Serum-derived factors weaken the barrier properties of cultured porcine brain capillary endothelial cells in vitro

Nitz, Thorsten; Eisenblätter, Tanja; Psathaki, Katherina; Galla, Hans‐Joachim

doi:10.1016/s0006-8993(03)02834-8

Cited by 70 publications

(57 citation statements)

References 51 publications

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“…We also observed the reduction in TEER of the control BBB Kit TM used in normoxia experiments (93 ± 9 Ω × cm 2 , n = 8). As the same phenomenon was previously reported by Nitz et al (2003), we thought that the reduction was due to the medium change in culture that in normoxia experiments we changed the medium the BBB Kit medium containing 10 % (v/v) PDS to serum-free DEMN containing 4.5 g/dl glucose (glucose + DMEM).…”

Section: Resultssupporting

confidence: 58%

Cilostazol Strengthens Barrier Integrity in Brain Endothelial Cells

et al. 2012

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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.

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Section: Resultssupporting

confidence: 58%

Cilostazol Strengthens Barrier Integrity in Brain Endothelial Cells

et al. 2012

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“…The effects of LPA on vascular permeability are a topic of considerable debate with some studies reporting a decrease in endothelial cell permeability after LPA exposure, 14,15 whereas others support an increase in vascular leakage after LPA exposure. 11,12,16 Studies of LPA on brain microvessel endothelial cell permeability support an enhancement of permeability with decrease in transcellular electrical resistance and increases in flux of vascular markers observed in primary cultured brain endothelial cell preparations. 12,16 More recent studies indicate increased permeability after localized application of LPA to the cerebral microvasculature.…”

Section: Introductionmentioning

confidence: 82%

“…12,16 In cultured BMECs, LPA exposure has been associated with increases in permeability. 11,12 However, there are contradictory reports of LPA-induced reductions in endothelial cell permeability (i.e., barrier-enhancing properties). 15 The conflicting reports regarding LPA-induced permeability could be dependent on a number of factors including differences in vascular bed, species, or the profile of LPA receptors expressed.…”

Section: Studies By Yanagida K Et Almentioning

confidence: 99%

Rapid and Reversible Enhancement of Blood–Brain Barrier Permeability Using Lysophosphatidic Acid

Savant

Toews

et al. 2013

J Cereb Blood Flow Metab

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The present study characterizes the effects of lysophosphatidic acid (LPA) on blood-brain barrier (BBB) permeability focusing specifically on the time of onset, duration, and magnitude of LPA-induced changes in cerebrovascular permeability in the mouse using both magnetic resonance imaging (MRI) and near infrared fluorescence imaging (NIFR). Furthermore, potential application of LPA for enhanced drug delivery to the brain was also examined by measuring the brain accumulation of radiolabeled methotrexate. Exposure of primary cultured brain microvessel endothelial cells (BMECs) to LPA produced concentration-dependent increases in permeability that were completely abolished by clostridium toxin B. Administration of LPA disrupted BBB integrity and enhanced the permeability of small molecular weight marker gadolinium diethylenetriaminepentaacetate (Gd-DTPA) contrast agent, the large molecular weight permeability marker, IRdye800cwPEG, and the P-glycoprotein efflux transporter probe, Rhodamine 800 (R800). The increase in BBB permeability occurred within 3 minutes after LPA injection and barrier integrity was restored within 20 minutes. A decreased response to LPA on large macromolecule BBB permeability was observed after repeated administration. The administration of LPA also resulted in 20-fold enhancement of radiolabeled methotrexate in the brain. These studies indicate that administration of LPA in combination with therapeutic agents may increase drug delivery to the brain.

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“…The localization of the TJ proteins within the endothelium is an important factor that also influences the permeability after treatment with glucocorticoids (7,35). Therefore, although decreases in barrier permeability showed correlations with increases in the expression of some of the TJ proteins, localization of the TJ proteins to periphery of cell borders also contributes to increases in transendothelial electrical resistances and decreases in permeability (7,35).…”

Section: Discussionmentioning

confidence: 99%

“…Positive controls were used on the same immunoblot with the sheep brain samples to establish with certainty that the proteins that were identified by these antibodies in the sheep brain were the same as the known protein for these specific antibodies. Detection of the occludin, claudin-1, claudin-5, JAM-A, ZO-1, and ZO-2 bands at 65, 23,23,35,225, and 160 kDa, respectively, was dependent on incubation with the primary antibody, the omission of which resulted in the absence of this signal.…”

Section: Methodsmentioning

confidence: 99%

Maternal glucocorticoid exposure alters tight junction protein expression in the brain of fetal sheep

Sadowska

Malaeb

Stonestreet

2010

American Journal of Physiology-Heart and Circulatory Physiology

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Sadowska GB, Malaeb SN, Stonestreet BS. Maternal glucocorticoid exposure alters tight junction protein expression in the brain of fetal sheep. Am J Physiol Heart Circ Physiol 298: H179 -H188, 2010. First published October 23, 2009 doi:10.1152/ajpheart.00828.2009.-We examined the expression of tight junction (TJ) proteins in the cerebral cortex, cerebellum, and spinal cord of fetuses after maternal treatment with single and multiple courses of dexamethasone. Ewes received either single courses of four 6-mg dexamethasone or placebo injections every 12 h for 48 h between 104 and 107 days or the same treatment once a week between 76 -78 and 104 -107 days of gestation. TJ protein expression was determined by Western immunoblot analysis on tissue harvested at 105-108 days of gestation. Blood-brain barrier permeability has been previously quantified with the blood-tobrain transfer constant (K i) with ␣-aminoisobutyric acid (39). After a single course of dexamethasone, claudin-5 increased (P Ͻ 0.05) in the cerebral cortex, occludin and claudin-1 increased in the cerebellum, and occludin increased in the spinal cord. After multiple dexamethasone courses, occludin and zonula occludens (ZO)-1 increased in the cerebral cortex, and occludin and claudin-1 increased in the cerebellum. Junctional adhesion molecule-A and ZO-2 expressions did not change. Linear regression comparing K i to TJ proteins showed inverse correlations with claudin-1 and claudin-5 in the cerebral cortex after a single course and ZO-2 in the spinal cord after multiple courses and direct correlations with ZO-1 in the cerebellum and spinal cord after multiple courses. We conclude that maternal glucocorticoid treatment increases the expression of specific TJ proteins in vivo, patterns of TJ protein expression vary after exposure to single and multiple glucocorticoid courses, and decreases in blood-brain barrier permeability are associated with increases in claudin-1, claudin-5, and ZO-2 expression and decreases in ZO-1 expression. In utero glucocorticoid exposure alters the molecular composition of the barrier and affects fetal blood-brain barrier function.blood-brain barrier; dexamethasone; steroids THE BLOOD-BRAIN BARRIER is composed of a continuous layer of cerebrovascular endothelial cells connected by intercellular tight junctions (TJs) (4). TJs are composed of transmembrane and associated cytoplasmic proteins (22). The transmembrane proteins include occludin, claudins, and junctional adhesion molecules (JAMs). The proteins of the claudin family form the primary seal between adjacent endothelial cells, whereas occludin is an important support molecule that increases electrical resistance across the barrier and decreases paracellular permeability (22). JAMs regulate TJ formation and leukocyte and endothelial cell interactions (13). The associated cytoplasmic proteins zonula occuldens (ZO)-1 and ZO-2 stabilize TJs by connecting them to actin (22).Maternally administered glucocorticoids have been widely used to accelerate fetal maturation (9,12,30,34). Th...

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Serum-derived factors weaken the barrier properties of cultured porcine brain capillary endothelial cells in vitro

Cited by 70 publications

References 51 publications

Cilostazol Strengthens Barrier Integrity in Brain Endothelial Cells

Cilostazol Strengthens Barrier Integrity in Brain Endothelial Cells

Rapid and Reversible Enhancement of Blood–Brain Barrier Permeability Using Lysophosphatidic Acid

Maternal glucocorticoid exposure alters tight junction protein expression in the brain of fetal sheep

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