The role of collagenase and TIMP in Periodontal disease.

用双抗体夹心酶免疫分析法检测健康人和牙周病患者唾液和唾液中TIMP-1的浓度。牙周病患者全唾液中TIMP-1水平为135×65 ng/ml(Mean*SD)，明显低于临床健康者的273×145(p&lt；0.01)。健康人腮腺唾液(76×56 ng/ml)和颌下舌下唾液(124 70 ng/ml)明显低于全唾液。与唾液样本相比，牙周病患者牙周沟液中的TIMP-1水平相当低(31×24)。在本研究中，胶原酶活性是通过应用一种新的独立测量胶原酶活性的新策略来测定的，该胶原酶活性是通过抗TIMP-1单抗亲和层析柱制备的不含TIMP-1的样品制备的。在不含TIMP-1的Wh…中，胶原酶活性和总胶原酶活性均为(1.21×0.63单位/毫升(平均值*SD))和(1.54×0.74)疾病组唾液中的OL值明显高于健康组中不含TIMP-1的全唾液的活性(0.14±0.14和0.50±0.27)。众所周知，各种细胞因子在基质成分上表现出强烈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。本研究旨在探讨白介素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-β1(TGF-β1)等细胞因子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因子-1(TIMP-1)的影响。结果如下：1.IL-1α、β和TNF-α刺激的Gin-1细胞培养上清液中TIMP-1水平显著升高，并呈剂量依赖关系。在转化生长因子-β1和IL-6的刺激下，培养上清液中TIMP-1的水平略有升高，而在IL-8的刺激下，TIMP-1的水平下降。2.IL-1α和β、肿瘤坏死因子-α和转化生长因子-β1刺激的Gin-1细胞培养上清液中总胶原酶活性显著升高，且呈剂量依赖关系。IL-6、IL-8活性略有升高。3.在IL-1α、β-IL-1和肿瘤坏死因子-α刺激的无TIMP-1培养中，胶原酶活性显著升高，且呈剂量依赖性。这些转化生长因子-β1、IL-6和IL-8活性略有升高。提示某些炎性细胞因子可能对人牙龈成纤维细胞培养上清液中胶原酶及其抑制物的产生有调节作用。较少

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