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MMP and TIMP production in periodontal ligament fibroblasts stimulated by Prevotella nigrescens lipopolysaccharide

손호현|이우철|김미리|양원경
대한치과보존학회 (Korean Academy Of Conservative Dentistry)|2005.09|Restorative Dentistry and Endodontics|제30권 제5호|pp. 372-384

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키워드

MMP-1수산화칼슘LPSTIMP-1MMP-2Ca(OH)2치주인대 섬유아세포P. nigrescensPDL fibroblast
주제분류치의학

초록

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-2 단백질 생성은 1, 10 ㎎/㎖ 에서 자극 시간과 비례하여 증가하였다. 3. TIMP-1 단백질 생성은 24시간까지 증가하다가 48시간에 감소하였고, 0.1과 1 ㎍/㎖ 에서 증가하였으나 10 ㎍/㎖ 에서 억제되었다. 4. P. nigrescens의 LPS를 수산화칼슘으로 처리시 MMP-1의 mRNA 발현은 현저하게 감소하였다.

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 ㎍/㎖) or LPS (10 ㎍/㎖) pretreated with 12.5 ㎎/㎖ 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 production 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 increase. 2. MMP-2 production time-dependently increased when stimulated with 1 and 10 ㎍/㎖ LPS, but there was no dose-dependent increase. 3. TIMP-1 production increased to 24 h, but decreased at 48 h. It increased when stimulated with 0.1 and 1 ㎍/㎖ LPS, but suppressed at 10 ㎍/㎖. 4. P. nigrescens LPS pretreated with Ca(OH)2 markedly downregulated MMP-1 gene expression.

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관련 논문AI추천

Prevotella nigrescens lipopolysaccharides로 자극된 MG63 세포에서 분비되는 기질금속단백질 MMP-1과 TIMP-1의 수준에 관한 연구김미리, 손원준, 손호현, 양원경, 엄정문, 이인복, 조병훈대한치과보존학회 · Restorative Dentistry and Endodontics2004 · 제29권 제5호Prevotella intermedia의 세균내독소가 치은섬유아세포와 치주인대세포에서의 matrix metalloproteinase 및 tissue inhibitor of metalloproteinase의 발현에 미치는 영향김성조, 김종관, 이주연, 최은영, 최인순, 최점일대한치주과학회 · Journal of Periodontal & Implant Science2005 · 제35권 제1호사람 치주인대섬유모세포에서 MMP-1, TIMP-1,2, Type 1 collagen, fibronectin 및 UNCL 발현에 미치는 H₂O₂와 chlorhexidine의 효과김병옥, 장현선, 최성미대한치주과학회 · Journal of Periodontal & Implant Science2008 · 제38권 제4호Prevotella intermedia 및 Prevotella nigrescens의 지질다당질이 대식 세포에서의 Interleukin-8 생성에 미치는 영향김성조대한치주과학회 · Journal of Periodontal & Implant Science2009 · 제39권 제2호사람 치은 섬유아세포에서의 Tannerella forsythia 전세균, 막단백질, 당지질에 의한 염증성 사이토카인 발현구영, 구기태, 김정은, 김태일, 류인철, 이성훈, 이용무, 정종평, 최봉규대한치주과학회 · Journal of Periodontal & Implant Science2008 · 제38권 제3호Correlation of expression and activity of matrix metalloproteinase-9 and -2 in human gingival cells of periodontitis patients김경아, 김철호, 노승현, 박영국, 송권호, 정수봉, 김한나현대한치주과학회 · Journal of Periodontal & Implant Science2013 · 제43권 제1호

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