中国组织工程研究

中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (27): 5046-2051.doi: 10.3969/j.issn.2095-4344.2012.27.020

• 干细胞因子及调控因子 stem cell factors and regulatory factors • 上一篇    下一篇

基质细胞衍生因子1α预处理大鼠骨髓间充质干细胞的迁移

束 波1，刘志江2，范 芳1   

  1. 1遵义医学院生物化学与分子生物学教研室，贵州省遵义市 563003；
    2遵义医学院第一附属医院心内科，贵州省遵义市 563003
  • 收稿日期:2012-03-01 修回日期:2012-04-10 出版日期:2012-07-01 发布日期:2013-11-01
  • 作者简介:束波★，女，1980年生，贵州省遵义市人，汉族，2008年上海交通大学医学院毕业，硕士，讲师，主要从事干细胞移植的基础研究。 shubo1014@126.com
  • 基金资助:

    贵州省科学技术基金资助项目(黔科合J字LKZ[2010]29号)

Effect of stromal cell derived factor-1α preconditioning on migration of rat bone marrow mesenchymal stem cells

Shu Bo1, Liu Zhi-jiang2, Fan Fang1   

  1. 1Department of Biochemistry and Molecular Biology of Zunyi Medical College, Zunyi 563003, Guizhou Province, China;
    2Department of Cardiology, First Affiliated Hospital of Zunyi Medical College, Zunyi 563003, Guizhou Province, China
  • Received:2012-03-01 Revised:2012-04-10 Online:2012-07-01 Published:2013-11-01

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摘要：

背景：骨髓间充质干细胞移植过程中只有少量细胞能定向迁移到损伤组织，因此如何提高细胞定向迁移的数量是干细胞移植的关键因素。
目的：观察基质细胞衍生因子1α预处理对大鼠骨髓间充质干细胞定向迁移的影响。
方法：体外分离培养骨髓间充质干细胞，予以传代。使用Transwell体外迁移体系，观察不同浓度的基质细胞衍生因子1α趋化骨髓间充质干细胞定向迁移，选取最佳浓度值趋化细胞。在基质细胞衍生因子1α预处理、CXCR 4型受体阻断剂AMD3100和Akt通路阻断剂LY294002的干预下观察骨髓间充质干细胞的趋化迁移情况，检测基质细胞衍生因子1α预处理对骨髓间充质干细胞中CXCR 4型受体 mRNA、蛋白水平及AKT磷酸化的影响。
结果与结论：0.2 mg/L的基质细胞衍生因子1α孵育细胞10 h具有最佳趋化细胞迁移效应。骨髓间充质干细胞的定向趋化迁移作用随着基质细胞衍生因子1α预处理细胞浓度的增加而增强，预处理时间为6 h；而AMD3100和LY294002可阻断基质细胞衍生因子1α预处理的促迁移作用；基质细胞衍生因子1α预处理可上调骨髓间充质干细胞CXCR 4型受体的表达并促进Akt蛋白磷酸化。提示基质细胞衍生因子1α预处理可通过增加骨髓间充质干细胞表面的CXCR 4型受体数量，从而增强基质细胞衍生因子1α/CXCR4型受体介导的骨髓间充质干细胞定向迁移，该预处理效应可能与Akt信号途径有关。

关键词: 基质细胞衍生因子1α, 骨髓间充质干细胞, 预处理, 迁移, CXCE 4型受体, Akt信号, 干细胞

Abstract:

BACKGROUND: During transplantation of bone marrow mesenchymal stem cells (BMSCs), only a few BMSCs can migrate into injured tissue. Therefore, how to increase the number of migrated BMSCs is a key to BMSC transplantation.
OBJECTIVE: To investigate the effect of stromal cell derived factor-1α (SDF-1α) preconditioning on the migration of rat BMSCs.
METHODS: BMSCs were in vitro isolated, cultured, and passaged. Using Transwell inserts technique, the effects of SDF-1α at different concentrations on the migration of BMSCs were observed and the optimal concentration of SDF-1α was determined. After preconditioned by SDF-1α, SDF-1α+AMD3100 and SDF-1α+LY294002 respectively, the migration of BMSCs was observed, and the effect of SDF-1α preconditioning on mRNA and protein level of chemokine receptor CXCR 4 in BMSCs and AKT phosphorylation were investigated.
RESULTS AND CONCLUSION: Incubation of 0.2 mg/L SDF-1α for 10 hours could produce the optimal migration of BMSCs. The role of SDF-1α in inducing the migration of BMSCs increased with the increasing concentration of SDF-1α used for 6 hours of preconditioning. AMD3100 and LY294002 could reverse the effect of SDF-1α preconditioning. The chemokine receptor CXCR 4 expression and Akt phosphorylation were increased after SDF-1α preconditioning. SDF-1α preconditioning can enhance the migration of BMSCs through increasing the expression of chemokine receptor CXCR 4. Akt signaling pathway is correlated with this migration.

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