临床口腔医学杂志2025，Vol.41Issue(4)：195-201,7.DOI:10.3969/j.issn.1003-1634.2025.04.002
低浓度ROS对尖端细胞分化的影响及其机制探究
The effect of low concentration ROS on the differentiation of tip cells and its mechanism
摘要
Abstract
Objective:To investigate the effects of low-concentration reactive oxygen species(ROS)on tip cell differ-entiation and elucidate the underlying mechanisms.Methods:The optimal H2O2 concentration mimicking the oxidative stress microenvironment was screened using the CCK-8 assay to evaluate its impact on human umbilical vein endothelial cells(hUVECs).Tip cell differentiation was systematically assessed through CD34 immunofluorescence staining,quantitative re-verse transcription polymerase chain reaction(qRT-PCR)analysis of tip cell markers,flow cytometry to determine CD34+cell proportions,and a spheroid sprouting assay.Glycolytic activity in hUVECs under low-concentration H2O2 was analyzed by measuring glycolytic marker gene expression(including PFKFB3,HK1,PKLR,and PFKM)and lactate production.The glyco-lytic inhibitor oxamate was employed to validate its suppressive effects on glycolysis.Finally,oxamate-mediated glycolysis in-hibition was utilized to reverse-verify the role of H2O2 in tip cell differentiation.Results:The CCK-8 assay identified 0.5 mmol/L H2O2 as the optimal concentration for simulating oxidative stress.The H2O2-treated group exhibited significantly higher intracellular ROS fluorescence intensity compared to the control.Additionally,the H2O2 group demonstrated elevated CD34 fluorescence intensity,increased CD34+cell proportion,enhanced spheroid sprouting number and invasion distance,and upregulated expression of tip cell markers.H2O2 treatment also markedly increased intracellular lactate levels and the expres-sion of key glycolytic enzymes.In contrast,the oxamate+H2O2 co-treatment group showed significantly reduced lactate content and glycolytic enzyme expression compared to the H2O2 group.Furthermore,oxamate+H2O2 co-treatment resulted in dimin-ished CD34 fluorescence intensity,CD34+cell proportion,spheroid sprouting capacity,and tip cell marker expression relative to H2O2 treatment alone.Conclusion:Low-concentration ROS promotes tip cell differentiation by enhancing glycolytic activity.关键词
尖端细胞/血管再生/活性氧/内皮细胞Key words
Tip cell/Vascular regeneration/Reactive oxygen species/Endothelial cell分类
生物工程引用本文复制引用
刘方勇,张时菡,郭亚茹,邓旭亮..低浓度ROS对尖端细胞分化的影响及其机制探究[J].临床口腔医学杂志,2025,41(4):195-201,7.基金项目
国家自然科学基金青年项目(82201123) （82201123）
