氯化高铁血红素对人晚期内皮祖细胞数量和血红素加氧酶-1表达的影响

Effects of hemin on number of human late endothelial progenitor cells and expression of heme oxygenase-1

目的观察氯化高铁血红素(简称血红素)在体外对人晚期内皮祖细胞(EPCs)数量和血红素加氧酶-1(HO-1)的影响。方法从健康分娩产妇脐带静脉血中分离提取单个核细胞,体外培养诱导分化为晚期EPCsO在不同体积分数胎牛血清(FBS)条件下用血红素(10μmol/L)干预并设置对照组,干预24h后,锥虫蓝染色计数,细胞计数试剂盒-8(CCK-8)检测细胞毒性,Western印迹法检测HO-1的表达。结果在低体积分数(0.2%)FBS条件下,与对照组相比,血红素减少了晚期EPCs的数量,但随着FBS体积分数的逐渐提高,血红素对晚期EPCs增殖的抑制能力减弱,促增殖能力逐渐增强(P<0.05),且诱导HO-1的表达逐渐提高。结论细胞体外培养中,FBS体积分数在血红素对晚期EPCs数量和活性的作用方面有着重要的影响,且在高体积分数(5%)FBS条件下,血红素促进细胞增殖的效应初步推测是通过诱导HO-1高表达来实现的,但其具体机制和其中涉及的信号转导通路有待进一步探讨。

Objective To investigate the effects of Hemin on the number of late endothelial progenitor cells（EPCs） and the expression of heme oxygenase-1. Methods Mononuclear cells （MNCs） were isolated from the umbilical cord blood of healthy women after delivery and were induced to differentiate into late EPCs in vitro. Several groups of attached late EPCs were plated in the absence or presence of Hemin （ 10 μmol/L） for 24 h with different concentrations of FBS. Then cells were counted using a cell counter with trypan blue, and cell proliferation was evaluated using the cell countion kit-8 （CCK-8） assay kit. The expression of heme oxygenase-1 （HO-1） was detected by Western blotting analysis. Results Heroin decreased the number of late EPCs in presence of low concentration of FBS compared with control group, but this inhibitory effect on late EPCs was reversed when the FBS concentration increased to 5%, under which condition hemin improved late EPCs proliferation （P〈0.05） and the expression of HO-1. Conclusion Late EPCs show a biphasic response to hemin under different concentrations of FBS, and it can increase late EPCs proliferation in presence of high concentration of FBS, which might be related to the induced expression of HO-1.