Faculty of Natural Sciences, Vytautas Magnus University, Kaunas LT 44404, Lithuania.
Faculty of Natural Sciences, Vytautas Magnus University, Kaunas LT 44404, Lithuania.
Bioelectrochemistry. 2020 Oct;135:107550. doi: 10.1016/j.bioelechem.2020.107550. Epub 2020 May 13.
Control of membrane permeability to exogenous compounds by membrane electroporation can lead to cell death, which is related to permanent membrane damage, oxidation stress, leakage of intracellular molecules. In this study, we show that the predominant cell death modality after the application of high voltage electric pulses is related with inability to reseal of initial pores (first stage irreversible electroporation, FirEP). After moderately strong electric pulses, initial pores reseal, however, some cell still die later on due to electric field induced cell stress which leads to delayed cell death (late-stage irreversible electroporation, LirEP). According to our data, the period in which the majority of cells commit to either pore resealing or complete loss of barrier function depends on the intensity of electric field treatment but did not exceed 35 min. Additionally, we show that after electroporation using electric pulse parameters that induce LirEP, some cells can be rescued by supplementing medium with compounds obtained from irreversibly electroporated cells. We determined that the intracellular molecules that contribute to the increase of cell viability are larger than 30 kDa. This serves to prove that the loss of intracellular compounds plays a significant role in the decrease of cell viability after electroporation.
通过电穿孔控制细胞膜对外源化合物的通透性会导致细胞死亡，这与细胞膜的永久性损伤、氧化应激和细胞内分子的泄漏有关。在这项研究中，我们表明，应用强高压电脉冲后主要的细胞死亡方式与初始孔无法再封闭有关（第一阶段不可逆电穿孔，FirEP）。在中等强度的电脉冲后，初始孔会再封闭，但由于电场诱导的细胞应激导致一些细胞随后死亡，从而导致延迟性细胞死亡（晚期不可逆电穿孔，LirEP）。根据我们的数据，大多数细胞决定是重新封闭孔还是完全丧失屏障功能的时间取决于电场处理的强度，但不超过 35 分钟。此外，我们还表明，在用诱导 LirEP 的电脉冲参数进行电穿孔后，通过在培养基中补充从不可逆电穿孔的细胞中获得的化合物，可以拯救一些细胞。我们确定，有助于提高细胞活力的细胞内分子大于 30 kDa。这证明了在电穿孔后，细胞内化合物的丢失对细胞活力的下降起着重要作用。
