The generation of reactive-oxygen species associated with long-lasting pulse-induced electropermeabilisation of mammalian cells is based on a non-destructive alteration of the plasma membrane
The generation of reactive-oxygen species associated with long-lasting pulse-induced electropermeabilisation of mammalian cells is based on a non-destructive alteration of the plasma membrane
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DOI：
10.1016/s0005-2736(99)00154-6
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发表时间：
1999-11-09
影响因子：
3.4
通讯作者：
Gabriel, B
中科院分区：
文献类型：
--
作者：
Bonnafous, P;Vernhes, MC;Gabriel, B
将中国仓鼠卵巢（CHO）细胞置于适于电介导基因转移的脉冲电场中（脉冲持续时间大于1 ms）。使用化学发光探针lucigenin，我们发现当使用毫秒脉冲电渗透细胞时，存在一代活性氧（氧化跳跃）。氧化跳变产率由允许在受电影响的细胞区域内渗透的改变程度控制，但显示出对超过1ms的脉冲持续时间的饱和依赖。细胞电脉冲诱导膜组件的可逆和不可逆改变。氧化应激只有在膜渗透可逆时才会出现。不可逆的电膜破坏抑制了氧化跳跃。氧化跳变不是简单的膜电渗透反馈效应。它强烈地控制着细胞的长期存活。这必然与活性氧的细胞破坏作用有关。然而，在毫秒累积的脉冲持续时间内，大量短脉冲（微秒）的积累对细胞来说是极其致命的，而与氧化跳跃的增加没有相关性。细胞反应，如自由基的产生，存在于活细胞的电渗透过程中，并部分控制脉冲细胞的长期行为。(C) 1999 Elsevier Science B.V.版权所有
Chinese hamster ovary (CHO) cells in suspension were subjected to pulsed electric fields suitable for electrically mediated gene transfer (pulse duration longer than 1 ms). Using the chemiluminescence probe lucigenin, we showed that a generation of reactive-oxygen species (oxidative jump) was present when the cells were electropermeabilised using millisecond pulses. The oxidative jump yield was controlled by the extent of alterations allowing permeabilisation within the electrically affected cell area, but showed a saturating dependence on the pulse duration over 1 ms. Cell electropulsation induced reversible and irreversible alterations of the membrane assembly. The oxidative stress was only present when the membrane permeabilisation was reversible. Irreversible electrical membrane disruption inhibited the oxidative jump. The oxidative jump was not a simple feedback effect of membrane electropermeabilisation. It strongly controlled long-term cell survival. This had to be associated with the cell-damaging action of reactive-oxygen species. However, for millisecond-cumulated pulse duration, an accumulation of a large number of short pulses (microsecond) was extremely lethal for cells, while no correlation with an increased oxidative jump was found. Cell responses, such as the production of free radicals, were present during electropermeabilisation of living cells and controlled partially the long-term behaviour of the pulsed cell. (C) 1999 Elsevier Science B.V. All rights reserved.