绝经前后妇女外周血内皮祖细胞数量和功能的对比研究

The comparision of the number and activity of endothelial progenitor cells in peripheral blood in premenopausal and postmenopausal women

目的：观察绝经前后妇女血清雌二醇的变化对外周血内皮祖细胞（endothe-lial progenitor cells，EPCs）的数量和功能的影响，评价雌二醇水平和外周血内皮祖细胞数量的相关性。方法：选择45-54岁年龄段绝经前后的妇女23例，其中绝经前组11例，绝经后组12例。用放射免疫法测定血清雌二醇浓度。用密度梯度离心法从外周血获取单个核细胞，将其接种在人纤维连接蛋白包被培养板，培养7天后对贴壁细胞进行化学分析。激光共聚焦显微镜鉴定Dil-acLDL和FITC-UEA-I双染色阳性细胞为正在分化的EPCs。用倒置荧光显微镜进行细胞计数，观察内皮祖细胞的黏附、增殖能力。结果：绝经后组外周血EPCs数量较绝经前组明显减少，两组差异有统计学意义（P〈0．001）；EPCs数量和雌二醇水平呈正相关（r=0．875，P〈0．001）。绝经后妇女EPCs的黏附能力和增殖能力也明显受损。结论：妇女血雌二醇水平与外周血EPCs的数量和功能有一定的内在联系，这可能是其干预心血管病危险因素的机制。

Objective：To investigate whether there was difference between the number and activity of endothelial progenitor cells （EPCs） in both premenopausal and postmenopausal women, and to assess the correlation between serum estradiol levels and the number of EPCs. Methods：Eleven premenopausal women and twelve postmenopausal women both aged within 45 to 54 years old were selected. Serum estradiol levels were tested by radioimmunoassay（RIA） in 23 women. Mononuclear cells were isolated from peripheral blood in 23 women by Histopaque density gradient centrifugation, and then the cells were plated on fibronectin-coated culture dishes. At day 7 of culture, the immunohistochemical characterization of the adherent cells was performed. EPCs were characterized as adherent cells which were double positive for Dil-acLDL-uptake and FITC-UEA-I binding by direct fluorescent staining demonstrated under a laser scanning confocal microscope and evaluated the number of EPCs with an inverted fluorescent microscope. EPCs proliferation activity were assayed with MTT assay. EPCs adhesion assay was also performed by replating cells on fibronectin-coated dishes, then counting the adherent cells to value the adhesive capacity. Results： The number of EPCs in postmenopausal women was significantly decreased as compared with that in premenopausal women （P 〈 0.001 ）, and the number of EPCs was positively correlated with the serum estradiol levels （r = 0.875 ,P 〈 0.001 ）. In addition,EPCs adhesive and proliferative capacity were also impaired （P 〈 0. 001 ）. Conclusion： The serum estradiol levels of females are correlated with the number and the part functions of EPCs,which can modulate the cardiovascular risk.