重组人促红细胞生成素与骨髓间充质干细胞的迁移及黏附☆

背景:促红细胞生成素可促进内皮祖细胞的增殖分化并增强其黏附性。目的:观察重组人促红细胞生成素干预对人骨髓间充质干细胞迁移和黏附能力及相关细胞信号传导通路的影响。方法:体外培养人骨髓间充质干细胞,使用重组人促红细胞生成素干预第6代人骨髓间充质干细胞。分别用PI3K/Akt通路特异抑制剂Ly294002或p38MAPK通路特异激动剂anisomycin或ERK1/2通路特异抑制剂U0126预处理。结果与结论:重组人促红细胞生成素可使人骨髓间充质干细胞的PI3K/Akt通路磷酸化水平升高,抑制p38MAPK通路磷酸化水平,对人骨髓间充质干细胞的ERK通路和总Akt、总p38MAPK水平无明显影响。重组人促红细胞生成素作用组中迁移细胞数目显著高于对照组(P<0.01),黏附细胞数亦明显增加(P<0.01),PI3K/AKT通路特异抑制剂Ly294002预处理后重组人促红细胞生成素对迁移能力的作用消失,p38MAPK通路特异激动剂anisomycin预处理对两种作用影响均不明显。提示重组人促红细胞生成素具有增强人骨髓间充质干细胞迁移和黏附能力的作用,其增强迁移能力的作用与激活人骨髓间充质干细胞的PI3K/Akt通路有关,但是它对黏附能力的作用与PI3K/Akt和p38MAPK通路均无关。

Effects of recombinant human erythropoietin on migration and adhesion of human mesenchymal stem cells

BACKGROUND：Previous studies have demonstrated that erythropoietin can promote the proliferation, differentiation and adhesion of endothelial progenitor cells. OBJECTIVE：To investigate the effects and related cell signaling pathways of recombinant human erythropoietin on migration and adhesion of human mesenchymal stem cells. METHODS：After being cultured in vitro, passage 6 human mesenchymal stem cells were treated with recombinant human erythropoietin. Recombinant human erythropoietin, phosphorylation of ERK 1/2, p38MAPK and PI3K/Akt were detected by western blot. After 24 hours of pretreatment with 1U/mL recombinant human erythropoietin, cell migration assay was performed in Transwell chambers, and adhesion assay was performed by plastic dishes. RESULTS AND CONCLUSION：Recombinant human erythropoietin could increase phosphorylation of PI3K/Akt pathway of human MSCs, but reduce phosphorylation of p38MAPK. It had no obvious effect on ERK1/2 pathway and total proteins of Akt and p38MAPK. The number of cells crossing the filter was significantly increased in the recombinant human erythropoietin group （P 0.01）. This effect was completely inhibited by concomitant incubation with Ly294002. After treated with recombinant human erythropoietin, adherent cells increased significantly （P 0.01）. Neither Ly294002 nor anisomycin could inhibit this effect. Recombinant human erythropoietin could increase migratory and adhesive capacities of human mesenchymal stem cells. Activation of PI3K/Akt pathway was involved in the effect of recombinant human erythropoietin on cell migration. The effect of rhEPO on adhesion capacity had nothing to do with PI3K/Akt and p38MAPK pathway.