title: Neuroprotective effects of cilostazol are mediated by multiple mechanisms in a mouse model of permanent focal ischemia.
abstract: The phosphodiesterase (PDE) 3 inhibitor cilostazol, used as an anti-platelet drug, reportedly can also ameliorate ischemic brain injury. Here, we investigated the effects of cilostazol in a permanent focal ischemia mice model. Male Balb/c mice were subjected to permanent middle cerebral artery occlusion. Mice were then treated with either cilostazol (10 or 20mg/kg) or vehicle administered at 30min and 24h post-ischemia, and infarct volumes were assessed at 48h post-ischemia. Mice treated with 20mg/kg of cilostazol or vehicle were sacrificed at 6h or 24h post-ischemia and immunohistochemistry was used for brain sections. Treatment with 20mg/kg of cilostazol significantly reduced infarct volumes to 70.1% of those with vehicle treatment. Immunohistochemistry results for 8-hydroxydeoxyguanosine (OHdG) expression showed that some neurons underwent oxidative stress around the ischemic boundary zone at 6h post-ischemia. Cilostazol treatment significantly reduced the percentage of 8-OHdG-positive neurons (65.8±33.5% with vehicle and 21.3±9.9% with cilostazol). Moreover, NADPH oxidase (NOX) 2-positive neurons were significantly reduced with cilostazol treatment. In contrast, immunohistochemistry results for phosphorylated cyclic-AMP response element binding protein (pCREB) showed that there were significantly more pCREB-positive neurons around the ischemic boundary zone of cilostazol-treated mice than in those of vehicle-treated mice at 24h post-ischemia. These results suggested that cilostazol might have multiple mechanisms of action to ameliorate ischemic tissue damage, by attenuating oxidative stress mediated by suppressing NOX2 expression by ischemic neurons and an anti-apoptotic effect mediated through the pCREB pathway.
authors.affiliation: Department of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan. Electronic address: hshichi@med.hokudai.ac.jp.
authors.forename: Hideo
authors.initials: H
authors.lastname: Shichinohe
authors.affiliation: Department of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
authors.forename: Chengbo
authors.initials: C
authors.lastname: Tan
authors.affiliation: Department of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
authors.forename: Takeo
authors.initials: T
authors.lastname: Abumiya
authors.affiliation: Department of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
authors.forename: Naoki
authors.initials: N
authors.lastname: Nakayama
authors.affiliation: Department of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
authors.forename: Ken
authors.initials: K
authors.lastname: Kazumata
authors.affiliation: Department of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
authors.forename: Masaaki
authors.initials: M
authors.lastname: Hokari
authors.affiliation: Department of Neurosurgery, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
authors.forename: Kiyohiro
authors.initials: K
authors.lastname: Houkin
authors.affiliation: Department of Neurosurgery, Graduate School of Medicine and Pharmaceutical Science for Education, University of Toyama, Toyama, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Kuroda
mesh_terms: D000080242:8-Hydroxy-2'-Deoxyguanosine; D000818:Animals; D001921:Brain / Q000187:drug effects* / Q000473:pathology / Q000503:physiopathology; D002545:Brain Ischemia / Q000188:drug therapy* / Q000473:pathology / Q000503:physiopathology; D050882:CREB-Binding Protein / Q000378:metabolism; D000077407:Cilostazol; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000378:metabolism; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects / Q000473:pathology / Q000502:physiology; D007150:Immunohistochemistry; D020244:Infarction, Middle Cerebral Artery; D008297:Male; D008562:Membrane Glycoproteins / Q000378:metabolism; D008807:Mice, Inbred BALB C; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases / Q000378:metabolism; D009474:Neurons / Q000187:drug effects* / Q000473:pathology / Q000502:physiology; D018696:Neuroprotective Agents / Q000494:pharmacology*; D018384:Oxidative Stress / Q000187:drug effects / Q000502:physiology; D010766:Phosphorylation / Q000187:drug effects; D013777:Tetrazoles / Q000494:pharmacology*; D013997:Time Factors
chemical_list: D008562:Membrane Glycoproteins; D018696:Neuroprotective Agents; D013777:Tetrazoles; D000080242:8-Hydroxy-2'-Deoxyguanosine; C488842:Cybb protein, mouse; D000074662:NADPH Oxidase 2; D019255:NADPH Oxidases; D050882:CREB-Binding Protein; C494364:Crebbp protein, mouse; D003849:Deoxyguanosine; D000077407:Cilostazol
keywords: Cilostazol; NADPH oxidase 2; Oxidative stress; Permanent focal ischemia; Pleiotropic effect