μs脉冲电场诱导细胞产生的可逆电穿孔效应和不可逆电穿孔效应已有一定研究,但其所诱导的细胞凋亡效应却往往被忽略,诱导凋亡效应的脉冲电场与产生可逆/不可逆电穿孔效应脉冲电场的剂量关系更是鲜有研究。为了解这些关系,以人宫颈癌细胞系Hela为实验对象,综合运用PI示踪穿孔、CCK-8法检测活性及流式细胞术测定凋亡3种实验手段,研究了μs脉冲电场所诱导的可逆电穿孔、不可逆电穿孔及凋亡3种效应与外加脉冲场强的依赖关系。研究表明,随着外加脉冲场强的增加,细胞将依次出现电穿孔现象、凋亡现象和不可逆电穿孔现象并得出了3种效应所对应的场强取值范围(即场强窗口参数)。通过建立细胞等效电路模型以及定性分析细胞膜及线粒体膜上跨膜电压与外加电场的关系,较好地解释了实验现象及其可能机理。
The reversible electroporation(RE) and/or irreversible electroporation(IRE) phenomena induced by microsecond electric pulse fields have been focused on,whereas,little attention has been paid to the apoptosis phenomena induced by the same pulse fields.We studied the dependence of microsecond pulse field strength dose and the IRE/RE/apoptosis responses with experiments on human hela cells.Propidum Iodide(PI),CCK-8(Cell counting kit-8) assay and flow cytometry were used respectively to trace the cell electroporation,to examine survival cells,and to detect apoptosis.It is found that,with the increase of the field strength,three phenomena,such as RE,apoptosis,and IRE responses,will occur successively.Value ranges of the field strength,i.e.,filed strength windows,corresponding to IRE/RE/apoptosis are obtained.Based on the equivalent circuit model of a cell considering mitochondrion and the analysis of the transmembrane voltages of cell and of the mitochondrion,the window effect can be observed and the possible mechanisms are explained.