小鼠外周血和胚胎着床位点中骨髓来源内皮祖细胞在不同妊娠时期的分布和变化规律

Distribution and pattern of endothelial progenitor cell in mouse’s peripheral blood and implantation site during different period of gestation

目的探究小鼠外周血和胚胎着床位点骨髓来源内皮祖细胞(endothelialprogenitor cell,EPC)在不同妊娠时期的分布和变化规律。方法利用5-氟尿嘧啶(5-FU)制备非性腺毒性骨髓抑制小鼠模型并测定其外周血白细胞(WBC)、血小板(PLT)变化以确定小鼠模型建立成功与否,用内皮细胞特异性酪氨酸激酶-2启动子调控下表达绿色荧光蛋白基因(Tie2-GFP)小鼠作为供体并用荧光显微镜观察外周血中有无GFP表达以确定骨髓移植成功与否,将其骨髓移植给骨髓抑制模型小鼠,ELISA方法检测不同妊娠时期外周血血管内皮生长因子(VEGF)含量变化,流式细胞仪测定不同妊娠时期外周血和子宫胚胎着床部位的含量,Pearson相关性分析比较外周血VEGF与骨髓来源EPCs的关系,免疫荧光组织化学染色来观察妊娠不同时间时骨髓来源EPCs在子宫脱膜层血管上的分布。结果小鼠外周血VEGF和EPCs含量在妊娠前期时达到顶点然后逐渐下降,且小鼠外周血VEGF含量与EPCs含量正相关;小鼠胚胎着床位点EPCs含量随着妊娠过程先升高再下降,在妊娠中期达到顶点;妊娠前,在胚胎着床位点EPCs分布较为分散,在妊娠过程中,妊娠前期组外周血VEGF含量均高于未受孕组、妊娠中期组、妊娠晚期组和产后组(P<0.05);妊娠前期组外周血骨髓来源EPCs含量均高于未受孕组、妊娠中期组、妊娠晚期组和产后组(P<0.05);妊娠期小鼠外周血VEGF含量和骨髓来源EPCs含量呈正相关(r=0.421,P=0.002);妊娠前期组、妊娠中期组小鼠胚胎着床位点骨髓来源EPCs含量均高于未受孕组(P<0.05);未受孕组EPCs分布较为分散,而妊娠前期组、妊娠中期组、妊娠晚期组和产后组EPCs分布开始集中且主要分布于血管上皮细胞周围。结论在外周血VEGF含量与EPCs含量正相关,在外周血和胚胎着床位点EPCs含量的变化与小鼠妊娠期的发展有着紧密的关系,EPCs在胚胎着床位点的分布在小鼠妊娠的不同时期也有所不同,EPCs有潜力作为未来监测妊娠期生理状况的指标。

Objective To research on the distribution and pattern of endothelial progenitor cell(EPC)in mouse’s peripheral blood and implantation site during different period of gestation.Methods 5-Fluorouracil(5-Fu)was used to prepare bone marrow suppression mouse,and WBC and PLT change were detected to prove the bone marrow suppression model.tTransplanttyrosine kinases-2 modulating expressionof greenfluorescent protein mouse’s bone marrow to the bone marrow suppression mouse,and fluorescence microscope was used to detect expression of EPC in its peripheral blood to prove the transplantation of bone marrow.ELISA was usd to detect the content of vascular endothelial growth factor(VEGF)in mouse’s peripheral blood.Measure the content of bone marrow derived EPCs in mouse’s peripheral blood and implantation site during different period of gestation by flow cytometer.Pearson correlation analysis was used to compare VEGF and EPCs in peripheral blood.Detect the distribution of EPCs in implantation site with immunofluorescence technique.Results The content of VEGF and EPCs in mouse’s peripheral blood reached the peak at first trimester and then decreased,and the content of VEGF had positive correlation with content of EPCs.The content of EPCs at mouse’s implantation siteincreased and then decline,reached it’s peak at second trimester.Before gestation,EPCs was distributed separately at the implantation site,and during the gestation EPCs’distribution was gradually concentrated around the vascular epithelial cells.Conclusion VEGF has positive correlation with EPCs in mouse’s peripheral blood,EPCs’content change is closely related to the progression of mouse’s gestation,and the distribution of EPCs in implantation site is different during different period of gestation.EPCs are potential to be the indicator of physiological condition during gestation in the future.