title: Edaravone protects against methylglyoxal-induced barrier damage in human brain endothelial cells.
abstract: BACKGROUND
  Elevated level of reactive carbonyl species, such as methylglyoxal, triggers carbonyl stress and activates a series of inflammatory responses leading to accelerated vascular damage. Edaravone is the active substance of a Japanese medicine, which aids neurological recovery following acute brain ischemia and subsequent cerebral infarction. Our aim was to test whether edaravone can exert a protective effect on the barrier properties of human brain endothelial cells (hCMEC/D3 cell line) treated with methylglyoxal.
  
  
  METHODS
  Cell viability was monitored in real-time by impedance-based cell electronic sensing. The barrier function of the monolayer was characterized by measurement of resistance and flux of permeability markers, and visualized by immunohistochemistry for claudin-5 and β-catenin. Cell morphology was also examined by holographic phase imaging.
  
  
  RESULTS
  Methylglyoxal exerted a time- and dose-dependent toxicity on cultured human brain endothelial cells: a concentration of 600 µM resulted in about 50% toxicity, significantly reduced the integrity and increased the permeability of the barrier. The cell morphology also changed dramatically: the area of cells decreased, their optical height significantly increased. Edaravone (3 mM) provided a complete protection against the toxic effect of methylglyoxal. Co-administration of edaravone restored cell viability, barrier integrity and functions of brain endothelial cells. Similar protection was obtained by the well-known antiglycating molecule, aminoguanidine, our reference compound.
  
  
  CONCLUSIONS
  These results indicate for the first time that edaravone is protective in carbonyl stress induced barrier damage. Our data may contribute to the development of compounds to treat brain endothelial dysfunction in carbonyl stress related diseases.
authors.affiliation: Institute of Biophysics, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary.
authors.forename: Andrea E
authors.initials: AE
authors.lastname: Tóth
authors.affiliation: Institute of Biophysics, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary.
authors.forename: Fruzsina R
authors.initials: FR
authors.lastname: Walter
authors.affiliation: Institute of Biophysics, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Bocsik
authors.affiliation: Institute of Biophysics, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary.
authors.forename: Petra
authors.initials: P
authors.lastname: Sántha
authors.affiliation: Institute of Biophysics, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary.
authors.forename: Szilvia
authors.initials: S
authors.lastname: Veszelka
authors.affiliation: Avidin Ltd., Szeged, Hungary.
authors.forename: Lajos
authors.initials: L
authors.lastname: Nagy
authors.affiliation: Avidin Ltd., Szeged, Hungary.
authors.forename: László G
authors.initials: LG
authors.lastname: Puskás
authors.affiliation: Inserm, U1016, Institut Cochin, Paris, France; CNRS, UMR8104, Paris, France; Université Paris Descartes, Sorbonne Paris Cité, Paris, France.
authors.forename: Pierre-Olivier
authors.initials: PO
authors.lastname: Couraud
authors.affiliation: Department of Pharmaceutical Care and Health Sciences, Fukuoka University, Fukuoka, Japan.
authors.forename: Fuyuko
authors.initials: F
authors.lastname: Takata
authors.affiliation: Department of Pharmaceutical Care and Health Sciences, Fukuoka University, Fukuoka, Japan.
authors.forename: Shinya
authors.initials: S
authors.lastname: Dohgu
authors.affiliation: Department of Pharmaceutical Care and Health Sciences, Fukuoka University, Fukuoka, Japan.
authors.forename: Yasufumi
authors.initials: Y
authors.lastname: Kataoka
authors.affiliation: Institute of Biophysics, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, Hungary.
authors.forename: Mária A
authors.initials: MA
authors.lastname: Deli
mesh_terms: D000983:Antipyrine / Q000031:analogs & derivatives* / Q000494:pharmacology; D001812:Blood-Brain Barrier / Q000187:drug effects / Q000502:physiology*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D062507:Claudin-5 / Q000378:metabolism; D000077553:Edaravone; D017097:Electric Impedance; D042783:Endothelial Cells / Q000502:physiology; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000473:pathology; D016166:Free Radical Scavengers / Q000494:pharmacology*; D006146:Guanidines / Q000494:pharmacology; D006801:Humans; D018384:Oxidative Stress / Q000187:drug effects; D010539:Permeability / Q000187:drug effects; D020011:Protective Agents / Q000494:pharmacology; D011765:Pyruvaldehyde / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D019108:Tight Junctions / Q000187:drug effects / Q000502:physiology*; D051176:beta Catenin / Q000378:metabolism
chemical_list: C107466:CLDN5 protein, human; D062507:Claudin-5; D016166:Free Radical Scavengers; D006146:Guanidines; D020011:Protective Agents; D017382:Reactive Oxygen Species; D051176:beta Catenin; D011765:Pyruvaldehyde; D000077553:Edaravone; C004479:pimagedine; D000983:Antipyrine