CAJ | 학술논문

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.
목적 체외관찰불동농도록화고철혈홍소(hemin)대인제혈래원적만기내피조세포(late EPCs)수량화조망적영향.방법 용밀도제도리심법분리제취인제정맥혈래원적단핵세포,재체외유도분화위late EPCs.생장상태량호적제2～3대late EPCs수궤접수불동농도hemin(0、5、10、15、20 μmol/L)간예24 h후계속배양,태반람염색법검측세포활성,활세포계수시제합검측세포증식정황,점부시험측정관찰세포점부능력,류식세포의검측세포조망.결과 여대조조상비,교저농도hemin(5μmol/L혹10 μmol/L)가현저촉진late EPCs적활성;당hemin농도위5、10혹15 μmol/L시,late EPCs증식급점부작용명현,이조망수도억제.기중hemin농도위10 μmol/L시저충작용최강,이고농도hemin(20μmol/L)칙표현위상반적작용효과.차외,hemin촉증식、점부급억제조망적작용구유시간의뢰성,재24 h작용최강.결론 hemin재저농도시현저증가료late EPCs적수량、촉진점부、억제조망,재고농도시칙기상반작용.
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.