一氧化碳对人外周血内皮祖细胞功能及NO生成的影响

Effects of Carbon Monoxide on Function and NO Production of Peripheral Bloodderived Endothelial Progenitor Cells in Vitro

目的:研究一氧化碳(CO)对人外周血内皮祖细胞(EPCs)增殖、迁移、黏附功能及分泌NO的影响。方法:密度梯度离心法获取人外周血单个核细胞,接种于预先包被人纤维连接蛋白的培养板上,培养1周后鉴定EPCs。将培养1周后的EPCs分为对照组和干预组,干预组分别加入25μM、50μM、100μM、200μM的一氧化碳释放分子-3(CORM-3)培养24 h。测定EPCs的增殖、迁移、黏附和分泌NO能力。结果:与对照组比较,干预组EPCs增殖、迁移、黏附能力增强,培养上清液中NO浓度增加(P<0.05),且在100μM时作用最显著。结论:CO可促进体外培养的EPCs增殖、迁移、黏附和生成NO,提示CO可能通过改善EPCs功能加速血管再内皮化和促进血管新生。

Objective： To investigate the effects of carbon monoxide （CO）on the proliferation, migration, adhesion and nitric oxide（NO） production of endothelial progenitor cells（EPCs） from human peripheral blood in vitro. Method： Total mononuclear cells were isolated from human peripheral blood by ficoll density gradient centrifugation and cultured on fibronectin-coated six-well plates. After 7 days , EPCs were identified with dual fluorescence staining and flow cytometry. EPCs were divided into control group and intervention group at the seventh day, and 25 μM, 50μM, 100 μM, 200 μM carbon monoxide releasing molecule-3 （CORM-3） were added in the intervention group, respectively. The proliferative, migratory and adhesive abilities and NO production were observed after 24 h. Results：Compared with control group , the proliferative , migratory and adhesive activities of EPCs were enhanced by CORM-3 , and the NO concentration in the culture supernatant were higher than the control group （P ＆lt; 0.05）. In addition, CORM-3 had the most obvious effect on EPCs at 100μM level. Conclusion： CO could improve the proliferation, migration , adhesion capacity and NO production of EPCs in vitro , which suggests that CO could possibly play an role in accelerating re-endothelialization and promoting angiogenesis by improving the function of EPCs.