hSDF-1α/CXCR4介导骨髓源间充质干细胞迁移的信号转导

Signal transduction related to hSDF-1α/CXCR4-mediated mesenchymal stem cell migration

目的:利用Boyden chamber体外迁移体系初步探索基质细胞衍生因子-1(SDF-1)在骨髓源间充质干细胞(MSC)迁移中的作用及其信号转导机制.方法:采用经典的全骨髓贴壁法培养MSC,通过成骨、成脂肪等多向诱导分化以及流式细胞仪分析其表面标记(CD133,CD34,CD90,CD105)等鉴定MSC特征;以P3-MSC为实验材料,利用Boyden chamber体外迁移体系,观察不同浓度的hSDF-1对MSC迁移的影响,以50nmol渥曼青霉素,10mmol LY294002,50mmol PD98059,10mmol U73122,0.1g/L AMD3100等不同处理P3-MSC,观察最适宜浓度的hSDF-1对MSC迁移影响的信号转导机制.结果:培养的MSC呈现出CD90,CD105强阳性,具有成骨、成脂肪等多向分化能力;MSC体外迁移能力随着hSDF-1α浓度(1,10,100,500μg/L)的递增而逐渐增强,并且hSDF-1α浓度在100μg/L时,MSC迁移到滤膜上的细胞数接近于峰值;渥曼青霉素,LY294002,PD98059,U70312,AMD3100,维拉帕米对MSC迁移均有影响,其中U73122,AMD3100对MSC迁移阻断的效应最显著.结论:SDF-1/CXCR4所介导的MSC迁移与丝裂原活化蛋白激酶(MAPK),磷脂酰肌醇特异性磷脂酶C和蛋白激酶C(PKC)等信号途径有关,且PKC途径可能处于中心环节.

AIM： To explore the role of stromal cell-derived factor-1 （SDF-1） in the migration of mesenchymal stem cell （MSC） and its signal transduction mechanism. METHODS： MSC culture was performed with the classical whole bone marrow adhering method; characteristics of MSC were identified through the induction of multiple-differentiation into osteoblasts or lipoblasts, and surface marker assay （ CD133, CD34, CD105） ; by using Boyden chamber in vitro migration assay system, the effects of SDF-1 concentrations on P3 MSC migration were observed and the signal transduction pathways related to P3-MSC migration were analyzed. RESULTS： Cultured MSC had the potential of menbranedifferentiation and highly expressed CD105 and CD90; the efficiency of MSC migration into the filter membrane increased gradually with the increase of hSDF-lconcentration （1, 10, 100, 500 μg/L）, and MSC number in the filter membrane was approximately maximum when hSDF-1 was 100 μg/L. 50 nmol wortmannin, 10 mmol LY294002, 50 mmol PD98059, 10 mmol U73122 and 0. 1 g/L AMD3100 were all able to decrease the number of MSC in the filter membrane, and what＇s more, U73122 and AMD3100 treatments were most effective in blocking the MSC migration. CONCLUSION： SDF-1/CXCR4-mediated MSC migration is related to mitogen activated protein kinase （ MAPK）, phosphatidylinositol phospholipase C （ PI-PLC ） and protein kinase （PKC） signal pathways, but PKC signal pathway may be the central role in MSC migration.