摘要
目的探讨microRNA-214-3p(miR-214-3p)在周期性张应变诱导内皮祖细胞(endothelial progenitor cells, EPCs)分化和增殖中的作用。方法采用FX-5000T细胞周期性张应变加载装置对EPCs施加生理水平的周期性张应变(5%幅度、1.25 Hz频率),加载时间24 h。应用miRNAs芯片筛选周期性张应变调控下差异表达的miRNAs,并挑选miR-214-3p进行深入研究。实时荧光定量PCR方法检测EPCs内平滑肌细胞(vascular smooth muscle cells, VSMCs)相标志分子的表达,BrdU结合酶联免疫吸附ELISA法检测EPCs增殖功能。之后,使用miR-214-3p抑制剂抑制miR-214-3p的表达,检测EPC内VSMC相标志分子表达及EPCs增殖。结果周期性张应变显著抑制miR-214-3p表达,并抑制EPCs向VSMC相分化,同时显著促进EPCs增殖。在静态条件下,使用miR-214-3p抑制剂干扰miR-214-3p的表达,miR-214-3p水平下降同样会抑制EPCs向VSMC相分化,并且诱导EPCs增殖能力显著上升。结论生理水平的周期性张应变能够抑制EPCs内miR-214-3p表达,从而抑制EPCs向VSMC相分化,并且促进EPCs增殖。研究结果为血管损伤的治疗提供新的治疗靶点。
Objective To investigate the role of microRNA-214-3p(miR-214-3p)in differentiation and proliferation of endothelial progenitor cells(EPCs)induced by cyclic stretch.Methods EPCs were exposed to cyclic stretch at physiological level(with the magnitude of 5%,at a constant frequency of 1.25 Hz)for 24 h by FX-5000T Strain Unit.miRNAs array was performed to identify the expression profiling of miRNAs.Real-time PCR was used to examine the expression levels of miRs.The expression of vascular smooth muscle cells(VSMCs)markers in EPCs was detected by real-time PCR.EPC proliferation was detected by BrdU ELISA assay.After EPCs were transfected with miR-214-3p inhibitor(IN)to knockdown expression of miR-214-3p,the level of VSMC markers expression and EPC proliferation was detected.Results Cyclic stretch significantly decreased miR-214-3p expression,depressed EPC differentiation toward VSMCs,and increased EPCs proliferation.Similarly,transfection with the miR-214-3p inhibitor led to the decreased expression of VSMC markers under static station.Meanwhile,miR-214-3p down-regulation promoted EPC proliferation significantly.Conclusions Physiological cyclic stretch could down-regulate the expression of miR-214-3p in EPCs,depress EPC differentiation towards VSMC and promote EPC proliferation eventually.Therefore,the research findings provide a potential therapeutic strategy for treating vessel injuries.
作者
李娜
汪文斌
阎靖
齐颖新
韩悦
LI Na;WANG Wenbin;YAN Jing;QI Yingxin;HAN Yue(Institute of Mechanobiology and Medical Engineering,School of Life Sciences and Biotechnology,Shanghai Jiao Tong University,Shanghai 200240,China)
出处
《医用生物力学》
EI
CAS
CSCD
北大核心
2019年第1期69-76,共8页
Journal of Medical Biomechanics
基金
国家自然科学基金项目(11572198
11625209
11572199)
关键词
周期性张应变
内皮祖细胞
细胞增殖
细胞分化
cyclic stretch
endothelial progenitor cells
cell proliferation
cell differentiation
