Cilostazol Strengthens Barrier Integrity in Brain Endothelial Cells

被引:42
作者
Horai, Shoji [1 ]
Nakagawa, Shinsuke [2 ,3 ]
Tanaka, Kunihiko [2 ]
Morofuji, Yoichi [2 ]
Couraud, Pierre-Oliver [4 ]
Deli, Maria A. [3 ,5 ]
Ozawa, Masaki [1 ]
Niwa, Masami [2 ,3 ]
机构
[1] Nagasaki Univ, Dept Neuropsychiat, Grad Sch Biomed Sci, Nagasaki 8528501, Japan
[2] Nagasaki Univ, Dept Pharmacol, Grad Sch Biomed Sci, Nagasaki 8528523, Japan
[3] PharmaCo Cell Co Ltd, BBB Lab, Nagasaki 8500862, Japan
[4] Univ Paris 05, INSERM, U1016, Inst Cochin,CNRS,UMR8104, F-75014 Paris, France
[5] Hungarian Acad Sci, Inst Biophys, Biol Res Ctr, Mol Neurobiol Lab, H-6726 Szeged, Hungary
基金
日本学术振兴会;
关键词
Cilostazol; BBB-protective drug; Phosphodiesterase; 3B; Blood-brain barrier; Tight junctions; Occludin; F-actin; P-glycoprotein; Oxygen glucose deprivation and reoxygenation; hCMEC/D3; cells; Brain endothelial cells (rat); OXYGEN-GLUCOSE DEPRIVATION; CYCLIC-NUCLEOTIDE PHOSPHODIESTERASES; P-GLYCOPROTEIN EXPRESSION; IN-VITRO; PROTEIN-KINASE; TIGHT JUNCTIONS; UP-REGULATION; INDEPENDENT PATHWAYS; OXIDATIVE STRESS; BETA-CATENIN;
D O I
10.1007/s10571-012-9896-1
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
We studied the effect of cilostazol, a selective inhibitor of phosphodiesterase 3, on barrier functions of blood-brain barrier (BBB)-related endothelial cells, primary rat brain capillary endothelial cells (RBEC), and the immortalized human brain endothelial cell line hCMEC/D3. The pharmacological potency of cilostazol was also evaluated on ischemia-related BBB dysfunction using a triple co-culture BBB model (BBB Kit (TM)) subjected to 6-h oxygen glucose deprivation (OGD) and 3-h reoxygenation. There was expression of phosphodiesterase 3B mRNA in RBEC, and a significant increase in intracellular cyclic AMP (cAMP) content was detected in RBEC treated with both 1 and 10 mu M cilostazol. Cilostazol increased the transendothelial electrical resistance (TEER), an index of barrier tightness of interendothelial tight junctions (TJs), and decreased the endothelial permeability of sodium fluorescein through the RBEC monolayer. The effects on these barrier functions were significantly reduced in the presence of protein kinase A (PKA) inhibitor H-89. Microscopic observation revealed smooth and even localization of occludin immunostaining at TJs and F-actin fibers at the cell borders in cilostazol-treated RBEC. In hCMEC/D3 cells treated with 1 and 10 mu M cilostazol for 24 and 96 h, P-glycoprotein transporter activity was increased, as assessed by rhodamine 123 accumulation. Cilostazol improved the TEER in our triple co-culture BBB model with 6-h OGD and 3-h reoxygenation. As cilostazol stabilized barrier integrity in BBB-related endothelial cells, probably via cAMP/PKA signaling, the possibility that cilostazol acts as a BBB-protective drug against cerebral ischemic insults to neurons has to be considered.
引用
收藏
页码:291 / 307
页数:17
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