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

虽然一些临床参数现在被用于牙周疾病的诊断，但对于应该使用哪些参数来评估疾病的进展速度、目前的疾病活动性和治疗的需要，仍然存在实际困难。人们普遍认为，对牙周液的分析可以提供特定代谢变化与疾病状态之间的关系的信息，并且牙周液中的各种蛋白水解酶在牙周病的发病机制中起着重要作用。本工作旨在阐明组织蛋白B、组织蛋白H、组织蛋白L、组织蛋白D、组织球蛋白G和髓质素等溶酶体蛋白在牙周组织病理破坏过程中的作用，并将这些酶在牙周疾病诊断中的应用价值。为此，我们制备了从人体来源纯化的各种酶的特异性抗体，并用酶免疫分析法检测了慢性成人牙周炎患者和实验性牙周炎患者牙周液中的酶水平。结果表明，半胱氨酸蛋白酶组织蛋白B、组织蛋白H和L在牙周炎较轻的阶段有选择性地释放到牙周间隙中，其水平与牙周炎的严重程度呈正相关。然而，在实验性牙周炎受试者的GCF中没有发现显著数量的这些酶。牙周炎患者牙周组织中丝氨酸蛋白酶、延髓蛋白和组织蛋白酶G的表达也随牙周炎的严重程度增加而增加，经牙周治疗后减少。有趣的是，在实验性牙周炎受试者的牙龈液中检测到大量的延髓素。结果表明，这些溶酶体蛋白参与了牙周炎的进展，牙周液中这些酶的动态变化有助于判断牙周炎的活动性和疾病进展。

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.