摘要
目的:观察不同浓度血管内皮生长因子(VEGF)对体外培养人外周血内皮祖细胞(EPCs)生物学功能的影响,探讨VEGF促进EPCs分裂生长的合适浓度。方法:密度梯度离心法获取人外周血单个核细胞(MNCs),接种至人纤维连接蛋白(HFN)包被的培养板上,培养4 d后收集贴壁细胞,换用配有不同浓度(对照组、10μg/L、20μg/L、50μg/L)VEGF的培养基继续培养3 d后进行细胞免疫组化鉴定EPCs,采用MTT比色法、改良的Boyden小室和黏附能力测定实验,观察EPCs的增殖、迁移和黏附能力。结果:从人外周血能成功分离EPCs细胞,并能分化为血管内皮细胞;从冠心病患者分离的EPCs增殖能力较非冠心病患者弱;VEGF在较低浓度(10μg/L和20μg/L)时即能显著促进EPCs生长的各项生物学指标,但高浓度(50μg/L)时并不能进一步增强这一效应;低浓度(10μg/L)下VEGF对冠心病患者作用较非冠心病患者弱,高浓度时对两者促进作用相近。结论:冠心病患者EPCs功能显著减弱,较低浓度的VEGF即可显著增强EPCs的各项生物学功能,可能对损伤血管的再内皮化有益。
AIM： To investigate the effect of vascular endothelial growth factor （VEGF） on the biological function of peripheral endothelial progenitor cells （EPCs） and to find the suitable concentration to promote the growth of EPCs. METHODS ： Total mononuclear cells （MNCs） were isolated from peripheral blood by density gradient centrifugation, and then the cells were plated on fibronectin - coated culture dishes. After culture for 4 days, attached cells were incubated with VEGF in a series of concentrations （0, 10, 20 and 50μg/L） for72 h, then attached cells were characterized with immunohistochemistry. EPC proliferation and migration activity were assayed with MTT assay and modified Boyden chamber assay, respectively. EPC adhesion assay was performed by replating MNCs on fibronectin - coated dishes, and then the adherent cells were counted. RESULTS： The EPCs from MNCs were successfully isolated and were differentiated to endothelial cells （ECs）. Incubation of isolated human MNCs with VEGF increased the proliferative, migratory and adhesive capacities of EPCs, and this effect was most prominent when the concentration of VEGF was 20 μg/L after 72 hours. At the same concentration of VEGF, the functions of EPCs from patients with masculine coronary arteriography were lower than those of EPCs from patients with negative coronary arteriography. CONCLUSION： Functional activities of EPCs are decreased in patients with masculine coronary arteriography. The results suggest that the low concentration of VEGF may improve functional activities of EPCs.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2007年第10期1896-1899,共4页
Chinese Journal of Pathophysiology
基金
浙江省科学技术基金资助项目(No.2004C33045)
关键词
内皮生长因子
内皮祖细胞
冠状动脉疾病
Endothelial growth factors
Endothelial progenitor cells
Coronary disease
