目的：探讨MSCs在缺氧环境中细胞生物学特性及相关生长因子的变化，为修复组织损伤提供临床应用依据。方法体外进行MSCs缺氧处理，免疫荧光染色观察细胞特性，RT-PCR检测VEGF、bFGF、TGF-β和IL-1α基因，从细胞分化、生长因子和细胞信号通路等方面，来探讨MSCs在缺氧环境下发生的转变。结果 MSCs经1%O2缺氧培养24 h后，镜下观察示细胞成活良好，未见形态、生长方式等发生明显变化。免疫荧光及双荧光染色观察缺氧处理24 h的MSCs有向内皮细胞，特别是血管内皮转化的趋势。 VEGF、bFGF、TGF-β和IL-1α在缺氧处理后的MSCs中表达均增强，而未经缺氧处理的MSCs相关因子表达微弱。结论在缺氧的环境下，MSCs可向血管内皮细胞方向分化，并且与促进血管形成和生长相关的VEGF、bFGF、TGF-β和IL-1α基因表达均增强，可直接或间接促进损伤组织的血管化。%Objective To investigate the biological characteristics and related factors of mesenchymal stem cells (MSCs) under hypoxia and to extend the knowledge for repairing clinical tissue injury. Methods MSCs were cultured and exposed to the hypoxic condition in vitro. The cell properties were observed by immunofluorescence staining, RT-PCR were performed to investigate the expression of VEGF、bFGF、TGF-β and IL-1α, in order to explore the changes of MSCs under hypoxia from the aspects of cell differentiation, related factors and cell signaling pathways. Results Cultured under hypoxia for 24 hours, cells were observed under inverted microscope and survived well, and no significant changes of cell morphology and growth patterns were observed. Immunofluorescence and double immunofluorescence staining found out there was a tendency that MSCs cultured in hypoxia transform into endothelial cells, particularly vascular endothelial cells. The expression of VEGF, bFGF, TGF-βand IL-1αwere all up-regulated in MSCs exposed to the hypoxic condition, but a weak expression in MSCs cultured in normoxia. Conclusion Under the hypoxic condition, MSCs can differentiate into vascular endothelial cell, and expression of factors VEGF, bFGF, TGF-β and IL-1α, which can enhance the formation and growth of vascular, are all up-regulated, directly or indirectly accelerate the vascularization in tissue injury.
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