华西口腔医学杂志

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机械应力对人牙周膜成纤维细胞整合素β1mRNA表达的调节

朱庆党1, 2, 巢永烈2, 陈新民2, 赵鹃1   

  1. 1.浙江大学医学院附属口腔医院修复科, 浙江杭州310006; 2.口腔疾病研究国家重点实验室, 四川大学, 四川成都610041
  • 收稿日期:2008-04-25 修回日期:2008-04-25 出版日期:2008-04-20 发布日期:2008-04-20
  • 通讯作者: 巢永烈,Tel:028- 85502335
  • 作者简介:朱庆党(1972-),男,云南人,主治医师,博士
  • 基金资助:

    国家自然科学基金资助项目(30700959);浙江省医药卫生科学研究基金资助项目(2007A131)

Regulation of integr in β1 mRNA expr ession by mechanical str ess in human per iodontal ligament fibroblasts

ZHU Qing- dang1,2, CHAO Yong- lie2,CHEN Xin- min2,ZHAO Juan1   

  1. 1. Dept. of Prosthodontics, The Affiliated Hospital of Stomatology, Medicine College of Zhejiang University, Hangzhou 310006, China; 2. State Key Laboratory of Oral Diseases, Sichuan University, Chengdu 610041, China
  • Received:2008-04-25 Revised:2008-04-25 Online:2008-04-20 Published:2008-04-20
  • Contact: CHAO Yong- lie,Tel:028- 85502335

摘要:

目的观察机械力刺激对人牙周膜成纤维细胞整合素β1 mRNA表达的影响。方法用Forcel四点弯曲加载装置通过对体外培养的人牙周膜成纤维细胞分别施加不同动态张、压应力(强度为1 000、2 000、4 000 μstrain,加力时间为0、0.5、1、4、8、12 h),采用实时荧光定量PCR法研究机械力对人牙周膜成纤维细胞整合素β1 mRNA表达的影响。结果施加动态的张、压应力后,人牙周膜成纤维细胞整合素β1 mRNA表达量下调,这种下调变化与所施加应力的性质、大小和作用持续时间相关。人牙周膜成纤维细胞整合素β1对张、压应力刺激的感应不完全一致,机械力刺激越强,整合素β1表达越明显。结论动态张、压应力刺激在本实验提供的微应力范围内可以促进目的基因mRNA表达改变,不同的机械力对细胞整合素β1效应不同。

关键词: 人牙周膜成纤维细胞, 张应力, 压应力, 整合素β1

Abstract:

Objective To investigate the effect of different kinds of mechanical stress on the mRNA expression of integrin β1 subunit in cultured human periodontal ligament fibroblasts(hPDLF). Methods To scalp and remove the periodontal ligament attached to the mid- third part of the fresh root of young premolars extracted for the cause of orthodontics. Cultured hPDLF by the method of digesting by Ⅰ- type collagenase combining with tissue adhering. Then hPDLF was isolated and purified by cells passage. The sixth passage′s cells were selected to be loaded. A new cyclic strain loading apparatus. Forcel four point bending device was used for mechanically loading. Cells were loaded by three levels(1 000, 2 000, 4 000 μstrain) of tensional and compressive forces and collected at different times(0, 0.5, 1, 4, 8, 12 h)course after strain loading. The quantity of integrin β1 mRNA in every group was analyzed by means of quantitative real- time PCR with the special primers of up- and down- regulated genes. Results Dynamic mechanical forces down- regulated the expression of integrin β1 subunit mRNA in hPDLF and the difference in groups by different magnitude, different kinds, and different time of mechanical forces loading were statistically significant. The stronger stimulated forces, the more down- regulated expression. Compression down- regulated the expression of integrin β1 subunit mRNA more than tension did. Conclusion Dynamic mechanical forces could regulate the expression of integrin β1 subunit mRNA. The difference among all the groups by different magnitudes, different kinds, and different time of mechanical forces loading were statistically significant.

Key words: human periodontal ligament fibroblasts, tensional force, compressive force, integrin β1