Seoul National University College of Medicine) ;
  • Yun, Hye-Young (Department of Biochemistry, College of Medicine, Chung-Ang University) ;
  • Baek, Kwang-Jin (Department of Biochemistry, College of Medicine, Chung-Ang University) ;
  • Kwon, Nyoun-Soo (Department of Biochemistry, College of Medicine, Chung-Ang University) ;
  • Kim, Dong-Seok (Department of Biochemistry, College of Medicine, Chung-Ang University)
  • 오미희 (중앙대학교 의과대학 생화학교실) ;
  • 이주은 (중앙대학교 의과대학 생화학교실) ;
  • 김수연 (중앙대학교 의과대학 생화학교실) ;
  • 김소영 (중앙대학교 의과대학 생화학교실) ;
  • 박경찬 (서울대학교 의과대학 피부과학교실) ;
  • 윤혜영 (중앙대학교 의과대학 생화학교실) ;
  • 백광진 (중앙대학교 의과대학 생화학교실) ;
  • 권년수 (중앙대학교 의과대학 생화학교실) ;
  • 김동석 (중앙대학교 의과대학 생화학교실)
  • Published : 2009.03.30

Abstract

It has been reported that not only collagen but also elastin contribute to inhibit skin wrinkle formation. Ultraviolet (UV) radiation induces photo-aging on human skin. Because UV radiation increases elastase activity, it is thought that increased elastase activity could be the major reason for skin elasticity reduction and wrinkle formation by UV. In the present study to standardize elastase activity measuring system, purified elastases from porcine pancrease and human neutrophil, and cell extracts of normal human primary fibroblasts, 3T3 mouse fibroblasts, and CCD-25Sk human fibroblasts were used as various enzyme sources. Furthermore, elastase activities were compared according to concentrations of enzyme and substrate and incubation time. Phosphoramidon was used as a positive control to inhibit elastase activities of normal human primary fibroblasts and CCD-25Sk fibroblasts. However, it had no influence on the activity of porcine pancreatic elastase. Therefore, it is suggested that elastase used for testing anti-wrinkle agents should be selected carefully.

피부주름 개선에는 콜라겐(collagen)뿐만 아니라 탄력섬유인 엘라스틴(elastin) 등도 기여하는 것으로 보고되고 있다. 더 나아가 자외선에 의하여 사람의 피부에서 광노화 현상이 나타나며 자외선 조사 후 엘라스타제(elastase)의 활성이 증가하기 때문에 엘라스타제의 활성증가는 자외선에 의한 피부 탄성도의 감소 및 주름 생성의 주요원인으로 생각된다. 따라서 본 연구에서는 피부주름 생성에 영향을 미치는 엘라스타제의 활성을 측정하기 위한 모델을 마련하기 위하여 시판되는 두 가지 엘라스타제, 돼지 췌장 엘라스타제(porcine pancreatic elastase)와 사람 호중구 엘라스타제(human neutrophil elastase)를 사용하였으며 다음 세 가지는 정상 사람 섬유아세포(normal primary human fibroblasts), 쥐의 3T3 섬유아세포주(3T3 mouse fibroblasts), 사람의 CCD-25Sk 섬유아세포주(CCD-25Sk human fibroblasts)로부터 elastase를 준비하여 사용하였다. 준비된 5가지 효소의 농도에 기질의 농도 및 배양시간에 따르는 효소의 활성을 비교 평가하였다. 양성대조군으로 사용한 phosphoramidon이 normal human primary fibroblast elastase와 CCD-25Sk fibroblast elastase의 활성을 유의성 있게 억제한 반면에 porcine pancreatic elastase에는 별다른 영향을 미치지 못하였다. 따라서 주름 개선 후보물질 탐색을 위한 엘라스타제의 선정에는 신중한 고려가 필요할 것으로 사료된다.

Keywords

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