在等渗高钾培养基中孵育的HeLa细胞钾离子、氯离子和水分含量的调节变化

Regulatory changes in the K+, Cl- and water contents of HeLa cells incubated in an isosmotic high K(+)-medium.

作者信息

Ikehara T, Takahashi A, Yamaguchi H, Hosokawa K, Masuya T, Miyamoto H

机构信息

Department of Physiology, School of Medicine, University of Tokushima, Japan.

出版信息

Biochim Biophys Acta. 1991 Sep 10;1068(1):87-96. doi: 10.1016/0005-2736(91)90065-g.

DOI:10.1016/0005-2736(91)90065-g

PMID:1892858

Abstract

HeLa cells had their normal medium replaced by an isosmotic medium containing 80 mM K+, 70 mM Na+ and 100 microM ouabain. The cellular contents of K+ first increased and then decreased to the original values, that is, the cells showed a regulatory decrease (RVD) in size. The initial increase was not inhibited by various agents except by substitution of medium Cl- with gluconate. In contrast, the regulatory decrease was inhibited strongly by addition of either 1 mM quinine, 10 microM BAPTA-AM without medium Ca2+, or 0.5 mM DIDS, and partly by either 1 mM EGTA without medium Ca2+, 10 microM trifluoperazine, or substitution of medium Cl- with NO3-. Addition of DIDS to the NO3(-)-substituted medium further suppressed the K+ loss but the effect was incomplete. Intracellular Ca2+ showed a transient increase after the medium replacement. These results suggest that the initial increase in cell K+ is a phenomenon related to osmotic water movement toward Donnan equilibrium, whereas the regulatory K+ decrease is caused by K+ efflux through Ca(2+)-dependent K+ channels. The K+ decrease induced a decrease in cellular water, i.e., RVD. The K+ efflux may be more selectively associated with Cl- efflux through DIDS-sensitive channels than the efflux of other anions.

摘要

将HeLa细胞的正常培养基替换为含有80 mM K⁺、70 mM Na⁺和100 μM哇巴因的等渗培养基。细胞内K⁺含量先增加然后降至原始值，即细胞体积出现调节性减小（RVD）。除了用葡萄糖酸盐替代培养基中的Cl⁻外，各种试剂均未抑制初始增加。相反，添加1 mM奎宁、无培养基Ca²⁺时的10 μM BAPTA-AM或0.5 mM DIDS可强烈抑制调节性减小，而无培养基Ca²⁺时的1 mM EGTA、10 μM三氟拉嗪或用NO₃⁻替代培养基中的Cl⁻则部分抑制调节性减小。向用NO₃⁻替代的培养基中添加DIDS可进一步抑制K⁺流失，但效果不完全。更换培养基后，细胞内Ca²⁺出现短暂增加。这些结果表明，细胞K⁺的初始增加是一种与向唐南平衡的渗透水移动相关的现象，而调节性K⁺减少是由通过Ca²⁺依赖性K⁺通道的K⁺外流引起的。K⁺减少导致细胞内水分减少，即RVD。与其他阴离子的外流相比，K⁺外流可能更选择性地与通过DIDS敏感通道的Cl⁻外流相关。