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目的研究前凋亡因子Bim在内皮祖细胞(EPCs)自然凋亡中的作用。方法采集健康产妇产后2h脐静脉血,6%羟乙基淀粉沉淀及密度梯度离心法提取脐血中单个核细胞,培养10d,免疫荧光法及免疫组化法鉴定EPCs。采用流式细胞仪检测EPCs凋亡率,RT-PCR分析不同时间点(0,6,12,24,48h)BimmRNA的表达。结果体外培养的EPCs随培养时间发生自然凋亡,各时间点细胞凋亡百分比较对照组(0h)高;RT-PCR显示在一定时间内前凋亡因子BimmRNA随培养时间延长表达增高。结论 Bim参与了EPCs自然凋亡的过程,是重要的促细胞凋亡因子。
Objective To study the role of pro-apoptotic factor Bim in natural apoptosis of endothelial progenitor cells (EPCs). Methods Umbilical cord blood, 6% hydroxyethyl starch pellets and density gradient centrifugation were used to collect mononuclear cells of umbilical cord blood from healthy pregnant women at 2 hours postpartum. The cells were cultured for 10 days. EPCs were identified by immunofluorescence and immunohistochemistry. The apoptotic rate of EPCs was detected by flow cytometry. The expression of Bim mRNA was detected by RT-PCR at different time points (0, 6, 12, 24, 48h). Results EPCs cultured in vitro were apoptosis induced by culture time. The percentage of apoptotic cells at each time point was higher than that of the control group (0 h). The expression of Bim mRNA increased with the prolongation of culture time by RT-PCR. Conclusion Bim is involved in the natural apoptosis of EPCs and is an important pro-apoptotic factor.