目的探讨血小板衍生生长因子BB(PDGFBB)对大鼠骨髓间充质干细胞(BMSCs)定向迁移的影响及其在BMSCs向C6胶质瘤迁移过程中的作用。方法直接贴壁法分离培养传代BMSCs,RT-PCR检测PDGF受体α和β mRNA在BMSCs的表达,用Transwell小室建立体外迁移模型检测PDGFBB对BMSCs迁移的影响及其在BMSCs向C6胶质瘤的迁移过程中的作用,评价p38丝裂原蛋白激酶(p38MAPK)抑制剂SB203580对PDGFBB诱导BMSCs迁移能力改变的影响。结果通过直接贴壁法获得了纯化的BMSCs,RT-PCR证实PDGF受体α和βmRNA在BMSCs呈阳性表达,C6胶质瘤可以诱导BMSCs向胶质瘤细胞定向迁移。用阻断抗体阻断PDGFBB后,向C6胶质瘤发生迁移的BMSCs数量减少;含10ng/mlPDGFBB的无血清培养基可诱导BMSCs迁移;加入SB203580后PDGFBB诱导发生迁移的BMSCs数量减少。结论 PDGFBB是诱导BMSCs向C6胶质瘤发生迁移的细胞因子之一。PDGFBB可诱导BMSCs发生迁移,p38M APK参与了这一过程的信号转导。
Objective To explore the effect of platelet-derived growth factor BB (PDGFBB) on the directional migration of rat bone marrow-derived mesenchymal stem cells(BMSCs) and its role in the migration of BMSCs towards C6 glioma. Methods BMSCs were isolated, cultured and passaged by direct adherent culture method. The expression of PDGF recepter αand β mRNA in BMSCs was evaluated by Reverse Transcription-PCR. Using transwell inserts technique, we established in vitro model to study the impact of PDGFBB on the migration of BMSCs and its effect on the migration of BMSCs towards C6 glioma and to explore the regulatory role of SB203580, a p38 mitogen-activated protein kinase inhibitor, in the PDGFBB-induced migrating capacity change of BMSCs. Results BMSCs were successfuly cultured and purified by direct adherent culture method. The results of RT-PCR confirmed that BMSCs expressed PDGF recepter αand β mRNA. C6 glioma cells could induce BMSCs to migrate towards themselves, blocking of PDGFBB by a neuturalizing antibody inhibited the migration of BMSCs towards C6 glioma. Serum-free medium containing 10 ng/ml PDGFBB could induce the migration of BMSCs, SB203580 decreased the number of BMSCs migrating towards PDGFBB. Conclusion PDGFBB contributes the migration of BMSCs induced by C6 glioma and induces the migration of BMSCs, p38MAPK participates in the intracellular signal transduction of this migration.