肝细胞生长因子对骨髓间充质干细胞迁移的影响

Effect of Hepatocyte Growth Factor on the Migration of BMSCs

目的研究肝细胞生长因子(HGF)对大鼠骨髓间充质干细胞(BMSCs)迁移的影响。方法采用Percoll密度梯度离心法分离BMSCs,进行表面抗原表达及分化鉴定。使用Dunn chamber装置研究BMSCs的定向迁移。并且研究PI3K抑制剂LY294002对HGF诱导BMSCs迁移的影响。结果分离培养的BMSCs呈CD29、CD90、CD106阳性表达,CD34、CD45阴性表达,BMSCs可诱导分化为成骨细胞及脂肪细胞。Dunn chamber迁移实验显示,外槽加入不同浓度的HGF,BMSCs迁移速率没有变化,迁移效率随着HGF浓度的增加而增加,50ng/ml与100ng/mlHGF均显著提高了细胞迁移效率;内外槽同时加入50ng/mlHGF,迁移速率与迁移效率均没有变化;经30μmol/LLY294002预处理1h后,HGF诱导的BMSCs的定向迁移受到抑制。结论 HGF能够趋化BMSCs的定向迁移,PI3K信号通路参与介导这一过程。

Objective To investigate the effect of hepatocyte growth factor（HGF） on the migration of bone marrow mesenchymal stem cells（BMSCs）.Methods BMSCs were isolated from bone marrow of rats by Percoll gradient centrifugation,characterized by antigen expression and differentiation potential.Dunn chamber was used to observe the migration of BMSCs towards HGF.Moreover,the role of LY294002,a PI3K inhibitor,on the HGF-induced migration of BMSCs was investigated.Results BMSCs were positive for CD29,CD90,CD106 and negative for CD34,CD45.BMSCs could be induced to differentiate into osteoblasts and adipocytes.Dunn chamber analysis showed that HGF at any concentration in the outer well did not change the migration speed of BMSCs,while the migration efficiency of BMSCs increased gradually with the increasing concentration of HGF,both 50 ng /ml and 100 ng /ml HGF raised the migration efficiency significantly.In contrast,neither the migration speed nor the migration efficiency was changed when both inner and outer wells were filled with 50 ng /ml HGF.The HGF-induced directed migration of BMSCs was inhibited by 30 μmol /L LY294002.Conclusion HGF can induce the targeted migration of BMSCs and PI3K signaling pathway participates in this process.