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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.

本研究的目的是研究各种热休克蛋白在牙周组织中的作用，机械应力诱导。将牙周膜成纤维细胞（PDL）和MG-63成骨样细胞接种于6孔培养板或涂有I型胶原的6孔柔性硅胶板中。在汇合阶段，施加1, 2, 4g/cm × D12 × D1的连续压缩应力或1秒拉伸/1秒松弛，最大15%伸长率的循环拉伸应力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抗体进行免疫印迹。HSP 47、60的表达被持续的压应力刺激，但HSP 70的产生在72小时的孵育中几乎保持恒定在基线水平。另一方面，在刺激12小时后，通过周期性拉伸应力刺激HSP 47、60和70的表达。ALP活性低的PDL有明显的热休克蛋白生成倾向。在MG-63细胞中，持续压应力抑制细胞增殖并诱导细胞死亡。TUNEL法未检测到凋亡细胞。孵育12小时后，压缩应力刺激HSP 27产生，并且更大的压缩应力倾向于诱导更大量的HSP 27产生。HSF-1的表达随压力的增加而增加，但95 kDa条带的表达无明显变化。这些结果表明HSP可能在正畸移动牙牙周组织的重建过程中发挥重要作用，但HSP的产生模式可能受细胞类型和机械应力模式的影响。