心脑血管病防治2016，Vol.16Issue(2)：86-90,5.DOI:10.3969/j.issn.1009-816x.2016.02.03
5_氮杂_2’_脱氧胞苷对内皮祖细胞分化的影响及机制研究
Effects and Mechanism of 5_aza_2’_deoxycytidine on the Differentiation of Endothelial Progenitor Cells
摘要
Abstract
Objective To investigate the effects and mechanism of 5_aza_2’_deoxycytidine induce endothelial progenitor cells to dif-ferentiate into endothelial cells.Methods mononuclear cells were separated from umbilical cord blood ,using Ficoll_Paque lymphocyte separation medium and 6% hydroxyethyl starch sedimentation ,which were cultured and differentiate into endothelial progenitor cells (EPCs).The EPCs were identified using immunohistochemistry and inverted phase contrast microscope.After 7 days ,1μmol/L (ie dC1 group) or 3μmol/L (ie dC2 group) of 5_Aza_dC was added.0.0076% DMSO was used as intervention in the negative group.CD34+ ,CD31+ expression influenced by 5_Aza_dC Treatment after 0 ,3 ,7 ,14d were evaluated by flow cytometry ,VE_cadherin , Tie_2 ,gene expression of vWF were analyzed by RT_PCR ,protein expression of vWF was tested by western blotting ,gene expression of CpG island methylation of BMP4 was detected by MSP. Results ① monocytes were proved to differentiate into EPCs by inverted phase contrast microscope ,VE_cadherin ,Tie_2 ,vWF ,KDR ,CD34 and CD133 and other factors ;② It was showed in flow cytome-try that CD34+ (respectively ,30.2% ,6.7% ) in dC2 group by 5_Aza_dC at 3d ,7d was statistical significantly higher than negative group (46.7% ,40.8% ;all P<0.05) ,CD31+ (47.9% ) in dC2 group by 5_Aza_dC at 7d was statistical significantly higher than negative group (31.3% ;all P<0.05) ③It was showed in RT_PCR that Tie_2 ,VE_cadherin gene in dC2 group at 14d were significantly higher than negative group ( t=8.12 ,12.28 ,all P<0.05 );Tie_2 ,vWF ,VE_cadherin gene in dC1 ,dC2 group at 3d ,7d ,14d t were higher than at 0d (all P<0.05) ;④In Western blotting anal-ysis ,it was showed that vWF protein expression in dC2 group at 0d , 3d ,7d were higher than negative group in those days ( P<0.05) , vWF protein expression in dC1 ,dC2 group at 3d ,7d were higher than the protein expression at 0d in each two group (all P<0.05);⑤After methylated treatment ,the expression of BMP4 in dC1 ,dC2 group was decreased ,while during unmethylated treatment , BMP4 in dC1 ,dC2 group was induced to increased expression .Conclusions 5_Aza_dC could rapidly induce endothelial progenitor cells to differentiate into endothelial cells ,whose mechanism may connected with increased expression of the differentiation genes BMP4 ,and induced expression of Tie_2 ,VE_cadherin and vWF.关键词
5_氮杂_2’_脱氧胞苷/内皮祖细胞/分化/内皮细胞Key words
5_aza_2’_deoxycytidine/Endothelial progenitor cells/Differentiation/Endothelial cells分类
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刘加远,杨国源,杨志军,曲美洁,戴宜武,徐如祥..5_氮杂_2’_脱氧胞苷对内皮祖细胞分化的影响及机制研究[J].心脑血管病防治,2016,16(2):86-90,5.基金项目
国家自然科学基金 （）
