姜黄素对内皮祖细胞功能及内皮型一氧化氮合酶的影响

Effects of curcumin on endothelial progenitor cells function and endothelial nitric oxide synthase

目的观察姜黄素对体外培养的人外周血内皮祖细胞(EPCs)数量及功能的影响。方法采用贴壁选择法培养人外周EPCs,培养6 d后,收集贴壁细胞并加入姜黄素(0、5、10、15μmol/L)培养一定时间(6、12、24和48 h)。荧光显微镜鉴定EPCs,分别观察EPCs的体外增殖、黏附、迁移、体外血管生成能力。Griess法检测NO分泌量,并RT-PCR半定量测定eNOS基因表达。结果姜黄素增加外周血EPCs的数量,改善EPCs的体外增殖、黏附、迁移、体外血管生成能力(P<0.01)。并上调eNOS基因表达,促进EPCs来源的NO的释放(P<0.01)。结论姜黄素增加体外EPCs数量及改善其功能,上调eNOS基因表达并增加EPCs来源的NO分泌。

AIM To investigate the effect of curcumin on endothelial progenitor cells （EPCs）. METHODS Total mononuclear cells were isolated from human peripheral blood by ficoll density gradient centrifugation and were cultured. After 6 days of culture in vitro, EPCs were identified as adherent cells double positive for DiI-acLDL/FITC2-UEA- I under a laser scanning confocal microscope and their expression of CD34, CD31 and KDR was detected by flow cytometry. EPCs were then stimulated with curcumin （5, 10, 15 μmol/L） or vehicle control for 48 hours. The proliferation, migration, adhesion activity, vasculogenesis capacity of EPCS and the secretion of NO were assayed. RT-PCR was also performed to determine the eNOS expression after curcumin interference. RESULTS Incubation of human EPCs with curcumin increased the proliferation, adhesion, migration activity and in vitro vasculogenesis capacity （P 〈0.01 ）. Curcumin promoted the secretion of NO（ P 〈 0. 01 ）and upregulated the eNOS expression. CONCLUSION Curcumin promotes the secretion of NO and upregulates the eNOS expression of EPCs. Key words ： curcumin; endothelial progenitor cells ; eNOS