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从分子和细胞水平探讨机体对外界刺激的反应机理,对从微观水平研究活细胞内分子活动的过程、规律及特点以及开展细胞分子生物学研究具有重要意义.本研究针对动态监测和研究活细胞内分子事件的需求,以表面等离子体共振(surfaceplasmonresonance,SPR)技术为核心,设计研制了能够用于细胞培养和动态细胞学效应监测的SPR细胞传感器装置.利用该装置,实时研究了佛波酯作用于C6细胞所诱发的特征性SPR信号.该信号同PMA作用于无细胞的空白传感器明显不同,呈现规律性变化,具有PMA剂量依赖性和饱和性特点,且能够被PKC抑制剂减弱甚至完全抑制.这说明该信号源于PKC活化等细胞内反应所导致的生物分子在细胞膜部位的重新分布.SPR细胞传感器技术为动态、无创性研究活细胞内分子事件发生的时空特点及规律提供了新的量化分析方法与思路.该技术还可用于空间细胞学效应研究、配体垂钓、药物筛选及生物战剂监测等基础研究和应用研究.
It is of great importance to study the reaction mechanism of organism to external stimuli at the molecular and cellular level and to study the process, regularity and characteristics of molecular activity in living cells at the micro level and to carry out molecular molecular biology research.In this study, the dynamic monitoring and study of living cells In order to meet the needs of intramolecular events, the SPR cell sensor device designed for cell culture and dynamic cytology monitoring was designed and developed based on the surface plasmon resonance (SPR) technique. With this device, Acts on the characteristic SPR signal induced by C6 cells, which is obviously different from PMA on cell-free blank sensor, showing regular changes, having the dose-dependent and saturation characteristics of PMA, and can be weakened even completely by PKC inhibitors Inhibition, indicating that this signal is derived from the redistribution of biomolecules at the cell membrane site caused by intracellular reactions such as PKC activation.Spring Cell Sensor technology provides a new, dynamic and noninvasive way to study the temporal and spatial characteristics and patterns of molecular events in living cells Quantitative analysis methods and ideas.The technology can also be used for space cytology We should study, ligand fishing, drug screening and basic research biological warfare agents such as monitoring and applied research.