布比他丁促进大鼠骨髓间充质干细胞CXC型趋化因子受体4膜向分布及迁移能力的研究

Blebbistatin Promotes the Membrane Distribution of CXC Chemokine Receptor Type 4 and Migration of Rat Bone Marrow Mesenchymal Stem Cells

目的:研究药物布比他丁(Blebbistatin)能否调节大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)表面CXC型趋化因子受体4(CXC chemokine receptor type 4,CXCR4)向胞膜分布,从而促进大鼠BMSCs的迁移。方法:分离培养SD大鼠BMSCs,Blebbistatin处理细胞15、30、45、60min,设置对照组;流式细胞术测定细胞表面CXCR4表达;Transwell迁移实验检测细胞迁移能力;测定成骨诱导前期培养基碱性磷酸酶(alkaline phosphatase,ALP)活性,待诱导完成进行茜素红染色。结果:处理组细胞表面CXCR4表达率明显升高(P<0.01),且呈时间依赖性;15、30和60min组细胞迁移效率升高(P<0.01)。成骨诱导期间各组间ALP活性比较差异无统计学意义,诱导至21d时各组均形成大量钙化结节。结论:Blebbistatin能促进BMSCs表面CXCR4向胞膜分布,并能提高BMSCs的迁移效率。

Objective:To investigate whether Blebbistatin could regulate the distribution of CXC chemokine receptor 4(CXCR4)in rat bone marrow mesenchymal stem cells and promote the migration of bone marrow mesenchymal stem cells(BMSCs).Methods:BMSCs were isolated from SD rats using the whole bone marrow adherence method and the cells were treated with Blebbistatin for 15,30,45 and 60 min.The expression of surface CXCR4 was detected by flow cytometry.Cell migration ability was tested by Transwell migration assay.Osteogenesis was induced then ALP activity was detected in the early stage of induction,and was confirmed by alizarin red staining when induction was completed.Results:The surface expression of CXCR4 of Blebbistatin-treated cells was significantly increased in a time-dependent manner(P<0.01).Cell migration efficiency of Blebbistatin-treated cells increased in 15,30 and 60 min groups(P<0.01).During osteogenic induction,there was no significant difference in the ALP activity among the groups.A large number of calcified nodules were found in all the groups on the 21th day.Conclusion:Blebbistatin promoted the surface expression of CXCR4 in BMSCs and promoted the migration of BMSCs.