华西口腔医学杂志

• 基础研究 • 上一篇    下一篇

骨髓间充质干细胞对机械张应力刺激的反应及转化生长因子-β和胰岛素样生长因子-Ⅱ的基因表达

韩立赤1 戚孟春2 孙红2 胡静3 邹淑娟4 李继华3   

  1. 1.大连大学医学院口腔系, 辽宁大连116622; 2.华北煤炭医学院口腔系, 河北唐山063000;3.四川大学华西口腔医学院口腔颌面外科学教研室; 4.正畸学教研室, 四川成都610041
  • 收稿日期:2009-08-25 修回日期:2009-08-25 出版日期:2009-08-20 发布日期:2009-08-20
  • 通讯作者: 邹淑娟,Tel:028-85501425
  • 作者简介: 韩立赤(1970—),女,辽宁人,副主任医师,博士
  • 基金资助:

    国家自然科学基金资助项目(30772454);四川省科技厅攻关课题资助项目(2006Z09-013)

Response of bone marrow mesenchymal stem cells to mechanical stretch and gene expression of transforming growth factor-β and insulin-like growth factor-Ⅱ under mechanical strain

HAN Li-chi1, QI Mengchun2, SUN Hong2, HU Jing3, ZOU Shu-juan4, LI Ji-hua3   

  1. 1. Dept. of Stomatology,Medical College of Dalian University, Dalian 116622, China; 2. Dept. of Stomatology, North China Coal Medical College, Tangshan 063000, China; 3. Dept. of Oral and Maxillofacial Surgery, West China College of Stomatology, Sichuan University, Chengdu 610041, China; 4. Dept. of Orthodontics, West China College of Stomatology, Sichuan University, Chengdu 610041, China
  • Received:2009-08-25 Revised:2009-08-25 Online:2009-08-20 Published:2009-08-20
  • Contact: ZOU Shu-juan,Tel:028-85501425

摘要:

目的探讨大鼠骨髓间充质干细胞(MSCs)对机械张应力刺激的反应及力学刺激下转化生长因子-β(TGF-β)和胰岛素样生长因子-Ⅱ(IGF-Ⅱ)基因表达的规律。方法分离培养大鼠骨髓MSCs,应用四点弯曲加力系统对细胞施加单一周期的机械张应力刺激(2 000 με,40 min)。检测MSCs细胞增殖及碱性磷酸酶(ALP)活性,并采用实时荧光定量RT-PCR检测TGF-β和IGF-Ⅱ的基因表达。结果机械张应力刺激下,MSCs的增殖活力、ALP活性以及TGF-β和IGF-Ⅱ的基因表达均显著增高。TGF-β和IGF-II的mRNA水平在加力后瞬时达最高水平;与对照细胞比较,分别增加了51.44和8.92倍。除加力后6 h有少许增高外,TGF-β和IGF-Ⅱ的表达随时间逐步下降,并于加力后12 h恢复到对照组水平。结论机械张应力刺激可促进MSCs增殖,提高其ALP活性,使TGF-β和IGF-Ⅱ基因表达呈时间依赖性上调,并最终诱导MSCs向成骨细胞分化。机械力学刺激是MSCs骨向分化的关键驱动因子,对牵张成骨骨痂形成具有重要的作用。

关键词: 间充质干细胞, 机械牵张, 转化生长因子-β, 胰岛素样生长因子-Ⅱ

Abstract:

Objective To study the response of rat bone marrow mesenchymal stem cells(MSCs) to a single period of mechanical strain and expression patterns of transforming growth factor-β(TGF-β) and insulin-like growth factor- Ⅱ(IGF-Ⅱ) after mechanical stretch. Methods Bone marrow MSCs were isolated from SD rats and cultured in vitro. A four-point bending apparatus were used to perform a single period of mechanical strain(2 000 με, 40 min) on MSCs. Cellular proliferation and alkaline phosphatase(ALP) activity of MSCs were examined and gene expression patterns of TGF-β and IGF-Ⅱ were detected by SYBR green quantitative real-time RT-PCR. Results Cell proliferation, ALP activity and expression of TGF-β and IGF-Ⅱ were all significantly up-regulated in stretched MSCs when compared with their controls. The mRNA levels of TGF-β and IGF- Ⅱ got top increase immediately after mechanical loading and increased about 51.44 and 8.92 folds, respectively, when compared with control cells. Expression of TGF-β and IGF-Ⅱ decreased with time and returned to control level at 12 h after mechanical stimulus, despite of a small increase at 6 h. Conclusion The mechanical stretch can promote MSCs proliferation, up-regulate its ALP activity and induce a time-dependent expression increase of TGF-β and IGF-Ⅱ which in turn result in osteogenic differentiation of MSCs. Mechanical stimulus is a key stimulator for osteogenic differentiation of MSCs and vital for bone formation in distraction osteogenesis.

Key words: mesenchymalstem cells, mechanicalstretch, transforming growth factor -β, insulin -like growth factor-Ⅱ