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,随着胶的刚度增加,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 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 underlying molecular mechanism need further study.

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)