Expressions and Functions of Heat Shock Proteins in Orthodontically Moving Tooth

正畸移动牙热休克蛋白的表达和功能

基本信息

批准号：
07672208
负责人：
SHIMIZU Yoshiyuki
金额：
$ 1.41万
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：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07672208/
关键词：
Heat shock protein Periodontal ligament fibroblasts Mechanical stress HSP 27 HSP 47 HSP 60 HSP 70 HSP 90 矯正的歯の移動骨芽細胞

项目摘要

The purpose of this study was to examine the induction of various heat shock proteins by mechanical stress in human periodontal ligament fibroblasts (PDL). PDL were obtained from freshly extracted human premolars. Small peaces of periodontium were taken from middle one-third of the root and incubated in alpha-MEM medium containing 10% fatal bovine serum. Outgrowing fibroblast-like cells were subcultured and seeded into 6-well culture plates or 6-well flexible silicon plate coated with type I collagen. At the confluent phase, 1,2,4,6g/cm^2 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 cells were washed with PBS and fixed with TCA.The cells were sclaped and homogenized in Urea-Triton-X solution. The cell homogenate was subjected into 10 or 15% SDS polyaclylamide gel electrophoresis and immunoblotted against anti-HSP 27,47,60,70,90 antibody. Expressions of HSP 47,60,70 were slightly stimulated by continuous compressive stress after 20-30 minutes of stimulation and were decreased to equal or lower intense of 0 time control. In contrast, expression of HSP 27 was slightly inhibited but almost constant during 120 minutes stimulation. Expressions of HSP60 and 70 were almost constant after recovery for 72 hours. On the other hand, expression of HSP 47,60 and 70 were stimulated by cyclic stretching stress after 12 hours of stimulation. Expressions of these HSP were increased to the end of incubation for 96 hours. These results indicated expressions of HSP were controlled by mechanical stress in PDL and HSP may play an important role in the reconstruction process of a periodontium of orthodontically moving tooth.

本研究的目的是检查机械应力对人牙周膜成纤维细胞（PDL）中各种热休克蛋白的诱导作用。 PDL 是从新鲜拔出的人前磨牙中获得的。从牙根的中间三分之一处取出一小块牙周组织，并在含有 10% 致命牛血清的 α-MEM 培养基中培养。将向外生长的成纤维细胞样细胞传代培养并接种到6孔培养板或涂有I型胶原的6孔柔性硅板中。在汇合阶段，施加1,2,4,6g/cm^2的连续压缩应力或1秒拉伸/1秒松弛，最大15%伸长的循环拉伸应力，持续72小时。刺激期结束时，用PBS洗涤细胞并用TCA固定。将细胞刮擦并在Urea-Triton-X溶液中匀浆。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、90抗体进行免疫印迹。刺激20-30分钟后，连续压缩应力轻微刺激HSP 47、60、70的表达，并降低至等于或低于0时间对照的强度。相反，HSP 27 的表达受到轻微抑制，但在 120 分钟刺激期间几乎保持恒定。恢复72小时后HSP60和70的表达几乎恒定。另一方面，刺激12小时后，循环拉伸应力刺激了HSP 47，60和70的表达。这些 HSP 的表达增加至孵育 96 小时结束。这些结果表明HSP的表达受PDL中机械应力的控制，HSP可能在正畸移动牙齿牙周组织的重建过程中发挥重要作用。