利用FRET技术检测不同力学条件下T细胞中ERK活性的脉冲现象  

Detection of Pulsating ERK Activity in T Cells under Biomechanical Conditions by FRET Imaging

作　　者：盛会 李亚芹 邢雨洁 邓林红[1] 欧阳明星 SHENG Hui;LI Yaqin;XING Yujie;DENG Linhong;OUYANG Mingxing(Institute of Biomedical Engineering and Health Sciences,School of Medical and Health Engineering,Changzhou University,Changzhou 213164,Jiangsu,China;School of Pharmacy&School of Biological and Food Engineering,Changzhou University,Changzhou 213164,Jiangsu,China)

机构地区：[1]常州大学医学与健康工程学院,生物医学工程与健康科学研究院,江苏常州213164 [2]常州大学药学院生物与食品工程学院,江苏常州213164

出　　处：《医用生物力学》2023年第5期924-930,共7页Journal of Medical Biomechanics

基　　金：国家自然科学基金项目(11872129,11532003);江苏省科技厅自然科学基金项目(BK20181464)。

摘　　要：目的探索Jurkat T细胞中胞外调节蛋白激酶(extracellular regulated protein kinases,ERK)活性动力学以及基质刚度对ERK活性的影响。方法利用荧光共振能量转移(fluorescence resonance energy transfer,FRET)技术实时观测Jurkat细胞中ERK活性的变化,或细胞处于I型胶原基质胶中检测其影响。结果部分Jurkat细胞中存在ERK活性脉冲现象,频率约为3次/h,FRET振幅变化约为20%。在抗体激活T细胞抗原受体(T-cell receptor,TCR)的条件下,ERK脉冲依然存在,频率和振幅无显著变化。当细胞处于I型胶原水凝胶中,随着胶基质刚度增加,脉冲频率有所下调。结论Jurkat T细胞中存在自发的ERK活性脉冲现象,初步实验显示其频率受基质刚度影响。而该信号波动的生理意义和分子机制仍有待探索。Objective To study kinetics of extracellular regulated protein kinase(ERK)activity in Jurkat T cells and the effect of matrix stiffness on ERK activity.Methods ERK activity in cells was visualized by fluorescence resonance energy transfer(FRET)biosensor,and the cells were embedded into type I collagen(COL)hydrogel to detect the biomechanical effect.Results Pulsation of ERK activity was found in a sub-group of Jurkat cells,the frequency was about three times per hour,and the average change in oscillating magnitude was about 20%.Under the condition of T-cell receptor(TCR)activation with antibodies,ERK pulse still existed,but the frequency and amplitude did not change significantly.ERK showed a decreased frequency of pulsation in COL hydrogel with an increased matrix stiffness.Conclusions ERK has autonomous pulsating activity in Jurkat T cells,and preliminary experiments show that the frequency is regulated by matrix stiffness.The physiological implication of ERK oscillation and the underlying molecular mechanism need further study.

关 键 词：T细胞 细胞外调节蛋白激酶 荧光共振能量转移 信号脉冲 基质刚度 

分 类 号：R318.01[医药卫生—生物医学工程]