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The role of collagenase and TIMP in Periodontal disease.

胶原酶和 TIMP 在牙周病中的作用。

基本信息

批准号：
02454443
负责人：
NOGUCHI Toshihide
金额：
$ 2.37万
依托单位：
Department of Periodontology, School of Dentistry, Aichigakuin University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1992
项目状态：
已结题

项目摘要

TIMP-1 concentrations in whole and gland saliva from healthy and periodontally diseased subjects were assayed by a sandwich enzyme immunoassay (sandwich EIA). TIMP-1 level in whole saliva of periodontally diseased subjects, 135 * 65 ng/ml(mean * SD), was clearly lower (p<0.01) than that of clinically healthy subjects, 273 * 145. Parotid (76 * 56 ng/ml) and submandibular-sublingual (124 70) saliva from healthy subjects showed significantly lower values than that in whole saliva. The TIMP-1 level in gingival crevicular fluid (GCF) from periodontal patients was found to be fairly low (31 * 24) compared to saliva samples. In the present study, collagenase activity was determined by applying a new strategy for the independent measurement of collagenase activity in TIMP-1-free samples prepared by the passage of the sample through an anti-TIMP-1 monoclonal antibody-affinity column. Both active {(1.21 * 0.63 units/ml (mean * SD)} and total (1.54 * 0.74) collagenase activities in TIMP-1-free wh … More ole saliva of diseased subjects were significantly higher than the activities (0.14 * 0.14 and 0.50 * 0.27) in TIMP-1-free whole saliva of healthy subjects. It is well known that various cytokines manifest potent proinflammatory and catabolic activities on matrix components and play key roles in periodontal tissue breakdown. The aim of this study was to investigate the effects of cytokines such as interleukin-1 (IL-1)alpha and beta, tumor necrosis factor -alpha(TNF-alpha), IL-6, IL-8 and transforming growth factor - beta1 (TGF-beta1) on the production of interstitial collagenase and tissue inhibitor of metalloproteinases-1 (TIMP-1) from human gingival fibroblasts (Gin-1). The results obtained were as follows: 1. The level of TIMP-1 in the culture media of Gin-1 stimulated by IL-1 alpha and beta and TNF-alpha was significantly increased in a dose-dependent manner. The level of TIMP-1 in the culture media stimulated by TGF-beta1 and IL-6 was increased slightly, but in the case of IL-8 it decreased. 2. Total collagenase activities in the culture media of Gin-1 stimulated by IL-1 alpha and beta, TNF-alpha and TGF-beta1 was significantly increased in a dose-dependent manner. These activities of IL-6 and IL-8 was increased slightly. 3. Active collagenase activities was significantly increased in the TIMP-1 free culture stimulated by IL-1 alpha and beta and TNF-alpha and it increased in a dose-dependent manner. These activities of TGF-beta1, IL-6 and IL-8 increased slightly. These findings indicate that some kinds of inflammatory cytokines may regulate the production of collagenase and it's inhibitor in the culture media of human gingival fibroblasts. Less

通过夹心酶免疫测定法（夹心EIA）测定健康和牙周病受试者的全唾液和腺体唾液中TIMP-1浓度。牙周病受试者的全唾液中TIMP-1水平为135 * 65 ng/ml（平均值 * SD），明显低于临床健康受试者的273 * 145 ng/ml（p<0.01）。腮腺（76 * 56 ng/ml）和下颌下舌下（124 * 70）唾液从健康受试者显示显着低于全唾液中的值。发现牙周病患者龈沟液（GCF）中的TIMP-1水平与唾液样品相比相当低（31 * 24）。在本研究中，胶原酶活性测定通过应用一种新的策略，通过抗TIMP-1单克隆抗体亲和柱的样品通过制备的TIMP-1-免费的样品中的胶原酶活性的独立测量。在无TIMP-1的wh中，活性胶原酶活性{（1.21 * 0.63单位/ml（平均值 * SD）}和总胶原酶活性（1.54 * 0.74）均高于对照组。 ...更多信息患病受试者的全唾液中的活性显着高于健康受试者不含TIMP-1的全唾液中的活性（0.14 * 0.14和0.50 * 0.27）。众所周知，各种细胞因子对基质成分表现出强有力的促炎和分解代谢活性，并在牙周组织破坏中发挥关键作用。本研究的目的是研究细胞因子如白细胞介素-1（IL-1）α和β、肿瘤坏死因子-α（TNF-α）、IL-6、IL-8和转化生长因子-β 1（TGF-β 1）对人牙龈成纤维细胞（Gin-1）产生间质胶原酶和金属蛋白酶组织抑制剂-1（TIMP-1）的影响。主要研究结果如下：1. IL-1 α、IL-1 β和TNF-α刺激的Gin-1细胞培养液中TIMP-1的水平显著升高，且呈剂量依赖性。在TGF-β 1和IL-6刺激的培养基中TIMP-1的水平略有增加，但在IL-8的情况下，TIMP-1的水平下降。2. IL-1 α和β、TNF-α和TGF-β 1刺激的Gin-1培养基中的总胶原酶活性以剂量依赖性方式显著增加。IL-6和IL-8的活性略有升高。3.在IL-1 α和β以及TNF-α刺激的无TIMP-1培养物中，活性胶原酶活性显著增加，并且以剂量依赖性方式增加。TGF-β 1、IL-6和IL-8的这些活性略有增加。这些结果表明，某些炎性细胞因子可能调节人牙龈成纤维细胞培养液中胶原酶及其抑制剂的产生。少