成纤维细胞生长因子2(FGF-2)诱导大鼠骨髓间充质干细胞骨架重塑

Cytoskeleton remodeling of rat bone marrow mesenchymal stem cells induced by fibroblast growth factor 2

目的探讨成纤维细胞生长因子2(FGF-2)对大鼠骨髓间充质干细胞(BMSC)形态和骨架的影响。方法采用大鼠动脉内皮细胞(RAEC)和BMSC的TranswellTM隔离共培养体系,通过扫描电镜和罗丹明标记的鬼笔环肽染色观察BMSC的形态和骨架变化,ELISA检测细胞上清中的FGF-2含量;实时定量PCR检测单独和共培养后两种细胞的FGF-2 mRNA水平,共培养体系中添加FGF-2受体的抑制剂NVP-BGJ398,阻断FGF-2受体观察其对BMSC骨架的影响;单独BMSC培养基中添加重组FGF-2,验证外源性FGF-2的作用效果。结果与RAEC共培养后,BMSC逐渐拉伸、收缩,产生大量丝状伪足;在共培养体系中FGF-2含量增加且主要由RAEC分泌;FGF-2受体的抑制剂NVP-BGJ398可显著阻断BMSC骨架重塑,细胞多为短梭形,呈旋涡状排列。外源性FGF-2则促进BMSC胞体收缩、边缘拉伸,产生丝状伪足交错分布。结论RAEC分泌FGF-2诱导BMSC骨架重塑。

Objective To investigate the effects of fibroblast growth factor 2(FGF-2)on the cytoskeleton and morphology of rat bone marrow mesenchymal stem cells(BMSCs).Methods Morphological and cytoskeleton changes of BMSCs were observed by scanning electron microscopy and rhodamine-phalloidin staining in TranswellT M co-culture system of rat vascular endothelial cells(RAECs)and BMSCs.The content of FGF-2 in cell supernatants were detected by ELISA,and the mRNA expression of FGF-2 in both conventional and co-cultured cells were evaluated by real-time quantitative PCR.NVP-BGJ398,an inhibitor of FGF-2 receptor was added into the co-culture system to block FGF-2 signal and its effect on BMSCs skeleton was observed.Recombinant FGF-2 was supplemented into the conventional medium of BMSCs to further verify the effect of exogenous FGF-2.Results After co-cultured with RAECs,BMSCs gradually stretched,contracted and formed a large number of filopodia.The content of FGF-2 increased in the co-culture system and was mainly secreted by RAECs.Cytoskeleton remodeling of BMSCs was significantly blocked by the inhibitor of FGF-2 receptor and the cells were mostly short spindle-shaped and arranged in a spiral pattern.Exogenous FGF-2 promoted the contraction and edge stretching of BMSCs,forming filopodia with staggered distribution.Conclusion FGF-2 secreted by RAECs induces cytoskeletal remodeling of BMSCs.