Cilostazol Strengthens Barrier Integrity in Brain Endothelial Cells

Abstract

We studied the effect of cilostazol, a selective inhibitor of phosphodiesterase 3 (PDE3), 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 cAMP content was detected in RBEC treated with both 1 µM 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 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.

PACAP Enhances Barrier Properties of Cerebral Microvessels

“…Intracellular cAMP is known to strengthen the barrier function of the cerebral endothelium (Stelzner et al 1989 ;Rubin et al 1991 ;Rubin and Staddon 1999 ;Ishizaki et al 2003 ). Moreover, cAMP-elevating agents, like adrenomedullin or cilostazol have also been shown to stabilize the tight junctions of cerebral endothelial cells (Kis et al 2001 ;Horai et al 2013 ). Nevertheless, all three receptors of PACAP (PAC1, VPAC1, and VPAC2) signal mainly through the cAMP/PKA pathway.…”

Phosphodiesterase Inhibitors as Therapeutics for Traumatic Brain Injury

“…Relevant to the above is that the dual-specific PDE, PDE3, is strongly expressed in endothelial cells and the PDE3-selective inhibitor, cilostazol, appears to strengthens barrier integrity in brain endothelial cells [110, 111]. Furthermore, in ischemia-reperfusion injury, PDE3B levels increase in neurons and astrocytes, with activation in the latter being stronger [110].…”

Hepatocyte growth factor enhances the barrier function in primary cultures of rat brain microvascular endothelial cells

“…The flux of sodium fluorescein (Na-F) and Evans blue-albumin (EBA) across the endothelial monolayer was determined as previously described (Nakagawa et al 2007;Horai et al 2013). Na-F and EBA were used for permeability markers of small molecule and large molecule, respectively.…”