腺病毒介导的人基质细胞衍生因子-1基因转移促进间充质干细胞的迁移

Adenovirus-mediated stromal cell-derived factor-1 alpha gene transfer promotes mesenchymal stem cell migration

目的初步探索在活体状态下基质细胞衍生因子-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。方法以MSC为实验材料,将纯化的hSDF-1α重组腺病毒100!l加入单个散在分布的MSC培养基中,分别于转染后1,2,3d观察MSC迁移的变化。随后于hSDF-1α重组腺病毒转染MSC后12h,以50"nmol/L渥曼青霉素,10#mol/LLY294002,50$mol/LPD98059,10%mol/LU73122,0.1g/LAMD3100以及50nmol/L维拉帕米等分别处理MSC,观察转染SDF-1的MSC迁移的信号机制。结果转染SDF-1重组腺病毒的MSC呈聚集的集落样生长,渥曼青霉素、LY294002、PD98059、U70312、AMD3100、维拉帕米等处理后,MSC聚集的集落样生长趋势明显降低,尤以U73122、AMD3100对其阻断效应最为显著。结论SDF-1促进了MSC的迁移,其作用与丝裂原活化蛋白激酶(MAPK)、磷脂酰肌醇特异性磷脂酶C(PI-PLC)和蛋白激酶C(PKC)等信号分子有关。

Objective To explore the role ofstromal-derived factor-1 （SDF-1） in the migration ofmesenchyrnal stem cells （MSCs） and the underlying signal transduction mechanism. Methods Rat bone marrow-derived MSCs were infected with 100 ml recombinant adenovirus containing human SDF-la gene （Ad-hSDF-lot,）, and the cell migration changes were observed at 1, 2, and 3 days after the infection. Twelve hours after Ad-hSDF-lot transfection, the MSCs in separate cultures were treated with wortmannin （50 nmol/L）, LY294002 （10 mmol/L）, PD98059 （50 mmol/L）, U73122 （10 mmol/L）, AMD3100 （0.1 g/L）, or verapamil （50 nmol/L）, respectively, and the signal transduction pathways involved in MSC migration were analyzed. Results The MSCs grew in colonies after transfection with Ad-hSDF-1α, but this growth pattern was substantially attenuated after treatmentwithwortmannin, LY294002, PD98059, U73122, AMD3100andverapamil, among whichU73122and AMD3 100 treatments resulted in the most conspicuous inhibitory effects. Conclusion The effect of SDF-1 in promoting MSC migration is related to mitogen-activated protein kinase, phosphatidylinositol phospholipase C, and protein kinase signal pathways.