Signal Transduction of Mechanical Stress by Heat Shock Proteins in Periodontal Ligaments Cells

牙周膜细胞中热激蛋白对机械应力的信号转导

基本信息

批准号：
09672091
负责人：
SHIMIZU Yoshiyuki
金额：
$ 1.92万
依托单位：
TOHOKU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09672091/
关键词：
Heat chock protein Periodontal ligament fibroblast Osteoblast Mechanical stress HSF-1 Orthodontic tooth movement Cell death HSP27 アポトーシスネクローシス HSP60 HSP70 HSP47

项目摘要

The purpose of this study was to examine the role of various heat shock proteins induced by mechanical stress in periodontium. Periodontal ligament fibroblasts (PDL) and MG-63 osteoblast-like cells were seeded into 6-well culture plates or 6-well flexible silicon plate coated with type I collagen. At the confluent phase, 1,2,4 g/cmィイD12ィエD1 of continuous compressive stress or 1 sec stretching/1 sec relaxation, maximum 15% elongation of cyclic stretching stress was applied for 72 hours. At the end of stimulation period, the cell homogenate was subjected into 10 or 15% SDS polyaclylamide gel electrophoresis and immunoblotted against anti-HSP 27, 47, 60, 70 antibody. Expressions of HSP 47, 60 were stimulated by continuous compressive stress, but HSP 70 production remained almost constant at baseline levels for 72 hours incubation. On the other hand, expression of HSP 47, 60 and 70 were stimulated by cyclic stretching stress after 12 hours of stimulation. PDL which showed low ALPase activity tended to produce HSPs remarkably. In MG-63 cells, cell proliferation was inhibited and the cell death was induced by continuous compressive stress. No apoptotic cells were detected by using TUNEL method. HSP27 production was stimulated with compressive stress after 12 hours of incubation and greater compressive stress tended to induce HSP 27 production in greater mount. HSF-1 production increased with compressive stress for 12 and 24 hours, but the expression of 95 kDa band didn't change markedly. These results indicate HSP may play an important role in the reconstruction process of a periodontium of orthodntically moving tooth, but the pattern of HSP production may be affected by the cell type and the mode of mechanical stress.

本研究的目的是检查牙周膜中机械应力诱导的各种热休克蛋白的作用，将牙周膜成纤维细胞 (PDL) 和 MG-63 成骨细胞样细胞接种到 6 孔培养板或 6 孔柔性硅片中。涂有 I 型胶原蛋白的板在汇合阶段，1,2,4 g/cm 的连续压缩应力或 1 秒拉伸/1 秒。松弛，施加最大15％的循环拉伸应力伸长72小时。在刺激期结束时，将细胞匀浆进行10或15％SDS聚丙烯酰胺凝胶电泳并针对抗HSP 27、47、60、70进行免疫印迹。 HSP 47、60 的表达受到持续压缩应力的刺激，但 HSP 70 的产生几乎保持在基线水平。另一方面，在12小时的刺激后，HSP 47、60和70的表达受到低ALPase活性的刺激，在MG-63细胞中倾向于产生不常见的HSP。通过TUNEL方法，在用压缩应力刺激HSP27产生后，没有检测到凋亡细胞。 12小时的孵育和更大的压缩应力往往会诱导更多的HSP 27产生，HSF-1的产生随着压缩应力12小时和24小时的增加而增加，但95 kDa条带的表达没有显着变化。这些结果表明HSP可能。 HSP在正畸移动牙齿牙周组织的重建过程中发挥着重要作用，但HSP的产生模式可能受到细胞类型和机械应力模式的影响。