bFGF和PDGF-BB对内皮祖细胞分化的内皮样细胞和平滑肌样细胞增殖和迁移的影响

The Effect of Basic Fibroblast Growth Factor and Platelet-Derived Growth Factor-BB on the Proliferation and Migration of Endothelial Progenitor Cells-Derived Endothelial-Like Cells and Smooth Muscle-Like Cells

目的探讨内皮祖细胞分化的内皮样和平滑肌样细胞在碱性成纤维生长因子和血小板源生长因子BB作用下的增殖和迁移能力。方法将分离、培养及纯化的内皮祖细胞培养5天后进行分组:对照组、碱性成纤维生长因子组和血小板源长因子BB组。对照组使用20%胎牛血清的DMEM培养基;碱性成纤维生长因子组和血小板源生因子BB组在20%胎牛血清的DMEM培养基中分别添加碱性成纤维生长因子(30μg/L)和血小板源生长因子BB(40μg/L)。免疫荧光染色鉴定内皮祖细胞以及检测内皮祖细胞内皮方向分化标记物CD31和vWF,及平滑肌方向分化标记物α-SMA和Calponin。将分化的内皮样细胞和平滑肌样细胞用MTT法和Transwell小室分别检测其增殖活性和迁移能力。结果与对照组比较,内皮祖细胞经碱性成纤维生长因子或血小板源生长因子BB诱导后呈现较强的内皮细胞(CD31,vWF)或平滑肌细胞(α-SMA,Calponin)的荧光染色,被诱导细胞分别称为内皮样细胞和平滑肌样细胞;碱性成纤维生长因子促进内皮样细胞和血小板源生长因子BB促进平滑肌样细胞增殖的作用在一定范围内具有时间(0～48 h)和浓度(碱性成纤维生长因子:0～16μg/L;血小板源生长因子BB:0～18μg/L)依赖效应。结论碱性成纤维生长因子和血小板源生长因子BB分别可以促进内皮祖细胞分化成内皮样细胞和平滑肌样细胞,并且在碱性成纤维生长因子和血小板源生长因子BB作用下具有明显的增殖和迁移能力。

Aim To study the effect of basic fibroblast growth factor（bFGF） and platelet-derived growth factorBB （PDGF-BB） on the proliferation and migration of endothehal progenitor cells （EPC）-derived endothelial-like cells and smooth muscle-like cells. Methods EPC isolated and cultured for 5 days were stained by immunofluorescence and observed with laser confocal microscopy, the cells of which both AC133 and vWF are positive are the differentiating EPC. After 5 days of EPC culture, the medium was replaced by either control medium （ DMEM, supplemented with 20% fetal bovine serum） or control medium containing rat bFGF（30 μg/L） or PDGF-BB （40 μg/L）. The cultures were incubated up to 14^th day. Then, endothelial cell marker （CD31 and vWF） of bFGF-induced cells （FIC） and non-induced cells （NIC） or smooth muscle cell marker （ct-SMA and calponin） of PDGF-BB-induced cells （PIC） and non-induced cells （NIC） were measured by confocal microscopy. The proliferation and migration of the endothelial-like cells and smoothmuscle-like cells-derived EPC were detected by MTT assays and Transwe11. Results Compared with control group, EPCs induced by bFGF （named as endothelial-like cells） or EPC induced by PDGF-BB （named as smooth musclelike cells） showed respectively more significant endothelial mark（CD31 ,vWF） or smooth muscle mark（α-SMA, calponin） ; In the period of 0 -48 h, bFGF and PDGF-BB respectively promoted endothelial-like cells and smooth muscle-like cells proliferation at the corresponding concentration and there were time-dependent and dose-dependent manner. Conclusion bFGF and PDGF-BB may promote EPC to differentiate into endothelial-like cells and smooth muscle-like cells, and these endothelial-like cells and smooth muscle-like cells have the obvious ability of proliferation and migration.