Kennett C N, Cox S W, Eley B M
Department of Periodontology, King's College School of Medicine and Dentistry, London, UK.
Histochem J. 1997 Oct;29(10):727-34. doi: 10.1023/a:1026465118281.
Loss of tooth support during chronic periodontitis is very likely to involve tissue proteases such as cathepsin B. The distribution of this enzyme was, therefore, examined in ultrathin sections of gingival tissue embedded in acrylic resin and labelled with a sheep polyclonal antibody and gold-conjugated secondary antibody. Macrophages and fibroblasts in both inflamed and non-inflamed areas of tissue showed labelling, and this was strongest in lysosomes, corresponding to the normal intracellular location of cathepsin B. However, additional gold particles were found on the surface of these cells. Monocytes in inflamed areas also had surface labelling, some of which was present on microvilli. Labelled collagen fibres adjacent to all three cell types indicated that cathepsin B had been released into the immediate extracellular environment. Plasma membrane cathepsin B has previously been associated with cancers, but enzyme redistribution and release in the gingiva may have been linked to the inflammatory response, since fibroblasts and macrophages in non-inflamed areas showed less labelling of their surface and adjacent collagen. The collagen labelling added to evidence that cathepsin B can function extracellularly as well as intracellularly in connective tissue degradation. This destructive role for the enzyme is supported by our earlier measurements of increased biochemical activity in chronic periodontitis.
慢性牙周炎期间牙齿支持组织的丧失很可能涉及组织蛋白酶,如组织蛋白酶B。因此,用羊多克隆抗体和金标记的二抗对包埋于丙烯酸树脂中的牙龈组织超薄切片进行检测,以观察该酶的分布。组织炎症区和非炎症区的巨噬细胞和成纤维细胞均显示有标记,且在溶酶体中标记最强,这与组织蛋白酶B在细胞内的正常定位相符。然而,在这些细胞表面发现了额外的金颗粒。炎症区的单核细胞表面也有标记,其中一些存在于微绒毛上。与这三种细胞类型相邻的标记胶原纤维表明组织蛋白酶B已释放到紧邻的细胞外环境中。此前,细胞膜组织蛋白酶B与癌症有关,但牙龈中该酶的重新分布和释放可能与炎症反应有关,因为非炎症区的成纤维细胞和巨噬细胞表面及其相邻胶原的标记较少。胶原标记进一步证明组织蛋白酶B在细胞外以及细胞内均可参与结缔组织的降解。我们早期对慢性牙周炎中生化活性增加的测量结果支持了该酶的这种破坏作用。