Leibniz Institute for Plasma Science and Technology, Greifswald, Germany.
Leibniz Institute for Plasma Science and Technology, Greifswald, Germany; Research Centre for Toxic Compounds in the Environment (RECETOX), Faculty of Science, Masaryk University, Brno, Czech Republic.
Bioelectrochemistry. 2016 Dec;112:33-46. doi: 10.1016/j.bioelechem.2016.07.003. Epub 2016 Jul 10.
Gap junctional intercellular communication (GJIC) is an important mechanism that is involved and affected in many diseases and injuries. So far, the effect of nanosecond pulsed electric fields (nsPEFs) on the communication between cells was not investigated. An in vitro approach is presented with rat liver epithelial WB-F344 cells grown and exposed in a monolayer. In order to observe sub-lethal effects, cells were exposed to pulsed electric fields with a duration of 100ns and amplitudes between 10 and 20kV/cm. GJIC strongly decreased within 15min after treatment but recovered within 24h. Gene expression of Cx43 was significantly decreased and associated with a reduced total amount of Cx43 protein. In addition, MAP kinases p38 and Erk1/2, involved in Cx43 phosphorylation, were activated and Cx43 became hyperphosphorylated. Immunofluorescent staining of Cx43 displayed the disassembly of gap junctions. Further, a reorganization of the actin cytoskeleton was observed whereas tight junction protein ZO-1 was not significantly affected. All effects were field- and time-dependent and most pronounced within 30 to 60min after treatment. A better understanding of a possible manipulation of GJIC by nsPEFs might eventually offer a possibility to develop and improve treatments.
缝隙连接细胞间通讯（GJIC）是一种重要的机制，涉及并影响许多疾病和损伤。到目前为止，还没有研究纳秒级脉冲电场（nsPEFs）对细胞间通讯的影响。本研究采用体外方法，使用单层培养的大鼠肝上皮 WB-F344 细胞进行实验。为了观察亚致死效应，细胞暴露于持续时间为 100ns、幅度在 10 到 20kV/cm 的脉冲电场中。处理后 15 分钟内 GJIC 明显下降，但在 24 小时内恢复。Cx43 的基因表达显著降低，与 Cx43 蛋白总量减少有关。此外，参与 Cx43 磷酸化的 MAP 激酶 p38 和 Erk1/2 被激活，Cx43 发生过度磷酸化。Cx43 的免疫荧光染色显示缝隙连接解体。此外，还观察到肌动蛋白细胞骨架的重排，而紧密连接蛋白 ZO-1 没有明显受到影响。所有效应均具有场依赖性和时间依赖性，在处理后 30 至 60 分钟内最为明显。更好地了解 nsPEFs 对 GJIC 的可能调控作用，最终可能为开发和改进治疗方法提供可能。
