THE DEVELOPMENT OF A NEW CLINICAL PARAMETER FOR DIAGNOSIS OF PERIODONTAL DISEASES AND THE NEW DRUG DESIGN

虽然现在有许多临床参数用于诊断牙周病，但存在实际困难的是，应该使用哪些参数来评估疾病的进展速度，目前的疾病活动和治疗的需要。龈沟液（Gingivalcrevicularfluid，GCF）的分析可以提供特定代谢变化与牙周病状态之间的关系，GCF中的多种蛋白水解酶在牙周病的发病机制中起着重要作用。本研究旨在阐明溶酶体蛋白酶包括组织蛋白酶B、H、L、D、G和胰蛋白酶在牙周组织病理破坏过程中的作用，并将这些酶的GCF水平的知识用于牙周疾病的诊断。为此，我们制备了从人类来源纯化的每种酶的特异性抗体，并通过酶免疫测定法测定了慢性成人牙周炎患者和实验性牙龈炎受试者GCF中酶的水平。结果表明，半胱氨酸蛋白酶组织蛋白酶B，H和L选择性地释放到龈沟在相对温和的牙周炎阶段和GCF中的水平与疾病的严重程度呈正相关。然而，在实验性牙龈炎受试者的GCF中没有发现显著量的这些酶。GCF中的丝氨酸蛋白酶Pegasin和组织蛋白酶G也显示出随着牙周炎的严重程度而增加，并通过牙周治疗而减少。有趣的是，在来自实验性牙龈炎受试者的GCF中检测到大量的GCF。结果表明，这些溶酶体蛋白酶参与牙周炎的进展，这些酶在GCF中的动力学可用于确定目前的疾病活动和疾病进展。

Although a number of clinical parameters are now used for diagnosis of periodontal diseases, there exists the practical difficulty as to which parameters should be used to assess the rate of progress of the disease, the present disease activity and the need for treatment. is generally accepted that analysis of gingival crevicular fluid (GCF) can provide information of association between specific metabolic change and disease status and that a variety of proteolytic enzymes in GCF play an important role in pathogenesis of periodontal disease. The present work was designed to clarify the roles of lysosomal proteinases including cathepsins B, H, L, D, G, and medullasin in pathological destructive process of periodontal tissues and to put the knowledge of the GCF levels of these enzymes to practical use for diagnosis of periodontal diseases. For this purpose, we prepared the specific antibodies to each enzyme purified from human sources and determined the levels of enzymes in GCF from chronic adult periodontitis patients and experimental gingivitis subjects by enzyme immunoassay. The results indicate that the cysteine proteinases cathepsins B, H, and L are selectively released into gingival crevices at a relatively mild stage of periodontitis and their levels in GCF are positively correlated with the severity of the disease. However, no significant amounts of these enzymes are found in GCF from experimental gingivitis subjects. The serine proteinases medullasin and cathepsin G in GCF are also shown to increase with the severity of periodontitis and to decrease by periodontal treatment. Interestingly, large amounts of medullasin was detected in GCF from experimental gingivitis subjects. The results indicate that these lysosomal proteinases participate in the progression of periodontitis and that the dynamics of these enzymes in GCF is useul for determining the present disease activity and the disease progression.