糖基化终末产物对人外周血内皮祖细胞数量及功能的影响  

Effects of AGEs on number and function of EPCs from human peripheral blood

作　　者：闫醒军[1] 边忠平[1] 刘勇[2] 黄启荣[3] 崔驰[4] 周秀娟[5] 钟武[2] 施森[2] 何延政[2] 

机构地区：[1]唐山市工人医院心脏血管外二科,唐山063000 [2]泸州医学院附属医院血管外科 [3]四川省邛崃市人民医院普外科 [4]四川省绵阳市中心医院普外科 [5]四川省雅安市人民医院普外科

出　　处：《中华老年心脑血管病杂志》2009年第12期972-975,共4页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

摘　　要：目的观察糖基化终末产物(AGEs)对体外培养的人外周血内皮祖细胞(EPCs)数量及功能的影响。方法体外培养的人外周血EPCs根据不同AGEs-人血白蛋白(HSA)浓度分为对照纽、正常组(终浓度7.5 mg/L)、干预1组(终浓度1 5 mg/L)干预2组(终浓度30 mg/L)和干预3组(终浓度60 mg/L)。采用密度梯度离心法获取人外周血单个核细胞;激光共聚焦显微镜检测其对FITC标记荆豆凝集素-1的吸附和Dil-标记乙酰化低密度脂蛋白进行细胞功能鉴定;加入AGEs后,分别采用MTT法、Boyden小室测定EPCs的增殖、迁移能力;人纤维连接蛋白检测EPCs的黏附能力;用体外血管生成分析试剂盒检测不同浓度AGEs HSA作用后的EPCs成血管能力。结果高浓度AGEs可减少EPCs贴壁细胞数量(P<0.01),减弱EPCs的黏附(P<0.01)、增殖(P<0.01)、迁移(P<0.05)和成血管能力(P<0.01)。结论 AGEs可减少人外周血EPCs的数量并使用其功能受损。Objective To study the effect of advanced glycation end products（AGEs） on number and biological characteristics of endothelial progenitor cells（EPCs） from cultured human peripheral blood. Methods Mononuclear cells（MNCs） isolated from human peripheral blood by density gradient centrifugation combined with adherent cell filtration were cultured. The biological functions of adherence cells were examined by the adsorption of Ulex europaeus agglutinin-1（UEA-1） labeled by fluorescein isothiocyanate（FITC） and Dil-acLDL internalization on 4th day during culture. The expression of specific antigens（CD34, CD133） on cell surface was analyzed by cytometer. After 4 days,adherent cells were cultured with different concentrations of AGEs for 48 hours. The MTT assay and Boyden chamber were used to observe the proliferation and migration of EPCs,human fibronectin was used to examine adhesion ability of EPCs, in vitro angiogenesis assay kit was used to observe angiogenesis of EPCs which had been exposed to different concentrations of AGEs HSA. Results AGEs dose-dependently decreased the number of EPCs （P〈0. 01）,weakened EPCs proliferation （P 〈 0.01）, migration （P 〈 0. 01）, adhesive capacity （P 〈0.05）, and increased the apoptosis rate of EPCs in the early stage （P 〈 0.01）. Conclusion AGEs can decrease the number of EPCs from human peripheral blood and induce its dysfunction.

关 键 词：糖基化终产物 高级 内皮细胞 流式细胞术 单核细胞 造血干细胞 细胞粘附分子 

分 类 号：R363[医药卫生—病理学]