Edaravone protects against methylglyoxal-induced barrier damage in human brain endothelial cells

• Published in: PloS one Vol. 9; no. 7; p. e100152
• Main Authors: Tóth, Andrea E, Walter, Fruzsina R, Bocsik, Alexandra, Sántha, Petra, Veszelka, Szilvia, Nagy, Lajos, Puskás, László G, Couraud, Pierre-Olivier, Takata, Fuyuko, Dohgu, Shinya, Kataoka, Yasufumi, Deli, Mária A
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 17-07-2014; Public Library of Science (PLoS)
• Subjects: Acquired immune deficiency syndrome; AIDS; Angiogenesis; Antipyrine - analogs & derivatives; Antipyrine - pharmacology; beta Catenin - metabolism; Biochemistry, Molecular Biology; Biology and Life Sciences; Biophysics; Blood-brain barrier; Blood-Brain Barrier - drug effects; Blood-Brain Barrier - physiology; Brain; Brain damage; Brain injury; Brain research; Carbonyls; Cell Line; Cell morphology; Cell Survival - drug effects; Cerebral infarction; Claudin-5 - metabolism; Cytology; Diabetes; Electric Impedance; Endothelial cells; Endothelial Cells - physiology; Endothelium; Endothelium, Vascular - cytology; Endothelium, Vascular - drug effects; Endothelium, Vascular - pathology; Experiments; Free Radical Scavengers - pharmacology; Free radicals; Guanidines - pharmacology; Health sciences; Humans; Immunohistochemistry; Infarction; Inflammation; Integrity; Ischemia; Laboratory animals; Life Sciences; Medicine and Health Sciences; Membrane permeability; Monomolecular films; Neuroimaging; Oxidative stress; Oxidative Stress - drug effects; Permeability; Permeability - drug effects; Pharmaceuticals; Protective Agents - pharmacology; Proteins; Pyruvaldehyde; Pyruvaldehyde - pharmacology; Reactive Oxygen Species - metabolism; Research and Analysis Methods; Rodents; Stresses; Tight Junctions - drug effects; Tight Junctions - physiology; Toxicity; West Nile virus; β-Catenin; Hungary; France; Switzerland; Japan
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0100152

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.
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.
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.
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.