A study on possible roles of granulocyte neutral proteinases and the control mechanism of their activities in the process of periodontal breakdown

粒细胞中性蛋白酶在牙周破坏过程中的可能作用及其活性控制机制的研究

• 批准号: 05671600
• 负责人: KUNIMATSU Kazushi
• 金额: $ 1.34万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05671600/
• 关键词: cathepsin G; medullasin; periodontal disease; lysosomal enzyme; experimental gingivitis; protease inhibitor; gingival crevicular fluid; 実験的歯肉炎; カテプシンE; カテプシンD; 好中球; 炎症歯肉

To clarify the role of host-derived lysosomal proteinases in the mechanism of periodontal breakdown, cathepsin G and medullasin, two major serine proteinases, were investigated. In gingival crevicular fluid (GCF) obtained from periodontitis patients, almost all activities of both enzymes were not detected, while the amounts of both enzymes determined by enzyme-linked immunosorbent assay increased with increasing GCF volume, indicating the participation of the enzymes in the progression of periodontitis. The reason why their activities were lost in GCF is suggested that they were blocked by their specific serum-derived proteinase inhibitor, especially alpha_1-proteinase inhibitor, since immunoblotting analyzes of GCF using each enzyme and alpha_1-proteinase inhibitor revealed a weak protein band of the respective enzyme and the inhibitor alone and a strong band of the complex of those. In experimental gingivitis of volunteers induced by refraining from all oral hygiene measures for 21 d … More ays, a remarkable increase of these enzymes in GCF were shown prior to the detection of inflammation monitored by various clinical parameters, suggesting the involvement in the development of inflammation. In inflamed gingival tissues, these enzymes were restricted in neutrophils and macrophages and concentrated especially in the vicinity of pocket epithelium partly with degranulation extracelluarly. This finding suggests the participation of these enzymes in direct degradation of host tissues besides the host-defensive mechanism against bacterial pathogens. Moreover, purified enzymes injected into rabbit gingivae caused inflammatory response characterized by noticeable injury to vessels and mild infiltration of inflammatory cells including neutrophils and macrophages. The function of the enzymes, however, were completely lost with the treatment by their specific inhibitor, alpha_1-proteinase inhibitor or alpha_1-antichymotrypsin. From the findings in this study, cathepsin G and medullasin are present in GCF and gingival tissue and participated in the host-defensive mechanism modulated by serum-derived proteinase inhibitors. Once the enzyme-inhibitor balance is lost, these two enzymes conversely play a destructive role against periodontal tissues. Less

为了阐明宿主来源的溶酶体蛋白酶在牙周破坏机制中的作用，研究了两种主要的丝氨酸蛋白酶组织蛋白酶G和丝氨酸蛋白酶。在龈沟液（GCF）从牙周炎患者，几乎所有的活动，这两种酶都没有检测到，而酶联免疫吸附测定测定的两种酶的量增加，随着GCF体积的增加，表明参与牙周炎的进展中的酶。它们在GCF中失去活性的原因表明它们被它们特异性的血清衍生的蛋白酶抑制剂，特别是α_1-蛋白酶抑制剂阻断，因为使用每种酶和α_1-蛋白酶抑制剂的GCF的免疫印迹分析揭示了各自的酶和单独的抑制剂的弱蛋白条带和它们的复合物的强条带。在实验性牙龈炎的志愿者避免所有口腔卫生措施21天 ...更多信息 总之，在通过各种临床参数监测的炎症检测之前，GCF中这些酶显着增加，表明参与炎症的发展。在发炎的牙龈组织中，这些酶仅限于中性粒细胞和巨噬细胞，尤其是在袋上皮附近集中，部分伴有细胞外脱颗粒。这一发现表明，这些酶参与宿主组织的直接降解，除了对细菌病原体的宿主防御机制。此外，纯化的酶注射到兔牙龈引起炎症反应，其特征在于明显的血管损伤和炎症细胞，包括中性粒细胞和巨噬细胞的轻度浸润。但用α_1-蛋白酶抑制剂或α_1-抗糜蛋白酶抑制剂处理后，这些酶的功能完全丧失。从这项研究的结果来看，组织蛋白酶G和牙龈蛋白酶存在于龈沟液和牙龈组织中，并参与了由血清来源的蛋白酶抑制剂调节的宿主防御机制。一旦酶-抑制剂平衡失去，这两种酶反过来对牙周组织起破坏作用。少

项目成果

Kunimatsu K.et al: "Distribution of cathepsins E and D in inflamed gingival tissues." Journal of Dental Research. 73. 901 (1994)

Kunimatsu K.et al：“组织蛋白酶 E 和 D 在发炎牙龈组织中的分布。”

Kunimatsu K.et al: "Immunohistochemical study of cathepsin G and medullasin in inflamed gingivae." Journal of Dental Research. 73. 906 (1994)

Kunimatsu K.et al：“组织蛋白酶 G 和髓质素在发炎牙龈中的免疫组织化学研究。”

Kunimatsu K., Mine N., Muraoka Y., Kato I., Hase T., Aoki Y., Yamamoto K.: "Identification and possible function of cathepsin G in gingival crevicular fluid from chronic adult periodontitis patients and from experimental gingivitis subjects" Journal of Pe

Kunimatsu K.、Mine N.、Muraoka Y.、Kato I.、Hase T.、Aoki Y.、Yamamoto K.：“慢性成人牙周炎患者和实验性牙龈炎受试者龈沟液中组织蛋白酶 G 的鉴定和可能功能

Kunimatsu K.et al.: "Identification and possible function of cathepsin G in gingival crevicular fluid from chronic adult periodontitis patients and from experimental gingivitis subjects." Journal of periodontal Research. 30. 51-57 (1995)

Kunimatsu K.等人：“慢性成人牙周炎患者和实验性牙龈炎受试者龈沟液中组织蛋白酶 G 的鉴定及其可能的功能。”

Kunimatsu K.et al.: "Distributuin of cathepsins E and D in inflamed gingival tissues." Journal of Dental Research. 73. 901- (1994)

Kunimatsu K.et al.：“组织蛋白酶 E 和 D 在发炎牙龈组织中的分布。”