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背景:近年来发现的内皮祖细胞能够促进受损内皮愈合,因而假设支架置入后再狭窄可能与内皮祖细胞的数量和/或活性有关。目的:对比观察冠状动脉支架置入后再狭窄患者与未发生再狭窄患者循环内皮祖细胞数量及活性,验证上述假设。设计、时间及地点:对比观察实验,于2005-03/2007-05在北京世纪坛医院心内科、北京大学第一医院心内科及北京大学医学部生理学与病理生理学系完成。对象:根据复查冠脉造影的结果将来源于北京大学第一医院、既往行冠脉支架置入的患者分为两组:发生支架置入后再狭窄组(n=15)及未发生支架置入后再狭窄的对照组(n=17)。方法:采用密度梯度离心法获取外周血总的单个核细胞,接种到包被有人纤维连接蛋白的24孔培养板,7d后通过激光共聚焦显微镜测定贴壁细胞摄取DiI-acLDL及结合FITC-UEA-Ⅰ的能力,双染色阳性的细胞为正在分化的内皮祖细胞。主要观察指标:外周血单个核细胞培养7d后,通过倒置显微镜计算内皮祖细胞的数量、通过MTT比色法测定内皮祖细胞增殖倍数、通过划痕试验定性观察内皮祖细胞的迁移数量以及通过黏附试验测定内皮祖细胞的黏附率。结果:再狭窄组内皮祖细胞数量明显低于对照组(P=0.001),再狭窄组的内皮祖细胞增殖倍数明显低于对照组(P<0.05),再狭窄组的内皮祖细胞迁移数量明显低于对照组,但两组之间的内皮祖细胞黏附率差异无显著性意义(P>0.05)。结论:冠状动脉支架置入后再狭窄患者的内皮祖细胞数量及增殖能力、迁移能力明显下降,说明支架置入后再狭窄可能与内皮祖细胞的数量和/或活性有关。
BACKGROUND: Endothelial progenitor cells (EPCs) found in recent years are able to promote damaged endothelial healing. Therefore, it is hypothesized that restenosis after stent implantation may be related to the number and / or activity of endothelial progenitor cells. OBJECTIVE: To compare and contrast the number and activity of circulating endothelial progenitor cells in patients with restenosis after coronary stent implantation and without restenosis, and to verify the above hypothesis. DESIGN, TIME AND SETTING: Comparative observation experiment was performed at Department of Cardiology, Beijing Shijitan Hospital, Department of Cardiology, Peking University First Hospital, and Department of Physiology and Pathophysiology, Peking University from March 2005 to May 2007. Subjects: According to the results of the review of coronary angiography, patients from the First Hospital of Peking University, who had undergone coronary stenting, were divided into two groups: group with restenosis after stent placement (n = 15), and stent placement In the later restenosis control group (n = 17). Methods: Peripheral blood mononuclear cells were obtained by density gradient centrifugation and inoculated into 24-well plates coated with human fibronectin. The expression of DiI-acLDL and the binding of FITC-UEA to adherent cells were measured by laser confocal microscopy 7 days later -I ability, dual-positive cells are differentiation of endothelial progenitor cells. MAIN OUTCOME MEASURES: Peripheral blood mononuclear cells were cultured for 7 days, the number of endothelial progenitor cells was calculated by inverted microscope, the multiplication rate of endothelial progenitor cells was determined by MTT colorimetric assay, the number of migration of endothelial progenitor cells was observed qualitatively by scratch test, The assay determines the adhesion rate of endothelial progenitor cells. Results: The number of endothelial progenitor cells in restenosis group was significantly lower than that in control group (P = 0.001). The proliferation of endothelial progenitor cells in restenosis group was significantly lower than that in control group (P <0.05) Lower than the control group, but there was no significant difference in the adhesion rate of endothelial progenitor cells between the two groups (P> 0.05). CONCLUSION: The number of EPCs, the proliferative capacity and migration ability of patients with restenosis after coronary stenting are significantly decreased, suggesting that restenosis may be related to the number and / or activity of endothelial progenitor cells after stent implantation.