高功率脉冲微波和电磁脉冲辐照对心肌细胞膜电穿孔效应及机理的研究

The Electroporation Effects of High Power Pulse Microwave and Electromagnetic Pulse Irradiation on the Membranes of Cardiomyocyte Cells and the Mechanism Therein Involved

电磁辐射可造成人体多器官系统的损伤。对其损伤的病理机制至今尚未完全阐明。心脏是电磁辐射损伤的敏感器官之一。本实验拟通过研究HPPM和EMP对心肌细胞的影响,探讨电磁辐射致伤的关键机制,解释脉冲电磁场致心肌细胞损伤的发展规律。本实验采用HPPM和EMP辐照原代培养心肌细胞的方法,用原子力显微镜、激光扫描共聚焦显微镜、流式细胞仪和全自动生化检测仪等多种实验手段,检测辐照对细胞膜形态、结构和功能的影响。结果发现HPPM和EMP辐照后,心肌细胞搏动减慢,形态异常,活力下降,凋亡和坏死率明显增加。细胞膜出现数量不等、大小不一的电穿孔,细胞膜通透性丧失,结构破坏,细胞培养液中Na+、K+、Ca2+、Cl-、Mg2+、Ca2+和无机磷离子浓度显著升高(P<0.01),细胞内Ca2+浓度显著降低(P<0.01)。结果提示心肌细胞是电磁辐射损伤的敏感细胞,脉冲电磁场可致心肌细胞膜电穿孔,细胞形态、结构和功能均受到严重损伤。电穿孔效应是解释电磁辐射非热效应的关键机制之一。电穿孔导致的细胞膜通透性和膜结构改变存在一定发展规律。

Though there is ongoing public concern on potential hazards and risk of electromagnetic radiation, the bioeffects mechanism of electromagnetic fields remains obscure. Heart is one of the organs susceptive to electromagnetic fields （EMF）. This study was designed to assess the influence of high power pulse microwave and electromagnetic pulse irradiation on cardiomyocytes, to explore the critical mechanism of electromagnetic fields, and to explain the regular course of injury caused by exposure to pulse EMF. Cultured cardiomyocytes were irradiated by high power pulse microwave and electromagnetic pulse first, then a series of apparatus including atom force microscope, laser scanning confocal microscope and flow cytometer were used to examine the changes of cell membrane conformation, structure and function. After irradiation, the cardiomyocytes pulsated slower or stop,the cells conformation was abnormal, the cells viability declined, and the percentage of apoptosis and necrosis increased significantly （P〈 0. 01 ）. The cell membrane had pores unequal in size, and lost its penetration character. The concentration of Na^＋, K^＋, Ca^2＋, C^l-, Mg^2＋, Ca^2＋ and P^3＋ in cell culture medium increased significantly （P〈0.01）. and the concentration of Ca^2＋ in cells （[Ca^2＋]i） decreased significantly （P〈0.01）. The results indicated that cardiomyocytes are susceptible to non-ionizing radiation. Pulse electromagnetic field can induce cardiomyocytes electroporation, and can do great damage to cells conformation, structure and function. Electroporation is one of the most critical mechanisms to explain the athermal effects of electromagnetic radiation.