目的观察骨髓内皮祖细胞(EPC)移植对假孕大鼠黄体促血管生长因子(VEGF)表达的影响,探讨骨髓EPC促进假孕大鼠黄体血管新生的作用机制。方法雌性大鼠经137 Cs全身照射后,尾静脉移植雄性大鼠骨髓EPC行造血重建,然后制备假孕模型为移植假孕组;分别于假孕第3、7、11、15天应用real time PCR法检测黄体VEGF、碱性成纤维细胞生长因子(bFGF)mRNA的表达;蛋白免疫印迹法(Western blot)检测黄体VEGF、bFGF蛋白表达;CD31免疫组织化学法检测黄体微血管密度。结果假孕对照组(输注等量培养液)及移植假孕组VEGF、bFGF表达在第7天达到最高;黄体微血管密度在第7、11天均高于第3天组水平(P〈0.01);移植假孕组VEGF、bFGF表达在第7、11天均高于同期假孕对照组(P〈0.05),且黄体微血管密度在第7、11天较同期假孕对照组高(P〈0.05)。结论骨髓EPC移植后黄体VEGF、bFGF mRNA及其蛋白表达水平升高,促进假孕大鼠黄体血管新生。
Objective To observe the effection of bone marrow endothelial progen-itor cells(EPCs) transplantation on corpus luteum angiogenic growth factors expression,and investigate the mechanism of the EPCs to promote the angiogenesis of pseudo- pregnancy rat corpus luteum. Methods Female rats radiated by lSTCs were transplanted EPCs which came from male rat via tail vein. The pseudocyesis model of transplantated female rat was established after hematopoietic reconstitution,which named as pseudopregnancy group. On 3rd, 7th, llth and 15th day,Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) mRNA were tested through the method of RT-PCR,their protein expression were tested through the method of Western blotting, and the the corpus luteum capillary density were tested through CD31 immunohisto chemistry. Results In pseudopregnancy control groups and transplantation pseudopr-egnancy groups, the VEGF. bFGF expression achieved the peak on 7th day. The eorp-us luteum capillary density was higher than that of 3rd day group(P〈0. 01)on the 7th and llth day group. The expression of VEGF and bFGF of transplantation pseudopregnancy group were higher than that of pseudopregnancy control group on 7th, 11th day(P〈0.05), the corpus luteum capillary density of transplantation pseudopregnancy group were higher than that of pseudopregnancy control group on 7th and 11th day(P〈0.05). Conclusion EPCs transplantation can increase the expression of VEGF and bFGF mRNA,and promote the growth of corpus luteum angiogenesis.