摘要
目的 体外观察不同浓度氯化高铁血红素（hemin）对人脐血来源的晚期内皮祖细胞（late EPCs）数量和凋亡的影响.方法 用密度梯度离心法分离提取人脐静脉血来源的单核细胞,在体外诱导分化为late EPCs.生长状态良好的第2～3代late EPCs随机接受不同浓度hemin（0、5、10、15、20 μmol/L）干预24 h后继续培养,台盼蓝染色法检测细胞活性,活细胞计数试剂盒检测细胞增殖情况,粘附试验测定观察细胞粘附能力,流式细胞仪检测细胞凋亡.结果 与对照组相比,较低浓度hemin（5μmol/L或10 μmol/L）可显著促进late EPCs的活性;当hemin浓度为5、10或15 μmol/L时,late EPCs增殖及粘附作用明显,而凋亡受到抑制.其中hemin浓度为10 μmol/L时这种作用最强,而高浓度hemin（20μmol/L）则表现为相反的作用效果.此外,hemin促增殖、粘附及抑制凋亡的作用具有时间依赖性,在24 h作用最强.结论 hemin在低浓度时显著增加了late EPCs的数量、促进粘附、抑制凋亡,在高浓度时则起相反作用.
Objective To investigate the effects of hemin on the quantity and apoptosis of human umbilical cord blood-derived late endothelial progenitor cells （EPCs） in vitro.Methods Mononuclear cells were isolated from human cord blood by density gradient centrifugation and were induced to differentiate to late EPCs in vitro.The second to third generation of attached late EPCs in good state were randomly plated for 24 h under different concentrations（0, 5, 10, 15 and 20 μmol/L）of hemin.Cell viability and proliferation were measured with typan blue staining and cell counting kit-8, respectively.Cell adhesion was analyzed by adhesive assay, and cell apoptosis was detected by flow cytometry.Results Compared to control group, hemin promoted viability of late EPCsat lower concentrations（5and 10μmol/L）.Meanwhile, proliferation and adhesion were also improved and apoptosis was inhibited when the concentrations of hemin were 5,10, or 15 μ mol/L.All these effects were most prominent when hemin concentration was 10 μmoL/L, while the effects above were reversed when hemin concentration was moderated to 20 μmol/L.In addition, hemin showed a time-dependent manner in promoting cell proliferation and adhesion, and inhibiting apoptosis.That effects were most obvious at 24 h.Conclusions Lower concentration of herin augments the quantity and adhesion of late EPCs, inhibits cell apoptosis, while higher concentration present the reversed effects.
出处
《中国医师杂志》
CAS
2015年第10期1520-1523,共4页
Journal of Chinese Physician
