骨髓基质细胞及血管内皮祖细胞共移植对局灶性脑缺血大鼠系列生长因子表达的影响

Expression of growth factors in rats with focal cerebral ischemia after transplantation of bone marrow stromal cells and endothelial progenitor cells

目的观察经静脉移植骨髓基质细胞（BMSCs）及血管内皮祖细胞（EPCs）后血管内皮生长因子（VEGF）、基质细胞衍生因子-1（SDF-1）、碱性成纤维生长因子（bFGF）、胰岛素样生长因子-1（IGF-1）、转化生长因子β（TGF—β）、血小板源性生长因子（PDGF）、脑源性生长因子（BDNF）、胶质源性生长因子（GDNF）和神经生长因子（NGF）这9种细胞因子的表达。方法80只健康成年Wistar大鼠按照随机数字表法分为模型对照组、BMSCs移植组、EPCs移植组及联合移植组，每组20只。线栓法建立大鼠大脑中动脉闭塞模型。造模24h后，分别取浓度为3×10^6个／mL的BMSCs、EPCs悬液及两者混合液1mL经鼠尾静脉移植入BMSCs移植组、EPCs移植组及联合移植组；模型对照组鼠尾静脉注射1mL生理盐水。移植后7d，实时荧光PCR、Westem boltting检测各组大鼠脑内生长因子的表达。结果联合移植组中bFGF、VEGF、BDNFmRNA表达水平最高，与其他3组比较差异均有统计学意义（P〈0．05）。BMSCs移植组中NGF、GDNF、TGF—BmRNA表达水平最高，与其他3组比较差异均有统计学意义（P〈0．05），联合移植组次之。EPCs组中PDGF．BB、IGF-1、SDF-1 mRNA表达水平最高，与其他3组比较差异均有统计学意义（P〈0．05），联合移植组次之。结论BMSCs及EPCs联合静脉移植可上调脑内生长因子表达水平，为提高细胞移植效率提供了新的尝试途径。

Objective To observe the expression of growth factors （vascular endothelial growth factor [VEGF], stromal cell-derived factor-1 [SDF-1], basic fibroblast growth factor [bFGF], insulin-like growth factor [IGF-1], transforming growth factor-β [TGF-β], platelet-derived growth factor [PDGF], brain derived neurotrophic factor [BDNF], glial cell line-derived neurotrophic factor [GDNF] and nerve growth factor [NGF]） in rat ischemic brain tissues after intravenous implantation of bone marrow stromal cells （BMSCs） and/or endothelial progenitor cells （EPCs）. Methods Healthy adult Wistar rats were randomly divided into 4 groups： vehicle group, BMSCs transplantation group, EPCs transplantation group and BMSCs combined with EPCs transplantation group （n=20）. The rats were subjected to middle cerebral artery occlusion （MCAO）, and 24 h after that, they were intravenously transplanted with either 3×10^6 BMSCs, EPCs, BMSCs/EPCs or 1 mL physiological saline. Seven d after transplantation, real time-PCR and Western blotting were employed to detect the expressions ofVEGF, SDF-1, bFGF, IGF-1, TGF-β, PDGF-BB, BDNF, GDNF and NGF. Results The mRNA expressions of bFGF, VEGF and BNDF in the BMSCs/EPCs transplantation group were significantly higher as compared with those in the other groups （P〈0.05）. BMSCs transplantation group enjoyed the highest mRNA levels of NGF, GDNF and TGF-β among all the groups, significantly higher as compared with those in the other groups （P〈 0.05）, followed by BMSCs/EPCs transplantation group. EPCs transplantation group enjoyed the highest mRNA levels of PDGF, IGF-1 and SDF-1, significantly higher as compared with those in the other groups （P〈 0.05）, followed by BMSCs/EPCs transplantation group. Conclusion BMSCs combined with EPCs implantation can promote the fi.mctional rehabilitation in rats after focal cerebral ischemia, which provides new way for improving the transplantation success rate.