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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.

本研究旨在探讨机械应激对人牙周膜成纤维细胞(PDL)热休克蛋白的诱导作用。PDL取自新鲜拔除的人前磨牙。从牙根中部1/3处取小块牙周组织，在含有10%致死牛血清的α-MEM培养液中孵育。传代培养长出的成纤维细胞样细胞，接种于包被I型胶原的6孔培养板或6孔柔性硅胶板中。在融合阶段，施加1，2，4，6g/cm2的连续压缩应力或1秒拉伸/1秒松弛，循环拉伸应力的最大伸长率为15%，作用时间为72小时。刺激期结束后，用PBS洗涤细胞，TCA固定，Urea-Triton-X溶液匀浆。细胞匀浆经10%或15%SDS-PAGE凝胶电泳，免疫印迹抗HSP27、47、60、70、90抗体。HSP47、60、70的表达在刺激20~30min后受到持续压应力的轻微刺激，并降至等于或低于0时间对照的强度。相反，HSP27的表达在120min内略有抑制，但几乎保持不变。HSP60和HSP70的表达在复苏后72h几乎不变。而HSP47、60和70的表达在刺激12h后受到周期性牵张应力的刺激。培养96h后，这些HSP的表达逐渐增强。提示HSP在正畸移动牙牙周组织中的表达受机械应力控制，在正畸移动牙牙周组织的重建过程中可能起重要作用。