Degree grantor/typeNagasaki University (長崎大学)
Note (General)Background: Cerebral infarction accounts for 85% of all stroke cases. Even in an era of rapid and effective recanalization using an intravascular approach, the majority of patients have poor functional outcomes. Thus, there is an urgent need for the development of therapeutic agents to treat acute ischemic stroke. We evaluated the effect of fasudil, a Rho kinase inhibitor, on blood brain barrier (BBB) functions under normoxia or oxygen–glucose deprivation (OGD) conditions using a primary cell-based in vitro BBB model.Methods: BBB models from rat primary cultures (brain capillary endothelial cells, astrocytes, and pericytes) were subjected to either normoxia or 6 h OGD/24 h reoxygenation. To assess the effects of fasudil on BBB functions, we evaluated real time impedance, transendothelial electrical resistance (TEER), sodium fluorescein permeability, and tight junction protein expression using western blotting. Lastly, to understand the observed protective mechanism on BBB functions by fasudil we examined the role of cyclooxygenase-2 and thromboxane A2 receptor agonist U-46619 in BBB-forming cells.Results: We found that treatment with 0.3–30 µM of fasudil increased cellular impedance. Fasudil enhanced barrier properties in a concentration-dependent manner, as measured by an increased (TEER) and decreased permeability. Fasudil also increased the expression of tight junction protein claudin-5. Reductions in TEER and increased permeability were observed after OGD/reoxygenation exposure in mono- and co-culture models. The improvement in BBB integrity by fasudil was confirmed in both of the models, but was significantly higher in the co-culture than in the monoculture model. Treatment with U-46619 did not show significant changes in TEER in the monoculture model, whereas it showed a significant reduction in TEER in the co-culture model. Fasudil significantly improved the U-46619-induced TEER reduction in the co-culture models. Pericytes and astrocytes have opposite effects on endothelial cells and may contribute to endothelial injury in hyperacute ischemic stroke. Overall, fasudil protects the integrity of BBB both by a direct protective effect on endothelial cells and by a pathway mediated via pericytes and astrocytes.Conclusions: Our findings suggest that fasudil is a BBB-protective agent against acute ischemic stroke.
長崎大学学位論文 学位記番号:博(医歯薬)甲第1473号 学位授与年月日:令和4年12月7日
Author: Kei Sato, Shinsuke Nakagawa, Yoichi Morofuji, Yuki Matsunaga, Takashi Fujimoto, Daisuke Watanabe, Tsuyoshi Izumo, Masami Niwa, Fruzsina R. Walter, Judit P. Vigh, Ana Raquel Santa-Maria, Maria A. Deli & Takayuki Matsuo
Citation: Fluids and Barriers of the CNS, 19, art.no. 43; 2022
identifier:Nagasaki University (長崎大学), 博士(医学) (2022-12-07)
http://hdl.handle.net/10069/00041980
Collection (particular)国立国会図書館デジタルコレクション > デジタル化資料 > 博士論文
Date Accepted (W3CDTF)2023-03-07T16:07:52+09:00
Data Provider (Database)国立国会図書館 : 国立国会図書館デジタルコレクション