雷帕霉素及基质细胞衍生因子1α对内皮生长晕细胞生物学功能的影响

The Effects of Rapamycin and Stromal Cell Derived Factor-1α on the Biological Function of Endothelial Outgrowth Cell

目的观察基质细胞衍生因子1α对内皮生长晕细胞生物学功能的作用,以及探讨雷帕霉素对这种作用的影响。方法分离脐血中单个核细胞,采用贴壁法培养内皮生长晕细胞。免疫细胞化学染色鉴定第二代细胞的内皮前体细胞特性。取第3代细胞分为四组,分别用10μg/L基质细胞衍生因子1α、10μg/L基质细胞衍生因子1α+1μg/L雷帕霉素、1μg/L雷帕霉素、普通培养液(空白对照)孵育内皮生长晕细胞24 h。采用CCK-8法检测内皮生长晕细胞的增殖能力,划痕试验检测内皮生长晕细胞的迁移能力。结果采用贴壁法培养的细胞具有多种内皮细胞特性。与10μg/L基质细胞衍生因子1α共同孵育的内皮生长晕细胞其增殖能力和迁移能力均得到活化(P<0.01),但1μg/L雷帕霉素抑制了这种活化作用(P<0.01)。并且1μg/L雷帕霉素能够明显抑制内皮生长晕细胞的生物学功能(P<0.05)。结论基质细胞衍生因子1α能够活化内皮生长晕细胞的增殖能力及迁移能力,雷帕霉素不但抑制内皮生长晕细胞本身的迁移能力,而且能够抑制内皮生长晕细胞的增殖能力并抵消基质细胞衍生因子1α对内皮生长晕细胞生物学功能的活化作用。

Aim To observe the role that stromal cell derived factor-lα （SDF-lα） plays in the biological functions of endothelial outgrowth cell （EOC）, and the effect that rapamycin exerts on such role. Methods The mononuclear cells were harvested from umbilical cord blood by Ficoll density gradient centrifugation, induced into EOC and then expanded in vitro. The endothelial progenitor characteristics of second generation EOC were identified by immunostalning staining. The third generation EOC were devided into 4 groups, which were treated with 10 μ/L SDF-lα, 10μg/L SDF-lα ＋ 1 μg/L rapamycin, 1μg/L rapamycin and blank culture medium for 24 hours respectively. Proliferative capacity was measured by CCK-8 assay, and the migrational ability of the EOC was analysed by scarification test. Results EOC possessed many endothelial characteristics. Immunostaining showed that surface antigens factor Ⅷ, CD34 and Flk-1 were postive. The proliferative capacity and the migrational ability of the EOC were both enhanced after incubation with the 10μ/L SDF-lα （P 〈0. 01）. While 1 μg/L rapamycin not only counteracted such effect （P 〈 0. 01 ）, but also inhibited the biological functions of the EOC （ P 〈 0. 05 ）. Condusoins SDF-lα can activate the proliferative capacity and the migrational ability of EOC. Rapamyc＇in not only counteracts such effect, but also inhibits the biological functions of the EOC directly.