Oxidative effects of nanosecond pulsed electric field exposure in cells and cell-free media.
Oxidative effects of nanosecond pulsed electric field exposure in cells and cell-free media.
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DOI：
10.1016/j.abb.2012.08.004
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发表时间：
2012-11-01
影响因子：
3.9
通讯作者：
Pakhomov, Andrei G.
中科院分区：
文献类型：
--
作者：
Pakhomova, Olga N.;Khorokhorina, Vera A.;Bowman, Angela M.;Rodaite-Riseviciene, Raminta;Saulis, Gintautas;Xiao, Shu;Pakhomov, Andrei G.
关键词：
纳秒脉冲电场（nsPEF）是一种新型的膜结构渗透和细胞内输送外源性物质的模式。我们假设，nsPEF的氧化作用可能是一个独立的主要机制，负责生物效应。通过氧化2′，7 ′-二氯二氢荧光素（H2 DCF）、二氢乙锭（DHE）或Amplex Red来评估暴露于300 ns PEF（1-13 kV/cm）的培养细胞和培养基中的ROS产生。当负载H2 DCF的细胞悬浮液经受nsPEF时，荧光2′，7 ′-二氯荧光素（DCF）的产量与脉冲数和细胞密度成比例地增加。在nsPEF敏感的Jurkat细胞中，DCF发射在暴露后随时间增加，但在nsPEF耐药的U937细胞中保持稳定。在无细胞培养基中，nsPEF促进H2 DCF转化为DCF。这种效果是不相关的加热和减少过氧化氢酶，但不是由甘露醇或超氧化物歧化酶。通过增加Amplex Red的氧化证实了nsPEF处理的培养基中H2 O2的形成。暴露于nsPEF的单个细胞内的ROS增加通过DHE的氧化而可视化。我们得出结论，nsPEF可以产生细胞外（电化学）和细胞内的ROS，包括H2 O2和可能的其他物种。因此，nsPEF的生物效应不限于电透化;同时ROS形成可能导致细胞刺激和/或氧化性细胞损伤。
Nanosecond pulsed electric field (nsPEF) is a novel modality for permeabilization of membranous structures and intracellular delivery of xenobiotics. We hypothesized that oxidative effects of nsPEF could be a separate primary mechanism responsible for bioeffects. ROS production in cultured cells and media exposed to 300-ns PEF (1–13 kV/cm) was assessed by oxidation of 2′,7′-dichlorodihydrofluoresein (H2DCF), dihidroethidium (DHE), or Amplex Red. When a suspension of H2DCF-loaded cells was subjected to nsPEF, the yield of fluorescent 2′,7′-dichlorofluorescein (DCF) increased proportionally to the pulse number and cell density. DCF emission increased with time after exposure in nsPEF-sensitive Jurkat cells, but remained stable in nsPEF-resistant U937 cells. In cell-free media, nsPEF facilitated the conversion of H2DCF into DCF. This effect was not related to heating and was reduced by catalase, but not by mannitol or superoxide dismutase. Formation of H2O2 in nsPEF-treated media was confirmed by increased oxidation of Amplex Red. ROS increase within individual cells exposed to nsPEF was visualized by oxidation of DHE. We conclude that nsPEF can generate both extracellular (electrochemical) and intracellular ROS, including H2O2 and possibly other species. Therefore, bioeffects of nsPEF are not limited to electropermeabilization; concurrent ROS formation may lead to cell stimulation and/or oxidative cell damage.
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影响因子：
13.5
作者：
Barros, LF;Stutzin, A;Hermosilla, T
通讯作者：
Hermosilla, T
影响因子：
3
作者：
Ibey, Bennett L.;Pakhomov, Andrei G.;Gregory, Betsy W.;Khorokhorina, Vera A.;Roth, Caleb C.;Rassokhin, Mikhail A.;Bernhard, Joshua A.;Wilmink, Gerald J.;Pakhomova, Olga N.
通讯作者：
Pakhomova, Olga N.
影响因子：
3.4
作者：
Korystov, Yuri N.;Shaposhnikova, Vera V.;Emel'yanov, Maksim O.
通讯作者：
Emel'yanov, Maksim O.
影响因子：
2.8
作者：
Chen, X.;Zhuang, J.;Beebe, S. J.
通讯作者：
Beebe, S. J.
影响因子：
3.4
作者：
Bonnafous, P;Vernhes, MC;Gabriel, B
通讯作者：
Gabriel, B