华西口腔医学杂志

• 基础研究 • 上一篇    下一篇

Indian Hedgehog在牵张力促进成骨细胞增殖中的作用研究

韩磊1 张晓玲2 唐国华3   

  1. 1.南京大学医学院附属口腔医院口腔正畸科, 南京210025;2.中国科学院上海生命科学研究院/上海交通大学医学院健康科学研究所骨科细胞与分子生物学实验室;3.上海交通大学医学院附属第九人民医院口腔正畸科, 上海市口腔医学重点实验室, 上海200011
  • 收稿日期:2012-06-25 修回日期:2012-06-25 出版日期:2012-06-01 发布日期:2012-06-01
  • 通讯作者: 唐国华,Tel:021-23271699-5206
  • 作者简介:韩磊(1983—),男,山西人,住院医师,硕士
  • 基金资助:

    上海市科学技术委员会基金资助项目(08DZ2271100);上海市重点学科建设基金资助项目(S30206)

Indian Hedgehog signaling is involved in the stretch induced proliferation of osteoblast

Han Lei1, Zhang Xiaoling2, Tang Guohua3.   

  1. 1. Dept. of Orthodontics, Institute and Hospital of Stomatology, Nanjing University Medical School, Nanjing 210025, China; 2. Orthopaedic Cellular and Molecular Biology Laboratory, Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences and Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China; 3. Dept. of Orthodontics, Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai Key Laboratory of Stomatology, Shanghai 200011, China
  • Received:2012-06-25 Revised:2012-06-25 Online:2012-06-01 Published:2012-06-01
  • Contact: Tang Guohua,Tel:021-23271699-5206
  • About author:Han Lei1(1983—),男,山西人,住院医师,硕士

摘要:

目的观察Hedgehog(Hh)信号在牵张力调控成骨细胞增殖中的作用。方法体外分离和培养新生大鼠颅顶骨成骨细胞,应用Flexcell 4000TM加力系统施加3%和6%的2种牵张应变。用N端Hedgehog重组蛋白(N-Shh)、Hedgehog通道抑制剂环巴胺(cy)和机械通道抑制剂三氯化钆(GdCl3)对成骨细胞进行干预。分别采用细胞计数、甲基噻唑基四唑(MTT)比色法和流式细胞仪检测成骨细胞的增殖。采用SAS 8.0软件包对结果进行统计分析。结果在成骨细胞体外培养过程中,牵张力促进了成骨细胞的增殖,以6%牵张应变更显著;牵张力仍能促进经cy处理后的成骨细胞的增殖,但这种促进作用能被GdCl3抑制。结论牵张力促进成骨细胞增殖的作用有一部分是通过上调Ihh的表达实现的。

关键词: 成骨细胞, Hedgehog, 牵张, 增殖

Abstract:

Objective To observe the role of the Hedgehog(Hh) genes in the proliferation of osteoblasts upon mechanical tensile strains. Methods Primary osteoblasts harvested from newborn rat calvarial bone were subjected to 3% and 6% elongation of tensile stretches using Flexcell 4000TM strain unit. The cultures were also treated with either recombinant N-terminals Sonic Hedgehog(N-Shh) or cyclopamine(cy), a Hh inhibitor or gadolinium(GdCl3 ) , an inhibitor of stretch-activated channels. The proliferation of osteoblasts was quantified by cell counting, methyl thiazolyl tetrazolium(MTT) and cell cycle detection via flow cytometry. Statistical analysis was performed using SAS 8.0 software package. Results The tensile strain, especially under 6% elongation, promoted osteoblast proliferation. Stretching force could also promote the proliferation even when the cells were treated with cy, but this effect was suppressed by GdCl3. Conclusion The induced proliferation of osteoblasts by mechanical stretched is mediated at least in part by Indian Hedgehog(Ihh) signaling.

Key words: osteoblast, Hedgehog, stretch, proliferation