title: Transendothelial permeability of chlorpyrifos in RBE4 monolayers is modulated by astrocyte-conditioned medium.
abstract: The immortalized rat brain endothelium 4 (RBE4) cell line preserves many features of the in vivo brain endothelium. It has been used as an in vitro model of the blood-brain barrier (BBB). Astrocyte-endothelial cell interactions are crucial for maintenance of BBB characteristics. The present study investigated morphological and permeability properties of the RBE4 cell line. Immunohistochemical studies showed positive staining in RBE4 cells for E-cadherin, a Ca(2+)-dependent cell-cell adhesion molecule. Western blot immunoassay showed that RBE4 cells consistently express E-cadherin and that its expression significantly increased (P<0.001) in the presence of astrocyte-conditioned medium (ACM). The transendothelial permeability of chlorpyrifos, an organophosphorus insecticide, was significantly decreased (P<0.001) when the RBE4 cells were grown in ACM compared with control medium. Additional studies were carried out to determine whether chlorpyrifos is a substrate for the multidrug resistance protein, P-glycoprotein (P-gp). No significant change in chlorpyrifos transendothelial permeability was noted in the presence of verapamil, a P-gp blocker. Thus, in this system, chlorpyrifos is not a substrate for P-gp. This work demonstrates that with additional refinements the RBE4 monolayers might serve as a useful in vitro model for the study of BBB permeability and modulation by astrocyte-derived soluble factors.
authors.affiliation: Department of Physiology and Pharmacology, Wake Forest University, School of Medicine, Winston-Salem, NC 27157, USA.
authors.forename: J
authors.initials: J
authors.lastname: Yang
authors.forename: L A
authors.initials: LA
authors.lastname: Mutkus
authors.forename: D
authors.initials: D
authors.lastname: Sumner
authors.forename: J T
authors.initials: JT
authors.lastname: Stevens
authors.forename: J C
authors.initials: JC
authors.lastname: Eldridge
authors.forename: J W
authors.initials: JW
authors.lastname: Strandhoy
authors.forename: M
authors.initials: M
authors.lastname: Aschner
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism; D000818:Animals; D001253:Astrocytes / Q000378:metabolism*; D001812:Blood-Brain Barrier / Q000187:drug effects; D001921:Brain / Q000166:cytology; D015820:Cadherins / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002461:Cell Line, Transformed / Q000187:drug effects / Q000378:metabolism; D004390:Chlorpyrifos / Q000378:metabolism*; D017077:Culture Media, Conditioned / Q000494:pharmacology; D018432:Drug Resistance, Multiple; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects*; D010539:Permeability; D051381:Rats; D014700:Verapamil / Q000494:pharmacology
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D015820:Cadherins; D002121:Calcium Channel Blockers; D017077:Culture Media, Conditioned; D014700:Verapamil; D004390:Chlorpyrifos