基质细胞衍生因子-1通过促进 c-kit表达提高小鼠心肌干细胞增殖和迁移能力的研究

摘要

目的：探讨基质细胞衍生因子-1（ SDF-1）提高小鼠心肌干细胞（ CSCs ）增殖和迁移能力的机制。方法：从小鼠心肌中分离、培养出CSCs ，并用免疫磁珠法筛选出c-kit （＋） CSCs ，用流式细胞仪检测CSCs表面标志：干细胞因子受体c-kit、干细胞抗原Sca-1。用SDF-1及SDF-1特异性拮抗剂AMD3100干预c-kit（＋） CSCs，qPCR及Western blotting检测c-kit的mRNA和蛋白表达，用CCK-8试剂盒及transwell趋化实验检测细胞增殖和迁移能力。结果：分离、培养出的细胞经免疫磁珠筛选后，流式细胞仪检测c-kit和Sca-1表达均大于80％。 SDF-1干预后， qPCR和Western blotting 结果显示c-kit mRNA 和蛋白表达均增高， CCK-8及transwell趋化实验结果显示SDF-1干预后细胞的增殖及迁移能力明显提高，并且可以被AMD3100拮抗。结论：SDF-1通过促进c-kit表达提高c-kit（＋） CSCs的增殖及迁移能力。%Objective:To investigate the mechanism on improving proliferation and migration abilities of cardiac stem cells ( CSCs) by stromal cell derived factor-1 ( SDF-1) .Methods:CSCs were isolated from adult mice hearts purified by magnetic-activated c-kit cell sorting magnetic beads .The markers of c-kit and Sca-1 were measured by FACS.The cells were cultured with SDF-1 and CXCR4-selective antagonist AMD3100, and c-kit expression was measured by qPCR and Western blotting .After the intervention of SDF-1 and AMD3100, proliferation and migration of c-kit ( +) CSCs were measured by CCK-8 assay and transwell assay .Results: More than 80%cardiosphere-derived cells which were purified by magnetic-activated c-kit cell sorting magnetic beads were positive for c-kit and Sca-1 by FACS.SDF-1 could enhance the expression of c-kit mRNA and protein, and AMD3100 could inhibit this function.And SDF-1 could enhance the proliferation and migration of c-kit ( +) CSCs, and AMD3100 could inhibit these functions .Conclusion:SDF-1 can improve the proliferation and migration abilities of CSCs by SDF-1 .