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作 者:韩国雄[1] 李建平[1] 严文琪[1] 张富财[1] 蒋白丽 彭海[1] 高云梅[1] 解友邦[1] 李文倩[1] 阿祥仁[1] 冯建明[1]
机构地区:[1]青海省人民医院,810007
出 处:《高原医学杂志》2016年第4期12-16,共5页Journal of High Altitude Medicine
摘 要: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 study the effects of hypoxia on the expression of hypoxia-inducible factor 1(HIF-1)and the production of pro-angiogenic factors by bone marrow mesenchymal stem cells(BMMSCs).Methods:BM- MSCs were isolated,cultured and identified using flow cytometry.BM- MSCs were induced to differentiate into adipocytes and osteoblasts.Identical BM-MSCs wererespectively cultured in normoxia(20% oxygen)and hypoxia(1% oxygen)to compare the expression of HIFs and the production of vascular growth factors.The mRNA expression of HIFs was detected using quantitative real-time PCR.The concentrations of vascular growth factors in the culture medium of BM-MSCs were determined using enzyme-linked immunoassay kits.Results:The expression of HIF-1αand HIF-1βmRNA in hypoxia conditions were higher than those of normoxic conditions(P0.05).The concentrations of VEGF in hypoxia conditions was higher than that of normoxic conditions(P=0.000).However,the concentrations of Angiogenin and TGF-βin hypoxia conditions were lower than those of normoxic conditions(P0.05)while there was no significant difference in Angiopoietin-2and IL-6between two groups.Conclusion:ypoxia can promote the expression of HIF-1of BM-MSCs and influence on the secretion of vascular growth factors simultaneously.
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