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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 observe the effect of advanced glycation end products (AGEs) on the number and function of EPCs in vitro. Methods EPCs from human peripheral blood were divided into control group, normal group (final concentration 7.5 mg / L) and intervention group 1 (final concentration 15 mg / L) according to the concentration of AGEs-human serum albumin (HSA) Group (final concentration 30 mg / L) and intervention group 3 (final concentration 60 mg / L). Human peripheral blood mononuclear cells were obtained by density gradient centrifugation. The laser scanning confocal microscope was used to detect the adsorption of FITC-labeled lepidin-1 and Dil-labeled acetylated low density lipoprotein (LDL). After addition of AGEs, The proliferation and migration of EPCs were measured by Boyden chamber by MTT assay. The adhesion ability of EPCs was detected by human fibronectin. The angiogenesis ability of EPCs treated with different concentrations of AGEs was detected by in vitro angiogenesis assay kit. Results High concentrations of AGEs decreased the number of adherent cells (P <0.01), decreased EPCs adhesion (P <0.01), proliferated (P <0.01), migrated (P <0.05) and angiogenic ability (P <0.01). Conclusion AGEs can reduce the number of EPCs in human peripheral blood and use their impaired function.