电工技术学报  2023, Vol. 38 Issue (14): 3779-3788    DOI: 10.19595/j.cnki.1000-6753.tces.220786
电工理论 |
脉冲电场对细胞膜电穿孔面积的影响研究
吴晓1,2, 米彦1, 郑伟1, 马驰1, 张梦男1
1.输配电装备及系统安全与新技术国家重点实验室(重庆大学) 重庆 400030;
2.国网河南省电力公司营销服务中心(计量中心) 郑州 450000
Effect of Pulsed Electric Field on Electroporation Area of Cell Membrane
Wu Xiao1,2, Mi Yan1, Zheng Wei1, Ma Chi1, Zhang Mengnan1
1. State Key Laboratory of Power Transmission Equipment & System Security and New Technology Chongqing University Chongqing 400030 China;
2. Marketing Service Center of State Grid Henan Electric Power Company (Metrology Center) Zhengzhou 450000 China
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摘要 目前在实验中通常采用间接的表征参量以衡量细胞电穿孔效应,为了能够在实验中找到一种直接的表征参量,该文提出了电穿孔面积这一新的表征参量及其测量方法。基于分子传输动力学,在脉冲电场下开展了荧光分子染色实验,然后通过对细胞荧光强度曲线进行数据处理,计算出脉冲期间形成的电穿孔面积,并研究了不同脉冲电场参数下的细胞膜电穿孔面积变化规律。结果表明,随着脉冲电场参数增强,细胞荧光强度增大,电穿孔效应增强,计算出的电穿孔面积数值与已有结论吻合,证实了该实验方法的可行性。此外,电穿孔面积随着电场强度、脉冲宽度和脉冲电冲击的增大而线性增大,为脉冲电场临床治疗提供了指导。
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吴晓
米彦
郑伟
马驰
张梦男
关键词 脉冲电场电穿孔效应电穿孔面积分子传输荧光染色实验    
Abstract:In the field of electrobiology, simulation and experimental methods can be used to study the cell electroporation effect under pulsed electric field. In simulation, parameters such as transmembrane voltage, current density, pore density, pore radius and pore area are usually used to characterize the electroporation effect. In the experiment, the killing effect or the overall situation of the electric field on the cells is usually characterized macroscopically, which is not equivalent to the specific parameters such as the number and size of the pores formed, and it is not easy to establish a close relationship between simulation and experiment. Therefore, the electroporation area of cell membrane, a new characterization parameter to directly measure the electroporation effect, and its experimental measurement method are proposed in this paper.
Firstly, the measurement principle of cell electroporation area was analyzed based on the molecular transport dynamics, and it was found that the electroporation area could be measured and calculated by cell fluorescence experiment. Then, the fluorescent molecular staining experiment was carried out based on the above principle, the fluorescence intensity changes of cells under the electric field were captured in real time under the fluorescence microscope. Before carrying out cell electroporation experiments, the cell fluorescence intensity needs to be corrected to obtain the relationship between the cell fluorescence intensity and the intracellular concentration of propidium iodide ions. Next, the maximum cell fluorescence intensity and the pore recovery time constant were extracted from the fluorescence intensity curve under the action of the electric field, from which the electroporation area of cell membrane formed during the pulse could be calculated. Finally, experiments were carried out under different pulsed electric field parameters (field strength and pulse width) to study the relationship and law between cell electroporation area and pulsed electric field parameters, and the obtained laws were analyzed and explained theoretically.
Under the pulsed electric field parameters selected in this paper, the electroporation area calculated by the cell fluorescent staining experiment is in the order of 10-15 m2, which is quantitatively close to the value calculated in the literature. Under different pulse electric field parameters, with the increase of pulse electric field parameters (field strength and pulse width), the fluorescence intensity of cells increases, the fluorescence intensity curve moves up, and the electroporation effect is enhanced. Moreover, the calculated electroporation area increases with the increase of pulse field strength, pulse width and pulse electric shock, and all have a linear relationship. The obtained law is analyzed in combination with theory, and the laws obtained in this paper are qualitatively consistent with those in the existing literature. By comparing the electroporation area between this paper and the existing conclusions quantitatively and qualitatively, the accuracy of the method and results of calculating the cell electroporation area by experimental measurements can be evaluated. According to these laws, it can provide guidance for the pulsed electric field treatment. For example, in the electrochemotherapy, the pulsed electric field parameters (field strength and pulse width) are increased linearly, the electroporation area is increased, and more drugs enter the cells, and the treatment is more effective.
Key wordsPulsed electric field    electroporation effect    electroporation area    molecular transport    fluorescent staining experiment   
收稿日期: 2022-05-10     
PACS: TM836  
  Q64  
基金资助:国家自然科学基金项目(52077022, 51477022)和“111”引智项目(BP0820005)资助
通讯作者: 米 彦 男,1978年生,教授,博士生导师,研究方向为高电压新技术。E-mail:miyan@cqu.edu.cn   
作者简介: 吴 晓 女,1996年生,硕士研究生,研究方向为高电压新技术。E-mail:20191102061t@cqu.edu.cn
引用本文:   
吴晓, 米彦, 郑伟, 马驰, 张梦男. 脉冲电场对细胞膜电穿孔面积的影响研究[J]. 电工技术学报, 2023, 38(14): 3779-3788. Wu Xiao, Mi Yan, Zheng Wei, Ma Chi, Zhang Mengnan. Effect of Pulsed Electric Field on Electroporation Area of Cell Membrane. Transactions of China Electrotechnical Society, 2023, 38(14): 3779-3788.
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