【作者】 封灏；

【导师】 刘建民； 许奕； 洪波； 赵文元； 黄清海；

【作者基本信息】 第二军医大学 ， 外科学， 2011， 硕士

【摘要】 目的研究未破裂颅内动脉瘤与外周血中内皮祖细胞(endothelial progenitor cells,EPCs)数量的关系。方法选择未破裂颅内动脉瘤患者24例和正常对照16例,用密度梯度离心法从外周血获取单个核细胞,接种在人纤维连接蛋白覆盖的培养瓶,经加入血管内皮生长因子165和碱性成纤维细胞生长因子的EGM-2MV培养基诱导培养。动态观察其增殖过程。对培养14天后贴壁的EPCs细的进行计数、鉴定和分析。激光共聚焦显微镜下吞噬FITC标记的荆豆凝集素和结合Dil标记的acLDL双染色阳性细胞为成熟的EPCs；并用流式细胞仪检测细胞表面CD34、CD133和VEGFR-2抗原,测定细胞表面抗原CD34、CD133和VEGFR-2三抗阳性的细胞数以及比例。分析比较两组外周血中内皮祖细胞的数量差异,并在病例组进行相关因素研究,分析动脉瘤最大径同外周血EPCs细胞数关系。结果一、EPCs的数量检测中：颅内动脉瘤患者外周血EPCs数量较对照组明显减少(5.1±1.5)×104比(43.3±29.8)×104, (P<0.05),流式细胞仪分析结果显示CD34+/CD133+/VEGFR-2+的细胞数比例(6.94%±1.78%比17.83%±3.41%,P<0.05)有显著差异。二、基本资料中：颅内动脉瘤患者甘油三酯(1.86±1.37) mmol/L比健康对照组(1.58±0.98) mmol/L明显升高(P<0.05)；颅内动脉瘤患者胆固醇(3.85±1.67) mmol/L比健康对照组(3.67±1.26) mmol/L明显升高(P<0.05)；颅内动脉瘤患者HDL (1.08±0.26) mmol/L比健康对照组(1.96±0.64)mmol/L显著降低(P<0.05)。三、24例病例组动脉瘤最大径(5.4±3.4)mm。病例组的动脉瘤最大径与病例组EPCs计数无统计相关性(P>0.05)结论未破裂颅内动脉瘤患者外周血EPCs的数量与健康人比明显降低。EPCs在未破裂颅内动脉瘤中的作用值得进一步研究探讨。更多还原

【Abstract】 Objective:To investigate the morphology, growth and counts of endothelial progenitor cells in peripheral blood of patients with unruptured intracranial aneurysms(UIAs). Method: Twenty-four patients of UIA Group and 16 of Control Group were analysed. Mononuclear cells from peripheral blood were harvested by density gradient centrifugation. Then the cells were seeded onto a bottle which was precoated with human fibronectin and cultured in medium EGM-2MV supplemented with vascular endothelial growth factor-165 and basic fibroblast growth factor. The proliferation course of EPCs was observed. After fourteen-day culturing, the adherent cells were identified through morphology observation and the colony count was performed under microscope. The biological functions of adherent cells were examined with laser confocal microscope. The double dyeing positivity, that is adsorption of Ulex europaeus agglutinin-1 (UEA-1) labeled by fluorescein isothiocyanate (FITC) and Dil-acLDL internalization, were considered as EPCs. Flow cytometry was used to count the EPCs by identifying three cell surface markers:CD34, CD 133 and VEGFR-2. The number of circulating endothelial progenitor cells between the two groups was compared, and the correlation of the maximum aneurysm diameter and the number of circulating EPCs was analyzed.Results:1、The counts of endothelial progenitor cells in peripheral blood were significantly lower in patientswith UIAs [(5.1±1.5)×104] than that in control group [(43.3±29.8)×104] (P<0.05). Moreover, the portion of CD34+/CD133+/VEGFR-2+cells were significantly decreased in patients with UIAs than that in controls (6.94%±1.78% vs 17.83%±3.41%,P<0.05).2, The value of triglycerides in patients with UIA [(1.86±1.37) mmol/L] was significantly higher than that of control group [(1.58±0.98) mmol/L], (P<0.05); The cholesterol is significantly higher in UIA [(3.85±1.67) mmol/L] than that of healthy control group [(3.67±1.26) mmol/L],(P<0.05); HDL significantly less in intracranial aneurysms is [(1.08±0.26) mmol/L] than that of healthy control group [(1.96±0.64) mmol/L], (P<0.05).3、A total of 24 patients were recruited in UIAs group, the maximum aneurysm diameters were (5.4±3.4) mm. No significant correlation between the maximum aneurysm diameters and EPCs counts in patient group has been shown (P> 0.05)Conclusion:The results shew that the number of EPCs in UIAs group was significantly lower than that of control group. Further study on the role of EPCs in UIAs may be promising in future.更多还原