Cilostazol strengthens barrier integrity in brain endothelial cells

Shoji Horai, Shinsuke Nakagawa, Kunihiko Tanaka, Yoichi Morofuji, Pierre Oliver Couraud, M. Deli, Masaki Ozawa, Masami Niwa

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

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™) 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 μM 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 μM 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.

Original languageEnglish
Pages (from-to)291-307
Number of pages17
JournalCellular and Molecular Neurobiology
Volume33
Issue number2
DOIs
Publication statusPublished - Mar 2013

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Blood-Brain Barrier
Endothelial Cells
Brain
Tight Junctions
Coculture Techniques
Cyclic AMP-Dependent Protein Kinases
Electric Impedance
Cyclic AMP
Phosphodiesterase 3 Inhibitors
Type 3 Cyclic Nucleotide Phosphodiesterases
Oxygen
Occludin
Rhodamine 123
cilostazol
Protective Agents
Glucose
Adenylate Kinase
P-Glycoprotein
Protein Kinase Inhibitors
Fluorescein

Keywords

  • BBB-protective drug
  • Blood-brain barrier
  • Brain endothelial cells (rat)
  • Cilostazol
  • F-actin
  • hCMEC/D3 cells
  • Occludin
  • Oxygen glucose deprivation and reoxygenation
  • P-glycoprotein
  • Phosphodiesterase 3B
  • Tight junctions

ASJC Scopus subject areas

  • Cellular and Molecular Neuroscience
  • Cell Biology

Cite this

Horai, S., Nakagawa, S., Tanaka, K., Morofuji, Y., Couraud, P. O., Deli, M., ... Niwa, M. (2013). Cilostazol strengthens barrier integrity in brain endothelial cells. Cellular and Molecular Neurobiology, 33(2), 291-307. https://doi.org/10.1007/s10571-012-9896-1

Cilostazol strengthens barrier integrity in brain endothelial cells. / Horai, Shoji; Nakagawa, Shinsuke; Tanaka, Kunihiko; Morofuji, Yoichi; Couraud, Pierre Oliver; Deli, M.; Ozawa, Masaki; Niwa, Masami.

In: Cellular and Molecular Neurobiology, Vol. 33, No. 2, 03.2013, p. 291-307.

Research output: Contribution to journalArticle

Horai, S, Nakagawa, S, Tanaka, K, Morofuji, Y, Couraud, PO, Deli, M, Ozawa, M & Niwa, M 2013, 'Cilostazol strengthens barrier integrity in brain endothelial cells', Cellular and Molecular Neurobiology, vol. 33, no. 2, pp. 291-307. https://doi.org/10.1007/s10571-012-9896-1
Horai, Shoji ; Nakagawa, Shinsuke ; Tanaka, Kunihiko ; Morofuji, Yoichi ; Couraud, Pierre Oliver ; Deli, M. ; Ozawa, Masaki ; Niwa, Masami. / Cilostazol strengthens barrier integrity in brain endothelial cells. In: Cellular and Molecular Neurobiology. 2013 ; Vol. 33, No. 2. pp. 291-307.
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