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目的:探讨低氧对骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMMSCs)中缺氧诱导因子-1(hypoxia inducible factor-1,HIF)基因表达水平及其分泌的血管生长因子的影响。方法:分别分离、培养来源于7例正常人骨髓的间充质干细胞,P3代时进行形态学、免疫表型及多向分化实验鉴定。低氧(1%)及常氧(20%)条件下培养骨髓间充质干细胞,Real-time PCR方法检测骨髓间充质干细胞HIF系统(HIF-1α、HIF-1β)基因表达差异,ELISA方法检测分析骨髓间充质干细胞分泌的血管生长因子(VEGF、Angiogenin、Angiopoietin-2、TGF-β和IL-6)浓度。结果:低氧条件下HIF-1α和HIF-1β基因表达水平均增高(P<0.05),骨髓间充质干细胞分泌的VEGF水平高于常氧培养组(P=0.000),Angiogenin和TGF-β水平低于常氧培养组(P<0.05),而Angiopoietin-2和IL-6无显著性差异(P>0.05)。结论:低氧促进骨髓间充质干细胞HIF-1的表达的同时,对其分泌的血管生长因子产生不同影响。
Objective: To investigate the effect of hypoxia on hypoxia inducible factor-1 (HIF) gene expression and its secretion of vascular growth factor in bone marrow mesenchymal stem cells (BMMSCs). Methods: Mesenchymal stem cells derived from bone marrow of seven normal individuals were isolated and cultured respectively. Morphological, immunophenotypic and multidirectional differentiated experiments were performed on P3 generation. BMSCs were cultured under hypoxia (1%) and normoxia (20%). The expression of HIF-1α and HIF-1β in bone marrow mesenchymal stem cells was detected by Real-time PCR. The concentrations of VEGF, Angiogenin, Angiopoietin-2, TGF-β and IL-6 secreted by bone marrow mesenchymal stem cells were detected and analyzed. Results: The expression of HIF-1α and HIF-1β in hypoxia increased (P <0.05), the level of VEGF secreted by bone marrow mesenchymal stem cells was higher than that in normoxia (P = 0.000), Angiogenin and TGF-β The levels of Angiopoietin-2 and IL-6 were not significantly different (P> 0.05). CONCLUSION: Hypoxia can promote the expression of HIF-1 in bone marrow mesenchymal stem cells, and at the same time, it has different effects on the secretion of vascular endothelial growth factor.