title: Effects of helium on inflammatory and oxidative stress-induced endothelial cell damage.
abstract: Helium induces preconditioning in human endothelium protecting against postischemic endothelial dysfunction. Circulating endothelial microparticles are markers of endothelial dysfunction derived in response to injury. Another noble gas, xenon, protected human umbilical vein endothelial cells (HUVEC) against inflammatory stress in vitro. We hypothesised that helium protects the endothelium in vitro against inflammatory and oxidative stress. HUVEC were isolated from fresh umbilical cords and grown upon confluence. Cells were subjected to starving medium for 12h before the experiment and treated for either 3 × 5 min or 1 × 30 min with helium (5% CO2, 25% O2, 70% He) or control gas (5% CO2, 25% O2, 70% N2) in a specialised gas chamber. Subsequently, cells were stimulated with TNF-α (40 ng/ml for 24h or 10 ng/ml for 2h) or H2O2 (500 μM for 2h) or left untreated. Adhesion molecule expression was analysed using real-time quantitative polymerase chain reaction. Caspase-3 expression and viability of the cells was measured by flowcytometry. Microparticles were investigated by nanoparticle tracking analysis. Helium had no effect on adhesion molecule expression after TNF-α stimulation but in combination with oxidative stress decreased cell viability (68.9 ± 1.3% and 58 ± 1.9%) compared to control. Helium further increased TNF-α induced release of caspase-3 containing particles compared to TNF-α alone (6.4 × 10(6) ± 1.1 × 10(6) and 2.9 × 10(6) ± 0.7 × 10(6), respectively). Prolonged exposure of helium increased microparticle formation (2.4 × 10(9) ± 0.5 × 10(9)) compared to control (1.7 × 10(9) ± 0.2 × 10(9)). Summarized, helium increases inflammatory and oxidative stress-induced endothelial damage and is thus not biologically inert. A possible noxious effects on the cellular level causing alterations in microparticle formation both in number and content should be acknowledged.
authors.affiliation: Academic Medical Centre (AMC), Department of Anesthesiology, Laboratory of Experimental Intensive Care and Anesthesiology (L.E.I.C.A.), Academic Medical Centre (AMC), University of Amsterdam, Meibergdreef 9, 1100 DD Amsterdam, The Netherlands.
authors.forename: Kirsten F
authors.initials: KF
authors.lastname: Smit
authors.affiliation: Academic Medical Centre (AMC), Department of Anesthesiology, Laboratory of Experimental Intensive Care and Anesthesiology (L.E.I.C.A.), Academic Medical Centre (AMC), University of Amsterdam, Meibergdreef 9, 1100 DD Amsterdam, The Netherlands.
authors.forename: Raphaela P
authors.initials: RP
authors.lastname: Kerindongo
authors.affiliation: Department of Experimental Clinical Chemistry, Academic Medical Centre (AMC), Meibergdreef 9, 1100 DD Amsterdam, The Netherlands.
authors.forename: Anita
authors.initials: A
authors.lastname: Böing
authors.affiliation: Department of Experimental Clinical Chemistry, Academic Medical Centre (AMC), Meibergdreef 9, 1100 DD Amsterdam, The Netherlands.
authors.forename: Rienk
authors.initials: R
authors.lastname: Nieuwland
authors.affiliation: Academic Medical Centre (AMC), Department of Anesthesiology, Laboratory of Experimental Intensive Care and Anesthesiology (L.E.I.C.A.), Academic Medical Centre (AMC), University of Amsterdam, Meibergdreef 9, 1100 DD Amsterdam, The Netherlands.
authors.forename: Markus W
authors.initials: MW
authors.lastname: Hollmann
authors.affiliation: Academic Medical Centre (AMC), Department of Anesthesiology, Laboratory of Experimental Intensive Care and Anesthesiology (L.E.I.C.A.), Academic Medical Centre (AMC), University of Amsterdam, Meibergdreef 9, 1100 DD Amsterdam, The Netherlands.
authors.forename: Benedikt
authors.initials: B
authors.lastname: Preckel
authors.affiliation: Academic Medical Centre (AMC), Department of Anesthesiology, Laboratory of Experimental Intensive Care and Anesthesiology (L.E.I.C.A.), Academic Medical Centre (AMC), University of Amsterdam, Meibergdreef 9, 1100 DD Amsterdam, The Netherlands. Electronic address: N.C.Hauck@amc.uva.nl.
authors.forename: Nina C
authors.initials: NC
authors.lastname: Weber
mesh_terms: D017209:Apoptosis; D002317:Cardiovascular Agents / Q000494:pharmacology*; D053148:Caspase 3 / Q000378:metabolism; D055252:Cell-Derived Microparticles / Q000378:metabolism; D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical; D019040:E-Selectin / Q000378:metabolism; D006371:Helium / Q000494:pharmacology*; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism
chemical_list: D002317:Cardiovascular Agents; D019040:E-Selectin; D019010:Vascular Cell Adhesion Molecule-1; D018799:Intercellular Adhesion Molecule-1; D006371:Helium; C505407:CASP3 protein, human; D053148:Caspase 3
keywords: Human umbilical vein endothelial cells; Inflammatory stress; Microparticles; Noble gases; Preconditioning