Restorative Dentistry and Endodontics

The Korean Academy of Conservative Dentistry (대한치과보존학회)
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MMP and TIMP production in periodontal ligament fibroblasts stimulated by Prevotella nigrescens lipopolysaccharide

Prevotella nigrescens lipopolysaccharide로 자극한 치주인대 섬유아세포에서 기질금속단백분해효소와 단백분해효소억제제의 생성 양상에 대한 연구

  • Yang, Won-Kyung (Department of Conservative Dentistry, College of Dentistry, Seoul National University) ;
  • Lee, Woo-Cheol (Department of Conservative Dentistry, College of Dentistry, Seoul National University) ;
  • Kim, Mi-Ri (Department of Conservative Dentistry, Dentistry, Asan Medical Center) ;
  • Son, Ho-Hyun (Department of Conservative Dentistry, College of Dentistry, Seoul National University)
  • 양원경 (서울대학교 치과대학 치과보존학교실) ;
  • 이우철 (서울대학교 치과대학 치과보존학교실) ;
  • 김미리 (서울아산병원 치과 보존과) ;
  • 손호현 (서울대학교 치과대학 치과보존학교실)
  • Published : 2005.09.01
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Abstract

The purpose of this study was to monitor the secretion of matrix metalloproteinase (MMP) and tissue inhibitor of metalloproteinase (TIMP) by human periodontal ligament (PDL) fibroblasts stimulated with Prevotella nigrescens lipopolysaccharide (LPS), and to examine the effect of calcium hydroxide treatment on P. nigrescens LPS. LPS was extracted and purified from anaerobically cultured P. nigrescens. PDL fibroblasts were stimulated by the LPS (0, 0.1, 1, 10 ${\mu}g/ml$) or LPS (10 ${\mu}g/ml$) pretreated with 12.5 mg/ml of $Ca(OH)_2$ for 3 days, for various periods of time (12, 24, 48 h). Immunoprecipitation were performed for protein level analysis of MMP-1 MMP-2 and TIMP-1. Total RNA was isolated and real-time quantitative polymerase chain reaction (PCR) was performed for quantification of MMP-1 mRNA. According to this study, the results were as follows: 1. The p개duction of MMP-1 by stimulation with P. nigrescens LPS increased in time-dependent manner, and showed maximum value at 48 h in both protein and mRNA level. But there was no dose-dependent increas. 2. MMP-2 production time-dependently increased when stimulated with 1 and 10 ${\mu}g/ml$LPS, but there was no dose-dependent increase. 3. TIMP-1 p개duction increased to 24 h, but decreased at 48 h. It increased when stimulated with 0.1 and 1${\mu}g/ml$, but suppressed at 10 ${\mu}g/ml$ .4. P. nigrescens LPS pretreated with $Ca(OH)_2$ markedly downregulated MMP-1 gene expression.

이 연구에서는 Prevotella nigrescens (P. nigrescens)의 lipopolysaccharide (LPS)로 자극한 치주인대 섬유아세포에서 matrix metalloproteinase (MMP)와 tissue inhibitor of metalloproteinase (TIMP)의 생성 양상과, LPS를 수산화칼슘으로 처리했을 때의 영향을 평가하였다. P. nigrescens에서 추출, 정제한 여러 농도의 LPS와 수산화칼슘으로 처리한 LPS로 치주인대 섬유아세포를 자극하여, Immunoprecipitation법으로 MMP-1, -2, TIMP-1의 단백질 생성 양상을, real-time polymerase chain reaction법으로 MMP-1의 mRNA 발현 양상을 분석하였다. 이 연구의 결과는 아래와 같다. 1. MMP-1은 단백질과 유전자 수준 모두 자극 시간과 비례하여 증가하여 48시간에 최대값을 보였다. 2. MMP-2단백질 생성은 1, 10 mg/ml에서 자극 시간과 비례하여 증가하였다. 3. TIMP-1 단백질 생성은 24시간까지 증가하다가 48시간에 감소하였고, 0.1과 1 ${\mg}g/ml$에서 증가하였으나 10 ${\mu}g/ml$ 에서 억제되었다. 4. P. nigrescens의 LPS를 수산화칼슘으로 처리시 MMP-1의 mRNA 발현은 현저하게 감소하였다.

Keywords

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