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Prevotella nigrescens lipopolysaccharides로 자극된 MG63 세포에서 분비되는 기질금속단백질 MMP-1과 TIMP-1의 수준에 관한 연구

MMP-1 and TIMP-1 production in MG-63 cells stimulated with Prevotella nigrescens lipopolysaccharide

손원준|조병훈|이인복|손호현|김미리|엄정문|양원경
대한치과보존학회 (Korean Academy Of Conservative Dentistry)|2004.09|Restorative Dentistry and Endodontics|제29권 제5호|pp. 470-478

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참고문헌

주제분류치의학

초록

본 연구의 목적은 Prevotella nigrescens lipopolysaccharides (LPS)로 자극된 MG63 osteosarcoma 세포에서 생성, 분비되는 기질금속단백효소인 MMP-1과 그 억제제인 TIMP-1을 측정할 뿐아니라 수산화칼슘으로 처리한 P. nigrescens LPS에 의한 기질금속단백효소와 그 억제제의 분비수준의 변화를 알아보는데 있다. 혐기성 조건에서 배양한 P. nigrescens로부터 LPS를 추출하여 순수정제한 다음 0.1, 1 그리고 10 g/ml의 LPS 농도로 MG63 세포를 자극하거나 또는 수산화칼슘으로 처리한 10g/ml의 LPS로 세포를 다양한 자극하여 다양한 시간이 경과한 다음 세포로부터 분비되는 MMP-1과 TIMP-1의 RNA 수준을 real time-PCR 방법으로 측정하였다. 실험결과 MMP-1의 mRNA 수준은 48시간에서 최고에 달하였고 그 분비정도는 LPS의 농도에 비례하였다. TIMP-1 mRNA는 1g/ml의 세균성 LPS 자극시 24시간 및 48시간에서 높은 증가를 보였으나 고농도인 10g/ml의 LPS로 자극한 경우 오히려 그 발현이 억제되었다. 또한 수산화칼슘으로 전처리한 P. nigrescens LPS로 자극한 MG 63 세포에서는 MMP-1과 TIMP-1의 분비가 억제되었다. 이러한 결과를 통해 볼 때 P. nigrescens LPS에 의한 MMP-1과 TIMP-1의 발현조절이 치근단 질환에서 발생하는 치조골 흡수 기전중 하나로 사료된다. 뿐만 아니라 P. nigrescens에 의해 분비되는 기질금속단백효소를 매개로 하는 염증반응 감소에 수산화칼슘이 효과적으로 작용하는것으로 확인되어 치근단 질환에 관여하는 세균성 LPS를 제거하기 위해 임상적으로 사용되는 근거가 될 수 있다.

Abstract

The purpose of this study is to monitor the secretion of matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) produced by human osteosarcoma cell line (MG63) stimulated with Prevotella nigrescens lipopolysaccharides (LPS), and to compare the level of secretion before and after the treatment of calcium hydroxide on P. nigrescens LPS. LPS was extracted and purified from anaerobically cultured P. nigrescens. MG63 cells were stimulated by the LPS (0.1, 1, 10 ㎍/㎖) or LPS (10 ㎍/㎖) pretreated with 12.5 ㎎/㎖ of Ca(OH)2 for 3 days. Total RNA was isolated from the cell, and real-time quantitative polymerase chain reaction (PCR) was performed for quantification of MMP-1 and TIMP-1. The results were as follows.1. MMP-1 mRNA expression at 48 hr was highly increased by stimulation with P. nigrescens LPS. The increase was dose-dependent. 2. When stimulated with 1 ㎍/㎖ of LPS, TIMP-1 mRNA expression was highly increased at 24 hr and 48 hr. However, TIMP-1 expression was suppressed at higher concentration (10 ㎍/㎖). 3. When P. nigrescens LPS was pretreated with Ca(OH)2, MMP-1 and TIMP-1 gene expression was downregulated. The results of this study suggest that transcriptional regulation of MMP-1 and TIMP-1 by P. nigrescens LPS be one of the important mechanisms in bone resorption of periapical inflammation. The result of calcium hydroxide on MMP-1 and TIMP-1 gene expression suppression shows that calcium hydroxide detoxified bacterial LPS and thus should be used the medication of choice for intracanal dressings in root canal infected with black-pigmented bacteria.

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관련 논문AI추천

MMP and TIMP production in periodontal ligament fibroblasts stimulated by Prevotella nigrescens lipopolysaccharide김미리, 손호현, 양원경, 이우철대한치과보존학회 · Restorative Dentistry and Endodontics2005 · 제30권 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 제1호사람의 치수, 치은, 치주인대 세포에 tumor necrosis factor (TNF)-α로 자극 시 matrix metalloproteinase (MMPs)의 분비에 관한 연구김덕수, 김선영, 박상혁, 임은미, 최경규, 최기운대한치과보존학회 · Restorative Dentistry and Endodontics2011 · 제36권 제1호기계적 자극이 치주인대 세포의 osteoprotegerin과 receptor activator of nuclear factor κB ligand mRNA 발현에 미치는 영향옥승호, 이기주, 이승일, 전옥순, 황충주대한치과교정학회 · 대한치과교정학회지2005 · 제35권 제4호사람 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 제2호

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