title: Effect of pluronic p85 on amino acid transport in bovine brain microvessel endothelial cells.
abstract: A synthetic amphiphilic block copolymer Pluronic P85 (P85) was shown to be among the most potent inhibitors of Pgp efflux system in the blood-brain barrier (BBB) and capable of enhancing delivery of Pgp substrates to the brain. The purpose of this work is to evaluate the effects of P85 on amino acid transport in BBB. Primary bovine brain microvessel endothelial cells (BBMEC) grown on membrane inserts were used as an in vitro BBB model. Expression of amino acid transporters, like large neutral amino acid transporter 1, cationic amino acid transporter 1, and small neutral amino acid transporter 1, were confirmed by reverse transcriptase polymerase chain reaction. Effects of P85 on amino acid transporters were examined using their substrates: (3)H-phenylalanine, (3)H-lysine, and (3)H-methylaminoisobutyric acid, respectively. BBMEC permeability studies were carried out in apical (AP) to basolateral (BL) and BL to AP directions. P85 added at the AP side had little, if any, effect on AP to BL ("blood to brain") transport for all examined amino acids in BBMEC monolayers. However, 0.1% P85 added at the BL side significantly increased the BL to AP transport of these substrates. Furthermore, the effective concentrations of P85 were also shown to induce plasma membrane depolarization and increase intracellular sodium concentration in BBMEC, which can contribute to the effects of the copolymer on the energy-dependent transport systems. All together, despite profound effects on transport system(s) at the brain side of cell monolayers, P85 had no effect on AP to BL transport of amino acids in brain microvessel endothelial cell model.
authors.affiliation: Department of Pharmaceutical Sciences and Center for Drug Delivery and Nanomedicine, College of Pharmacy, University of Nebraska Medical Center, Omaha, NE 68198, USA.
authors.forename: Xiaobin
authors.initials: X
authors.lastname: Zhang
authors.forename: Daria Y
authors.initials: DY
authors.lastname: Alakhova
authors.forename: Elena V
authors.initials: EV
authors.lastname: Batrakova
authors.forename: Shu
authors.initials: S
authors.lastname: Li
authors.forename: Zhihui
authors.initials: Z
authors.lastname: Yang
authors.forename: Yili
authors.initials: Y
authors.lastname: Li
authors.forename: Alexander V
authors.initials: AV
authors.lastname: Kabanov
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism; D026905:Amino Acid Transport Systems / Q000378:metabolism*; D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D001693:Biological Transport, Active / Q000187:drug effects; D001923:Brain Chemistry / Q000187:drug effects*; D002196:Capillaries / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D002417:Cattle; D002469:Cell Separation; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D008564:Membrane Potentials / Q000187:drug effects; D018613:Microscopy, Confocal; D011060:Poloxalene / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D026905:Amino Acid Transport Systems; D000596:Amino Acids; C422648:pluronic block copolymer p85; D011060:Poloxalene