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  

Mechanism investigation on improving proliferation and migration abilities of cardiac stem cells by stromal cell derived factor-1

作　　者：陈中璞[1] 潘啸东[1] 姚玉宇[2] 马根山[2] 

机构地区：[1]东南大学医学院 [2]东南大学附属中大医院心内科

出　　处：《东南大学学报（医学版）》2013年第6期674-679,共6页Journal of Southeast University(Medical Science Edition)

基　　金：国家自然科学基金资助项目(81270204)

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 .

关 键 词：C-KIT 心肌干细胞 基质细胞衍生因子-1 CXCR4 小鼠 

分 类 号：Q7[生物学—分子生物学]