摘要
目的研究粒细胞—巨噬细胞集落刺激因子对内皮祖细胞功能活性的影响,为探讨血管内皮再生修复机制和心血管疾病的防治提供实验基础。方法首先用密度梯度离心法分离培养大鼠骨髓和脾内皮祖细胞,在倒置相差显微镜下观察内皮祖细胞的生长分化过程。标记FITC-AC133和PE-vWF鉴定内皮祖细胞,在荧光显微镜下观察。然后选取培养7d的内皮祖细胞施加实验因素。分别用MTT比色法、Transwell小室、Matrigel管腔形成的体外模型检测粒细胞—巨噬细胞集落刺激因子对内皮祖细胞增殖、迁移、管腔形成能力的影响。结果与对照组相比,粒细胞—巨噬细胞集落刺激因子作用后,内皮祖细胞的OD490值、迁移的细胞数和形成的管腔数显著增加(P<0.001),且随着作用浓度的增加和作用时间的延长内皮祖细胞的OD490值、迁移的细胞数和形成的管腔数逐渐增加。结论在体外,粒细胞—巨噬细胞集落刺激因子能够增强内皮祖细胞的增殖、迁移和管腔形成能力,并随着作用浓度的增加和作用时间的延长其效应增强。
Aim To investigate the influence of granulocyte-macrophage colony stimulating factor on functional activity of rat endothelial progenitor cells in vitro in order to further provide experimental basis for the mechanism of reendothelialization and prevention and treatment of coronary artery disease, Methods Endothelial progenitor cells were separated from bone marrow and spleen of rat with density gradient centrifugafion method, Then growth and differentiation procession of endothelial progenitor cells were observed with inverted phase contrast microscope. Endothelial progenitor cells were identified via dual fluorescent staining of FITC-AC133 and PE-vWF. After cultured for seven days, endothelial progenitor cells were added stimulating factors. Then the influence of granulocyte-macrophage colony stimulating factor on proliferation, transfer and blood vesels formation of endothelial progenitor cells were detected with MTT colorimetry, Transwell chamber and Matrigel. Finally the experiment results were analysed. Results Compared with control, granuloeyte-macrophage colony stimulating factor significantly increased OD490, the number of migratory cells and tubes （ P 〈 0. 001 ）. Conclusions Granuloeyte-macmphage colony stimulating factor can significantly facilitate proliferation, migration and blood vessels formation of endothelial progenitor cells in vitro.
出处
《中国动脉硬化杂志》
CAS
CSCD
2008年第7期541-544,共4页
Chinese Journal of Arteriosclerosis
