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

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

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、CXCR4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CXCR4型受体mRNA、蛋白水平及AKT磷酸化的影响。结果与结论:0.2mg/L的基质细胞衍生因子1α孵育细胞10h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6h;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CXCR4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CXCR4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。

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-la （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-la 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-la 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-la preconditioning. The chemokine receptor CXCR 4 expression and Akt phosphorylation were increased after SDF-la preconditioning. SDF-la preconditioning can enhance the migration of BMSCs through increasing the expression of chemokine receptor CXCR 4 Akt signaling pathway is correlated with this migration.