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.

这项研究的目的是检查牙周机械应力引起的各种热休克蛋白的作用。将牙周韧带成纤维细胞（PDL）和MG-63成骨细胞样细胞接种到6孔培养板中，或涂有I型胶原蛋白的6孔柔性硅板。在汇合阶段，连续压缩应力的1,2,4 g/cmii D12或1秒拉伸/1秒弛豫，将最大的15％循环拉伸应力伸长延长了72小时。在刺激期结束时，将细胞匀浆进行分为10或15％的SDS聚酰胺凝胶电泳，并针对抗HSP 27、47、60、70抗体进行免疫印迹。连续压缩应力刺激了HSP 47、60的表达，但是HSP 70的产生在基线水平上几乎保持恒定，持续72小时。另一方面，在刺激12小时后，循环拉伸应力刺激了HSP 47、60和70的表达。 PDL表现出低ALPase活性的PDL倾向于产生HSP。在MG-63细胞中，抑制细胞增殖，并通过连续的压应激应诱导细胞死亡。使用TUNEL方法未检测到凋亡细胞。孵育12小时后，用压缩应力刺激了HSP27的产生，并且更大的压缩应力倾向于诱导更大座的HSP 27产生。 HSF-1的产生增加了12和24小时，但95 kDa频段的表达并没有明显变化。这些结果表明，HSP可能在正畸移动牙齿的牙周重建过程中起重要作用，但是HSP产生的模式可能会受到细胞类型和机械应力方式的影响。