华西口腔医学杂志 ›› 2012, Vol. 30 ›› Issue (5): 463-467.doi: 10.3969/j.issn.1000-1182.2012.05.004

• 基础研究 • 上一篇    下一篇

张、压应力刺激下人牙周膜成纤维细胞早期增殖活性和差异表达基因的变化

范晓枫1 王羽2,3 李宇4,5 赵志河4,5   

  1. 1.海南口腔医院正畸科, 海口570125; 2.中山大学光华口腔医学院·附属口腔医院正畸科;3.广东省口腔医学重点实验室, 广州510055; 4.四川大学华西口腔医院正畸科;5.口腔疾病研究国家重点实验室, 四川大学, 成都610041
  • 收稿日期:2011-03-11 修回日期:2011-09-10 出版日期:2012-10-01 发布日期:2012-10-01
  • 通讯作者: 赵志河,Tel:028-85503645
  • 作者简介:范晓枫(1974—),女,河南人,主治医师,博士
  • 基金资助:

    国家自然科学基金资助项目(10772128);高等学校博士学科点专项科研基金资助项目(20060610083)

Early proliferation changes and differences of gene expression in human periodontal ligament fibroblasts subjected to tensile and compressive stress

Fan Xiaofeng1, Wang Yu2,3, Li Yu4,5, Zhao Zhihe4,5.   

  1. 1. Dept. of Orthodontics, Hainan Stomatology Hospital, Haikou 570125, China; 2. Dept. of Orthodontics, Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat-sen University, Guangzhou 510055, China; 3. Guangdong Provincial Key Laboratory of Stomatology, Guangzhou 510055, China; 4. Dept. of Orthodontics, West China School of Stomatology, Sichuan University, Chengdu 610041, China; 5. State Key Laboratory of Oral Diseases, Sichuan University, Chengdu 610041, China
  • Received:2011-03-11 Revised:2011-09-10 Online:2012-10-01 Published:2012-10-01

摘要:

目的对比在张应力和压应力两种不同性质力的刺激下人牙周膜成纤维细胞(HPDLF)增殖活性的变化,并对差异表达基因进行筛选,探讨不同应力作用后HPDLF增殖变化的分子机理。方法使用四点弯曲加力装置对细胞进行0.5 Hz、4 000 μstrain的应力加载,加载时间2 h,采用流式细胞术测定张、压应力刺激对HPDLF增殖活性的影响,应用基因芯片技术检测两种应力刺激下细胞的差异表达基因。结果在两种应力刺激作用下,S期细胞百分比和细胞增殖指数均降低;差异表达的基因最主要位于细胞核,集中在转录因子活性相关基因群,参与最多的生物过程为转录因子调控,其中压应力引起的差异表达基因和生物过程的数量较张应力更多。结论1)周期性张、压应力作用下,HPDLF增殖变缓、细胞周期阻滞。2)细胞增殖变缓和细胞周期阻滞可看成是细胞对外界刺激的适应和自我保护,有利于HPDLF有更多的时间决定如何应答外界应力刺激,其应答的结果首先是转录水平上基因表达的改变,最主要的生物学反应是核转录调控。3)HPDLF对周期性压应力更敏感。

关键词: 人牙周膜成纤维细胞, 张应力, 压应力, 增殖, 基因芯片, 差异表达基因

Abstract:

Objective The study was aimed to provide insights into cell proliferation ability alteration of human
periodontal ligament fibroblasts(HPDLF) under two different types of stimulation, cyclic tensile and compressive stress, and screen the differentially expressed genes to have a comprehensive understanding of the molecular mechanism of changes in proliferation. Methods Primary HPDLF were subjected to cyclic tensile and compressive stress(0.5 Hz, 4 000 μstrain) for 2 h through a four-point bending strain system. After that, cell proliferation ability was examined by flow cytometry. Then the gene expression profile was investigated by microarray analysis. Results Mechanical stress slowed a down proliferation of cells. S phase cells and proliferation index(PI) decreased. Differentially expressed genes were mainly located at nucleus and focus in transcription factor activity related genes, and most of them were involved in the regulation of transcription factors. Among them, more were in group of compressive stress. Conclusion 1)Under mechanical loading, HPDLF proliferation were slowed and cell cycles were arrested. 2)Cell cycle arrest and the slowdown proliferation, which can be viewed as an adaptation and protection mechanism of cells to have more time to decide how to respond to mechanical stimuli, and the differential gene expression reflect the behavior of cells. The result of the response is gene transcription changes at first, and the most important biological response is the nuclear transcription. 3)HPDLF are more sensitive to cyclic compressive stress.

Key words: human periodontal ligament fibroblasts;tensile stress; compressive stress; proliferation, microarray; differentially expressed genes