title: Anti-thrombotic and pro-fibrinolytic effects of levosimendan in human endothelial cells in vitro.
abstract: Levosimendan is an inodilator for the treatment of acute decompensated heart failure (HF). Data from clinical studies suggest that levosimendan is particularly effective in HF due to myocardial infarction. After acute revascularization, no reflow-phenomenon is a common complication that may lead to pump failure and cardiogenic shock. Our aim was to examine whether levosimendan interferes with the pro-thrombotic phenotype of activated endothelial cells in vitro.
  
  
  
  Human heart microvascular endothelial cells (HHMEC) and human umbilical vein endothelial cells (HUVEC) were treated with interleukin-1β (IL-1β) (200U/mL) or thrombin (5U/mL) and co-treated with or without levosimendan (0.1-10μM) for 2-24h. In addition, flow experiments were performed. Effects on plasminogen activator inhibitor-1 (PAI-1) and tissue factor (TF) expression and activity were measured by rt-PCR, specific ELISA and flow cytometry.
  
  
  
  Treatment with IL-1β or thrombin significantly increased the expression of PAI-1 and TF in endothelial cells. Co-treatment with levosimendan strongly attenuated the effects of IL-1β and thrombin on PAI-1 and TF mRNA by up to 50% and 45%, in a dose- and time-dependent manner. Similar results were obtained under flow conditions. Furthermore, co-treatment with levosimendan dampened the antigen production of PAI-1 and the surface expression of TF by 35% and 45%, respectively. Additionally, levosimendan diminished both TF and PAI-1 activity.
  
  
  
  Levosimendan down-regulates the expression of the pro-thrombotic and anti-fibrinolytic biomolecules TF and PAI-1 in activated human endothelial cells. Our findings may, at least in part, explain some of the beneficial effects of levosimendan after myocardial reperfusion.
authors.affiliation: Department of Internal Medicine II, Medical University of Vienna, Austria; Ludwig Boltzmann Cluster for Cardiovascular Research, Vienna, Austria.
authors.forename: Konstantin A
authors.initials: KA
authors.lastname: Krychtiuk
authors.affiliation: Department of Internal Medicine II, Medical University of Vienna, Austria.
authors.forename: Christoph
authors.initials: C
authors.lastname: Kaun
authors.affiliation: Department of Internal Medicine II, Medical University of Vienna, Austria.
authors.forename: Philipp J
authors.initials: PJ
authors.lastname: Hohensinner
authors.affiliation: Department of Internal Medicine II, Medical University of Vienna, Austria.
authors.forename: Stefan
authors.initials: S
authors.lastname: Stojkovic
authors.affiliation: Department of Vascular Biology and Thrombosis Research, Medical University of Vienna, Austria.
authors.forename: Jacqueline
authors.initials: J
authors.lastname: Seigner
authors.affiliation: Department of Internal Medicine II, Medical University of Vienna, Austria.
authors.forename: Stefan P
authors.initials: SP
authors.lastname: Kastl
authors.affiliation: Department of Surgery, Medical University of Vienna, Austria.
authors.forename: Andreas
authors.initials: A
authors.lastname: Zuckermann
authors.affiliation: Department of Obstetrics and Gynecology, Medical University of Vienna, Austria.
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Eppel
authors.affiliation: Core Facilities, Medical University of Vienna, Austria.
authors.forename: Sabine
authors.initials: S
authors.lastname: Rauscher
authors.affiliation: Department of Vascular Biology and Thrombosis Research, Medical University of Vienna, Austria.
authors.forename: Rainer
authors.initials: R
authors.lastname: de Martin
authors.affiliation: Department of Internal Medicine II, Medical University of Vienna, Austria.
authors.forename: Gerald
authors.initials: G
authors.lastname: Maurer
authors.affiliation: 3rd Medical Department for Cardiology and Emergency Medicine, Wilhelminenhospital, Vienna, Austria.
authors.forename: Kurt
authors.initials: K
authors.lastname: Huber
authors.affiliation: Department of Internal Medicine II, Medical University of Vienna, Austria; Ludwig Boltzmann Cluster for Cardiovascular Research, Vienna, Austria; Core Facilities, Medical University of Vienna, Austria.
authors.forename: Johann
authors.initials: J
authors.lastname: Wojta
authors.affiliation: Department of Internal Medicine II, Medical University of Vienna, Austria. Electronic address: walter.speidl@meduniwien.ac.at.
authors.forename: Walter S
authors.initials: WS
authors.lastname: Speidl
mesh_terms: D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D005342:Fibrinolysis / Q000187:drug effects*; D005343:Fibrinolytic Agents / Q000494:pharmacology*; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D006835:Hydrazones / Q000494:pharmacology*; D053583:Interleukin-1beta / Q000494:pharmacology; D017395:Plasminogen Activator Inhibitor 1 / Q000378:metabolism; D011724:Pyridazines / Q000494:pharmacology*; D000077464:Simendan; D013917:Thrombin / Q000494:pharmacology; D013925:Thromboplastin / Q000378:metabolism; D013997:Time Factors
chemical_list: D005343:Fibrinolytic Agents; D006835:Hydrazones; D053583:Interleukin-1beta; D017395:Plasminogen Activator Inhibitor 1; D011724:Pyridazines; C051321:SERPINE1 protein, human; D000077464:Simendan; D013925:Thromboplastin; D013917:Thrombin
keywords: Endothelial cells; Levosimendan; PAI-1; Tissue factor