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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 rat bone marrow mesenchymal stem cells (BMSCs). Methods BMSCs were isolated by Percoll density gradient centrifugation, and the surface antigen expression and differentiation were identified. Dunn chamber apparatus was used to study the directional migration of BMSCs. The effect of PI3K inhibitor LY294002 on HGF-induced migration of BMSCs was also investigated. Results BMSCs isolated and cultured were positive for CD29, CD90 and CD106, negative for CD34 and CD45. BMSCs could differentiate into osteoblasts and adipocytes. Dunn chamber migration experiments showed that the migration rate of BMSCs did not change with the addition of HGF in the outer tank, the migration efficiency increased with the increase of HGF concentration, and the cell migration efficiency was significantly increased by 50ng / ml and 100ng / ml HGF; 50ng / ml HGF, there was no change in migration rate and migration efficiency. After pretreatment with 30μmol / LLY294002 for 1h, the directional migration of BMSCs induced by HGF was inhibited. Conclusion HGF can chemotaxis the directional migration of BMSCs, and the PI3K signaling pathway is involved in this process.