不同迁移体系对SDF-1/CXCR4介导间充质干细胞迁移的影响

目的:利用Boyden chamber体外迁移体系以及活体状态下观察基质细胞衍生因子-1(SDF-1)对间充质干细胞(MSC)迁移的影响。方法:利用Boyden chamber体外迁移体系,先观察不同浓度hSDF-1对MSC迁移的影响,随后利用活体迁移体系,观察MSC转染Ad-hSDF-1后迁移的变化。最后利用上述体系经不同的阻断剂分别处理MSC,观察其对MSC迁移的影响。结果:MSC体外迁移能力随着hSDF-1α浓度递增而逐渐增强,并且Wortmannin、LY294002、PD98059、SB203580、PKC-ζ假底物、U70312、AMD3100、Verapamil对MSC迁移均有影响,其中PKC-ζ假底物、U73122、AMD3100对MSC迁移阻断的效应最显著。结论:SDF-1/CXCR4所介导的MSC迁移与PI3K、MAPK、PI-PLC/PKC等信号途径有关,且PKC途径可能处于中心环节。

Effect of SDF-1 on mesenchymal stem cells migration through different migration assay systems

Objective： To explore the signal transduction mechanism of stromal derived factor-1 （SDF-1） on migration of mesenchymal stem cell （MSC）. Methods： The SDF-1 density-dependent of MSC migration was observed. And the characteristics of MSC migration were observed for 1, 2 and 3 days after Ad- EGFP-hSDF-1α （100μl） was added into1 single-disposition MSC. Subsequently, 12 hours later MSC transfected by Ad-EGFP-hSDF-1α were treated with 50 nmol wortmannin, 10μmol/L LY294002, 50μmol/L PD98059, 10μmol/L, U73122 and 126μmol/L AMD3100 and 50 nmol/L verapamil, respectively. Results：The efficiency of MSC migration increased gradually with the density increasing of hSDF-1, and after MSCs treatment with 50 nmol wortmannin, 10μmol LY294002,50μmol PD98059,30μmol SB203580,50μmol PKC-PI 10μmol U73122 and 126μmol/L AMD3100, respectively, the ability of MSC migration was decreased. Importantly, the ability of MSCs migration obviously decreased when MSCs with U73122, AMD3100 treatment by different migration assay systems. Conclusion：SDF-1/CXCR4-mediated MSCs migration is related with mitogen-activated protein kinase （MAPK）,phosphatidylinositol phospholipase C（PI-PLC） and protein kinase （PKC） signal pathways, PKC signal pathway may be the central role for MSCs migration.