title: Time- and dose dependent actions of cardiotonic steroids on transcriptome and intracellular content of Na+and K+: a comparative analysis.
abstract: Recent studies demonstrated that in addition to Na+,K+-ATPase inhibition cardiotonic steroids (CTSs) affect diverse intracellular signaling pathways. This study examines the relative impact of [Na+]i/[K+]i-mediated and -independent signaling in transcriptomic changes triggered by the endogenous CTSs ouabain and marinobufagenin (MBG) in human umbilical vein endothelial cells (HUVEC). We noted that prolongation of incubation increased the apparent affinity for ouabain estimated by the loss of [K+]iand gain of [Na+]i. Six hour exposure of HUVEC to 100 and 3,000 nM ouabain resulted in elevation of the [Na+]i/[K+]iratio by ~15 and 80-fold and differential expression of 258 and 2185 transcripts, respectively. Neither [Na+]i/[K+]iratio nor transcriptome were affected by 6-h incubation with 30 nM ouabain. The 96-h incubation with 3 nM ouabain or 30 nM MBG elevated the [Na+]i/[K+]iratio by ~14 and 3-fold and led to differential expression of 880 and 484 transcripts, respectively. These parameters were not changed after 96-h incubation with 1 nM ouabain or 10 nM MBG. Thus, our results demonstrate that elevation of the [Na+]i/[K+]iratio is an obligatory step for transcriptomic changes evoked by CTS in HUVEC. The molecular origin of upstream [Na+]i/[K+]isensors involved in transcription regulation should be identified in forthcoming studies.
authors.affiliation: Faculty of Biology, M. V. Lomonosov Moscow State University, Moscow, 119234, Russia.
authors.forename: Elizaveta A
authors.initials: EA
authors.lastname: Klimanova
authors.affiliation: Faculty of Biology, M. V. Lomonosov Moscow State University, Moscow, 119234, Russia.
authors.forename: Artem M
authors.initials: AM
authors.lastname: Tverskoi
authors.affiliation: Faculty of Biology, M. V. Lomonosov Moscow State University, Moscow, 119234, Russia.
authors.forename: Svetlana V
authors.initials: SV
authors.lastname: Koltsova
authors.affiliation: Faculty of Biology, M. V. Lomonosov Moscow State University, Moscow, 119234, Russia.
authors.forename: Svetlana V
authors.initials: SV
authors.lastname: Sidorenko
authors.affiliation: Faculty of Biology, M. V. Lomonosov Moscow State University, Moscow, 119234, Russia.
authors.forename: Olga D
authors.initials: OD
authors.lastname: Lopina
authors.affiliation: Research Centre, University of Montreal Hospital (CRCHUM), Montreal, H2X 0A9, Canada.
authors.forename: Johanne
authors.initials: J
authors.lastname: Tremblay
authors.affiliation: Research Centre, University of Montreal Hospital (CRCHUM), Montreal, H2X 0A9, Canada.
authors.forename: Pavel
authors.initials: P
authors.lastname: Hamet
authors.affiliation: National Research Tomsk State University, Tomsk, 634050, Russia.
authors.forename: Leonid V
authors.initials: LV
authors.lastname: Kapilevich
authors.affiliation: Faculty of Biology, M. V. Lomonosov Moscow State University, Moscow, 119234, Russia.|National Research Tomsk State University, Tomsk, 634050, Russia.
authors.forename: Sergei N
authors.initials: SN
authors.lastname: Orlov
mesh_terms: D002018:Bufanolides / Q000494:pharmacology; D002301:Cardiac Glycosides / Q000494:pharmacology*; D002316:Cardiotonic Agents / Q000494:pharmacology*; D002460:Cell Line; D005786:Gene Expression Regulation / Q000187:drug effects; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D007477:Ions / Q000378:metabolism*; D010042:Ouabain / Q000494:pharmacology; D011188:Potassium / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D012964:Sodium / Q000378:metabolism*; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects; D059467:Transcriptome / Q000187:drug effects*
chemical_list: D002018:Bufanolides; D002301:Cardiac Glycosides; D002316:Cardiotonic Agents; D007477:Ions; C093896:marinobufagenin; D010042:Ouabain; D012964:Sodium; D000254:Sodium-Potassium-Exchanging ATPase; D011188:Potassium