TY - JOUR
T1 - Elevation of intracellular Na+ contributes to expression of early response genes triggered by endothelial cell shrinkage
AU - Shiyan, Alexandra A.
AU - Sidorenko, Svetlana V.
AU - Fedorov, Dmitry
AU - Klimanova, Elizaveta A.
AU - Smolyaninova, Larisa V.
AU - Kapilevich, Leonid V.
AU - Grygorczyk, Ryszard
AU - Orlov, Sergei N.
N1 - Funding Information:
This work was supported by grants from the Russian Foundation for Basic Research ( 匃猃稁爃瘁爃爃爃砃甁 to SNO) and Natural Sciences and Engineering Research Council of Canada (RGPIN 瘃甃眃眃猃礁球爃猃甁 to RG).
Publisher Copyright:
© 2019 The Author(s). Published by Cell Physiol Biochem Press GmbH&Co. KG
Copyright:
Copyright 2019 Elsevier B.V., All rights reserved.
PY - 2019
Y1 - 2019
N2 - Background/Aims: Prolonged hyperosmotic shrinkage evokes expression of osmoprotective genes via nuclear factor NFAT5-mediated pathway and activates Na+ influx via hypertonicity-induced cation channels (HICC). In human umbilical vein endothelial cells (HUVEC) elevation of intracellular sodium concentration ([Na+]i) triggers transcription of dozens of early response genes (ERG). This study examined the role of monovalent cations in the expression of Na+ isensitive ERGs in iso- and hyperosmotically shrunken HUVEC. Methods: Cell volume was measured by 3D reconstruction of cell shape and as 14C-urea available space. Intracellular Na+ and K+ content was measured by flame atomic absorption spectrometry. ERG transcription was estimated by RT-PCR. Results: Elevation of medium osmolality by 150 mM mannitol or cell transfer from hypo- to isosmotic medium decreased cell volume by 40-50%. Hyperosmotic medium increased [Na+]i by 2-fold whereas isosmotic shrinkage had no impact on this parameter. Hyperosmotic but not isosmotic shrinkage increased up-to 5-fold the content of EGR1, FOS, ATF3, ZFP36 and JUN mRNAs. Expression of these ERGs triggered by hyperosmotic shrinkage and Na+,K+-ATPase inhibition by 0.1 µM ouabain exhibited positive correlation (R2=0.9383, p=0.0005). Isosmotic substitution of NaCl by N-methyl-D-glucamine abolished an increment of [Na+]i and ERG expression triggered by mannitol addition. Conclusion: Augmented expression of ERGs in hyperosmotically shrunken HUVEC is mediated by elevation of [Na+]i
AB - Background/Aims: Prolonged hyperosmotic shrinkage evokes expression of osmoprotective genes via nuclear factor NFAT5-mediated pathway and activates Na+ influx via hypertonicity-induced cation channels (HICC). In human umbilical vein endothelial cells (HUVEC) elevation of intracellular sodium concentration ([Na+]i) triggers transcription of dozens of early response genes (ERG). This study examined the role of monovalent cations in the expression of Na+ isensitive ERGs in iso- and hyperosmotically shrunken HUVEC. Methods: Cell volume was measured by 3D reconstruction of cell shape and as 14C-urea available space. Intracellular Na+ and K+ content was measured by flame atomic absorption spectrometry. ERG transcription was estimated by RT-PCR. Results: Elevation of medium osmolality by 150 mM mannitol or cell transfer from hypo- to isosmotic medium decreased cell volume by 40-50%. Hyperosmotic medium increased [Na+]i by 2-fold whereas isosmotic shrinkage had no impact on this parameter. Hyperosmotic but not isosmotic shrinkage increased up-to 5-fold the content of EGR1, FOS, ATF3, ZFP36 and JUN mRNAs. Expression of these ERGs triggered by hyperosmotic shrinkage and Na+,K+-ATPase inhibition by 0.1 µM ouabain exhibited positive correlation (R2=0.9383, p=0.0005). Isosmotic substitution of NaCl by N-methyl-D-glucamine abolished an increment of [Na+]i and ERG expression triggered by mannitol addition. Conclusion: Augmented expression of ERGs in hyperosmotically shrunken HUVEC is mediated by elevation of [Na+]i
KW - Cell shrinkage
KW - Early response genes
KW - Endothelium
KW - Intracellular Na
KW - Ouabain
KW - Transcription
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U2 - 10.33594/000000162
DO - 10.33594/000000162
M3 - Article
C2 - 31556253
AN - SCOPUS:85072662789
VL - 53
SP - 638
EP - 647
JO - Cellular Physiology and Biochemistry
JF - Cellular Physiology and Biochemistry
SN - 1015-8987
IS - 4
ER -