不同氧浓度对骨髓源内皮祖细胞分泌促血管新生相关生长因子的影响

Effect of Different Oxygen Concentration on Neovacularization related Secretion Characteristics of Bone Morrow-derived Endothelial Progenitor Cells

目的:观察培养环境不同氧浓度对骨髓源内皮祖细胞分泌促血管新生相关生长因子的影响。方法:利用密度梯度离心技术分离SD大鼠骨髓单个核细胞,向内皮祖细胞进行诱导分化、扩增、培养和鉴定。然后在不同氧浓度(1%,5%,21%)的环境中培养,于第3天,7天,10天采用酶联免疫吸附试验检测内皮祖细胞分泌血管内皮生长因子(Vascular endothelial growth factor,VEGF)、基质细胞衍生因子-1α(Stromal-derived factor-1α, SDF-1α)、胰岛素样生长因子I(Insulin-like growth factor-I, IGF-I)等生长因子的水平。结果:第3天,不同氧浓度下各组EPC分泌VEGF、SDF-1α、IGF-I无明显差异(P均>0.05)。第7天和第10天,各组EPC分泌IGF-I无明显差异(P均>0.05);但是和21%氧浓度相比,相对低氧浓度(1%和5%)能够明显增强EPC分泌VEGF和SDF-1α(P均<0.001)。结论:适当时间的低氧环境培养能够显著刺激内皮祖细胞分泌VEGF和SDF-1α,进而增强其促血管新生能力。

Objective： This study was to evaluate the effect of different oxygen concentration on EPCs＇ secretion of neovascularization growth factors. Methods： Bone marrow mononuclear cells of SD rat were acquired with density gradient centrifugation method and cultured, induced, and differentiated to EPCs. Then they were cultured respectively under three different oxygen concentration（1 %,5 % and 21 %）. On days 3, 7 and 10, the effect of different oxygen concentration（1 %, 5 % and 21 %） on EPCs＇ secretion（secreting VEGF, SDF-1α and IGF-I） were evaluated by means of ELISA. Results： On days 3, different oxygen concentration（1 %, 5 % and 21 %）did not have effect on EPCs＇ secretion（secreting VEGF, SDF-1α and IGF-I）（all P〉0.05）. On days 7 and 10, there was no difference among three groups for IGF-I（all P〉0.05）. However, compared with 21% oxygen concentration, hypoxia condition（1 % and 5 % oxygen concentration） can stimulate EPCs to secrete VEGF and SDF-1α（P〈0.001）. Conclusions： Appropriate hypoxia condition culture can stimulate EPCs to secrete VEGF and SDF-1α, and enhance their neovascularization function.