Journal of Veterinary Clinics (한국임상수의학회지)

The Korean Society of Veterinary Clinics (한국임상수의학회)
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Proteinases and their Inhibitors in Cartilage and Synovial Fluid Acquired from a Canine Osteoarthritic Model

개 퇴행성 관절염 모델을 이용한 연골과 활액 내 단백질 분해 효소와 억제제의 작용 연구

  • Seo, Jae-Won (Center for Healthcare Technology Development, Bio-Safety Research Institute, College of Veterinary Medicine, Chonbuk National University) ;
  • Lee, Hae-Beom (Center for Healthcare Technology Development, Bio-Safety Research Institute, College of Veterinary Medicine, Chonbuk National University) ;
  • Kim, Nam-Soo (Center for Healthcare Technology Development, Bio-Safety Research Institute, College of Veterinary Medicine, Chonbuk National University) ;
  • Lee, Young-Hoon (Institute of Oral Bioscience, School of Dentistry, Chonbuk National University) ;
  • Kang, Hyung-Sub (Center for Healthcare Technology Development, Bio-Safety Research Institute, College of Veterinary Medicine, Chonbuk National University) ;
  • Kim, In-Shik (Center for Healthcare Technology Development, Bio-Safety Research Institute, College of Veterinary Medicine, Chonbuk National University) ;
  • Park, Sang-Youel (Center for Healthcare Technology Development, Bio-Safety Research Institute, College of Veterinary Medicine, Chonbuk National University)
  • 설재원 (헬스케어 기술개발 사업단, 전북대학교 수의과대학 생체안전성 연구소) ;
  • 이해범 (헬스케어 기술개발 사업단, 전북대학교 수의과대학 생체안전성 연구소) ;
  • 김남수 (헬스케어 기술개발 사업단, 전북대학교 수의과대학 생체안전성 연구소) ;
  • 이영훈 (전북대학교 치과대학 구강생체과학연구소) ;
  • 강형섭 (헬스케어 기술개발 사업단, 전북대학교 수의과대학 생체안전성 연구소) ;
  • 김인식 (헬스케어 기술개발 사업단, 전북대학교 수의과대학 생체안전성 연구소) ;
  • 박상열 (헬스케어 기술개발 사업단, 전북대학교 수의과대학 생체안전성 연구소)
  • Published : 2009.04.30
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Abstract

Chondrocytes and synovial fluid derived markers are used to monitor for osteoarthritis(OA). Specific inhibitors, known as tissue inhibitors of metalloproteinases(TIMP), regulate the proteolytic activity of matrix metalloproteinases(MMP). This study investigated whether MMP and TIMP levels were altered in synovial fluid and cartilage following the experimental induction of OA in canines. Twenty mature beagle dogs underwent a unilateral surgical transection of the cranial cruciate ligament and the medial collateral ligament as well as a medial meniscectomy. Matrix metalloproteinase-2 and MMP-9 levels were assayed using Western blot and TIMP-2 levels were measured with enzyme-linked immunosorbent assays four weeks after OA induction. Increased MMP-2 expression was observed in chondrocytes isolated from cartilage following OA induction, but MMP-9 expression decreased. Matrix metalloproteinase-2 and MMP-9 levels in synovial fluid from the OA induced joint significantly increased compared to those of the sham group. Tissue inhibitors of metalloproteinase-2 concentrations were higher in chondrocytes from the OA cartilage, yet TIMP-2 remained lower in the synovial fluid of OA. This suggests the elevated release of MMP-9 over MMP-2 into the synovial fluid following the cartilage degradation-related death of chondrocytes after OA. Osteoarthritis can be further deteriorated by increased MMP activity in the synovial fluid because TIMP-2 exist low concentration into the extracellular matrix. As a result, MMP activity, particularly MMP-9 activity, can be useful as a biomarker in diagnosing and monitoring the early stages of canine OA.

퇴행성관절염(Osteoarthritis,OA)은 관절 부위의 퇴행성 변화가 특징이며, 이를 진단하기 위해서는 연골세포나 활액에서 유래된 표지인자가 일반적으로 사용된다. 이번 연구에서는 개를 이용하여 실험적으로 퇴행성관절염을 유도하고, 활액과 연골세포에서 단백질 분해 효소인 matrix metalloproteinase(MMPs)와 MMPs의 활성을 억제시키는 것으로 알려진 tissue inhibitors of metalloproteinases(TIMPs)의 발현 정도를 조사하였다. 20마리의 비글견이 퇴행성관절염 모델로 사용되었으며 MMP-2 와 -9은 Western blot 분석에 의해서, TIMP-2의 농도는 ELISA(enzyme-linked immunosorbent assays)에 의해 결정하였다. 퇴행성관절염 유도 4주 후에 연골에서 분리한 연골세포에서 MMP-2의 발현은 증가되었지만 MMP-9의 발현은 감소되었다. 그러나, 퇴행성관절염을 유도한 개의 활액에서는 MMP-2와 -9의 발현이 모두 증가하는 것을 보였다. TIMP-2의 농도는 퇴행성관절염을 유도한 연골에서 분리한 연골세포에서는 높았지만, 활액에서는 낮은 농도를 보였다. 이러한 결과는 MMP-9가 퇴행성관절염 시 연골 조직의 변성에 따른 연골세포의 손상에 의해 MMP-2보다 더 활액으로 방출된다는 것을 보여주며, 활액 내 TIMP-2의 감소에 따른 MMPs의 활성이 퇴행성관절염을 더욱 악화시키는 것을 제안해준다. 결국 MMPs의 활성은, 특히 MMP-9, 개의 퇴행성관절염의 조기 진단과 치료를 위한 표지인자로서 사용할 수 있을 것으로 사료된다.

Keywords

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