摘要
目的探讨酸性成纤维细胞生长因子(aFGF)对人脐带血中内皮祖细胞凋亡的影响及机制。方法选择健康产妇脐带血,密度梯度离心法获取脐带血单个核细胞,培养7天后,收集贴壁细胞并加入不同浓度(2、5、10μg/L)aFGF干预24 h,流式细胞仪检测aFGF对细胞凋亡的影响,逆转录-聚合酶链反应检测Bcl-2 mRNA表达,Western Blot检测Bcl-2蛋白表达。同时对作用效果最为显著的组(5μg/L aFGF组)分别培养6、12、24、48 h,分别观察内皮祖细胞数量及凋亡状况,从而对其时效关系进行观察。结果与对照组比较,不同浓度的aFGF可以显著抑制内皮祖细胞凋亡(P<0.05);5μg/L aFGF作用24 h时对细胞凋亡抑制的影响最为显著(P<0.05)。Bcl-2mRNA和蛋白的表达显著高于对照组。结论 aFGF能抑制人脐带血内皮祖细胞的凋亡,并通过上调凋亡抑制基因Bcl-2而发挥抑制细胞凋亡的作用。
Aim To explore the effect of acid fibroblast growth factor （aFGF） on the apoptosis in endothelial progenitor cells （EPCs） from human umbilical cord blood （HUCB）. Methods Mononuclearcells （MNCs） were isolated from HUCB in vitro by Ficoll density gradient centrifugation, then the cells were plated on fibronectin-coated culture dishes. Aftert 7 days, the attached cells were treated by aFGF with different concentrations （2, 5, 10 μg,/L） for 24 hours. EPCs were characterized as adherent cells with double positive to DiI-acetylated low density lipoprotein （DiIacLDL） uptake and lectin binding by direct fluorescent staining under a laser scanning confocal microscope. Flow cytom- etry was used to detect cell apoptpsis. The expressions of Bcl-2 mRNA and protein were detected respectively by reverse transcription polymerase chain reaction （RT-PCR） and Western Blot. Meanwhile, the attached cells in the group （5μg/ L aFGF group） of the most obvious effects on EPCs were cultured for 6, 12, 24, 48 h respectively, accordingly, to explore the relationship between time and effect of 5μg/L aFGF group. Results Compared with control group, aFGF can ar- gument the number of EPCs and inhibit apoptosis of EPCs （ P 〈 0. 05 ）. The increase and inhibition ratio of apoptosis reached the maximum at 24 h after administration of 5μg/L aFGF （P 〈 0. 05）. Expression of Bcl-2 mRNA and protein of EPCs in aFGF group was higher than that in control group （P 〈 0. 05）. Conclusions The results of the present study define a novel mechanism of the action of aFGF; aFGF can augment the number and inhibit apoptosis of EPCs from HUCB via up -regulating Bcl-2 expression.
出处
《中国动脉硬化杂志》
CAS
CSCD
北大核心
2013年第2期154-158,共5页
Chinese Journal of Arteriosclerosis
