华西口腔医学杂志

• 基础研究 • 上一篇    下一篇

张应力诱导大鼠骨髓间充质干细胞骨向分化及其差异基因表达分析

高莺1 李继华1 韩立赤2 马永清1 胡静1 曲丹1 徐誉纯1   

  1. 1.口腔疾病研究国家重点实验室, 四川大学, 四川成都610041;2.大连大学医学院口腔系, 辽宁大连116622
  • 收稿日期:2009-04-25 修回日期:2009-04-25 出版日期:2009-04-20 发布日期:2009-04-20
  • 通讯作者: 李继华,Tel:028-85502334
  • 作者简介:高莺(1975-),女,山西人,博士
  • 基金资助:

    国家自然科学基金资助项目(30400506)

Osteoblastic differentiation and gene expression profile change in rat bone marrow mesenchymal stem cells after a single period of mechanical strain

GAO Ying1, LI Ji-hua1, HAN Li-chi2, MA Yong-qing1, HU Jing1   

  1. 1. State Key Laboratory of Oral Diseases, Sichuan University, Chengdu 610041, China; 2. Dept. of Stomatology, Dalian Medical University, Dalian 116622, China
  • Received:2009-04-25 Revised:2009-04-25 Online:2009-04-20 Published:2009-04-20
  • Contact: LI Ji-hua,Tel:028-85502334

摘要:

目的探讨SD大鼠骨髓间充质干细胞(MSCs)在张应力作用下成骨向分化及其相关基因的差异表达。方法密度梯度离心法体外分离培养大鼠骨髓MSCs;应用四点弯曲加力系统对细胞施加2 000 με的机械牵张力,骨钙素免疫组化染色、碱性磷酸酶(ALP)活力测定MSCs骨向分化,cDNA芯片技术对加力前后MSCs进行mRNA检测,通过芯片杂交、生物信息处理,分析二者间基因差异表达。结果MSCs经应力加载后,其细胞生长曲线与对照组相比差别不大,但骨钙素表达水平和ALP活性显著增高,27 K Rat Genome Array芯片发现,加载前后MSCs表达差异基因共有416条(2.8%),其中表达增强247条(61条显著增强),表达降低169条(74条显著降低)。结论张应力可诱导MSCs骨向分化,而差异表达的基因可能在这一过程中起重要作用。

关键词: 骨髓间充质干细胞, 张应力, 基因芯片

Abstract:

Objective To evaluate the osteoblastic differentiation and compare the difference in the gene expression of rat bone marrow mesenchymal stem cells(MSCs) affected by a single period of mechanical strain. Methods Bone marrow MSCs were harvested from the femurs and tibiae of SD rats and cultured in vitro. A four-point bending apparatus were used to perform a single 40-minute period of 2 000 με mechanical strain on these MSCs  The proliferation of the MSCs was tested by MTT on scheduled date, and the osteoblastic differentiation of the MSCs was measured by testing the expression of osteocalcin and alkaline phosphate(ALP) activity of these cells. In addition, we have investigated the possible mechanisms underlying the action of the single 40 -minute period of 2 000 με mechanical strain on these MSCs, after profile blotted and handled by bioinformation, the gene expressions of these two periods of MSCs were examined. Results The MSCs have grown well in vitro. Our experiment showed that mechanical environment did not weaken the proliferation of the MSCs. However, the ALP activity and the expression of osteocalcin were significantly up-regulated by the 2 000 με mechanical strain. Using the 27 K Rat Genome Array, 416 different expressions were found. The rate of different genes was 2.8%, of which the expressions of 247 genes increased(61 genes remarkably increased) and 169 genes decreased(74 genes remarkably decreased) in these two periods of MSCs. Conclusion Mechanical strain induced the osteoblastic differentiation of the MSCs, which may be attributed to the different gene levels.

Key words: mesenchymal stem cells, mechanical strain, gene chip