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沉默 ATP1A1 可减轻纳秒电脉冲对细胞膜的破坏。

Silencing of ATP1A1 attenuates cell membrane disruption by nanosecond electric pulses.

机构信息

Frank Reidy Research Center for Bioelectrics, Old Dominion University, Norfolk, VA, 23508, USA; Institute for Digestive System Research, Lithuanian University of Health Sciences, 44307, Kaunas, Lithuania.

Frank Reidy Research Center for Bioelectrics, Old Dominion University, Norfolk, VA, 23508, USA.

出版信息

Biochem Biophys Res Commun. 2023 Oct 15;677:93-97. doi: 10.1016/j.bbrc.2023.08.011. Epub 2023 Aug 5.

Abstract

This study explored the role of the Na/K-ATPase (NKA) in membrane permeabilization induced by nanosecond electric pulses. Using CRISPR/Cas9 and shRNA, we silenced the ATP1A1 gene, which encodes α1 NKA subunit in U937 human monocytes. Silencing reduced the rate and the cumulative uptake of YoPro-1 dye after electroporation by 300-ns, 7-10 kV/cm pulses, while ouabain, a specific NKA inhibitor, enhanced YoPro-1 entry. We conclude that the α1 subunit supports the electropermeabilized membrane state, by forming or stabilizing electropores or by hindering repair mechanisms, and this role is independent of NKA's ion pump function.

摘要

本研究探讨了钠钾-ATP 酶(NKA)在纳秒电脉冲诱导的膜通透性中的作用。我们使用 CRISPR/Cas9 和 shRNA 沉默了编码人单核细胞 U937 中 α1 NKA 亚基的 ATP1A1 基因。沉默使 300-ns、7-10 kV/cm 脉冲电穿孔后 YoPro-1 染料的摄取率和累积摄取率降低了 30%,而哇巴因是一种特异性的 NKA 抑制剂,增强了 YoPro-1 的进入。我们得出结论,α1 亚基通过形成或稳定电孔或阻碍修复机制来支持电穿孔的膜状态,而这种作用与 NKA 的离子泵功能无关。

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