title: The phosphodiesterase III inhibitor cilostazol ameliorates ethanolinduced endothelial dysfunction.
abstract: Intracranial hemorrhage (ICH) remains a devastating disease, and heavy alcohol consumption is an underlying risk factor. The aim of this study was to study the mechanism of ethanol-induced endothelial cell damage and to evaluate the protective effect of cilostazol against ethanol-induced damage. We first evaluated transendothelial electrical resistance (TEER) and cell viability of human brain microvascular endothelial cells at the ethanol concentration shown to cause mild-to-moderate intoxication in the clinic. We also assessed the permeability of fluorescein isothiocyanate (FITC)- dextran and the change in tight junction proteins. Furthermore, we studied the potential of cilostazol to protect endothelial cells from ethanol-induced dysfunction. Concentration- and time-dependent effects of ethanol on cell viability and TEER showed that TEER was reduced at each concentration of ethanol tested during exposures of >2 h, but cell viability was not changed. Permeability of FITC-dextran was enhanced, and both tight junction and adherens junction proteins were reduced by 3-h ethanol treatment. The permeability of FITC-dextran was ameliorated by administration of cilostazol in a concentration-dependent manner. The protective effect of cilostazol was obstructed by administration of a protein kinase A inhibitor. Using gelatin zymography, we found that the protective effect of cilostazol was by reducing matrix metalloproteinase 9 (MMP-9) activation, but it had no effect on reactive oxygen spices (ROS). Our results indicate that cilostazol protected endothelial cells against ethanol-induced endothelial dysfunction by inhibiting ROS-mediated activation of MMP-9.
authors.forename: Toshinori
authors.initials: T
authors.lastname: Takagi
authors.forename: Keisuke
authors.initials: K
authors.lastname: Mishiro
authors.forename: Masamitsu
authors.initials: M
authors.lastname: Shimazawa
authors.forename: Shinichi
authors.initials: S
authors.lastname: Yoshimura
authors.forename: Toru
authors.initials: T
authors.lastname: Iwama
authors.affiliation: Department of Biofunctional Evaluation, Molecular Pharmacology, Gifu Pharmaceutical University, 1- 25-4 Daigaku-nishi, Gifu 501-1196, Japan. hidehara@gifu-pu.ac.jp.
authors.forename: Hideaki
authors.initials: H
authors.lastname: Hara
mesh_terms: D022005:Adherens Junctions / Q000187:drug effects; D001808:Blood Vessels / Q000166:cytology; D001921:Brain / Q000166:cytology; D002463:Cell Membrane Permeability; D002470:Cell Survival; D002478:Cells, Cultured; D002492:Central Nervous System Depressants / Q000494:pharmacology*; D000077407:Cilostazol; D003911:Dextrans / Q000493:pharmacokinetics; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects*; D000431:Ethanol / Q000494:pharmacology*; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000493:pharmacokinetics; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D058987:Phosphodiesterase 3 Inhibitors / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D013777:Tetrazoles / Q000494:pharmacology*; D013997:Time Factors; D062826:Zonula Occludens-1 Protein / Q000378:metabolism
chemical_list: D002492:Central Nervous System Depressants; D003911:Dextrans; D058987:Phosphodiesterase 3 Inhibitors; D017382:Reactive Oxygen Species; C569042:TJP1 protein, human; D013777:Tetrazoles; D062826:Zonula Occludens-1 Protein; C015219:fluorescein isothiocyanate dextran; D000431:Ethanol; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D016650:Fluorescein-5-isothiocyanate; D000077407:Cilostazol