title: An in vitro transport model for rapid screening and predicting the permeability of candidate compounds at blood-brain barrier.
abstract: The aim of this study was to design and develop a simple in vitro blood-brain barrier (BBB) permeation model for elementarily and rapidly predicting the permeability of candidate compounds at BBB and further evaluating whether P-glycoprotein (P-gp) affects them across BBB. The model was mainly composed of cultured rat brain microvascular endothelial cells (rBMECs), glass contraption, and micropore membrane. First, we evaluated the model by morphological observation. Second, the restriction effects of paracellular transport were verified by measuring marker probes transport, and monitoring transendothelial electrical resistance (TEER) and leakage. Finally, protein expression and activity of P-gp were confirmed by carrying out Western blot analysis and polarized transport of rhodamine-123 (Rho123) in rBMECs. The rBMECs retained both endothelial cells and BBB features. The rBMECs model reproducibly attained approximately 130 Ω cm² on the steady-state TEER value, and displayed a barrier function to marker probes transport by decreasing the permeability. Protein band of 170 kDa manifested the existence of P-gp in the rBMECs, and the findings of cyclosporin A-sensitive decrease of Rho123 efflux confirmed the presence of P-gp activity. A simple, rapid, and convenient in vitro BBB permeation model was successfully established and applied to evaluate the BBB transport profiles of three natural flavonoids: quercetin, naringenin, and rutin.
authors: Chinese Academy of Medical Sciences and Peking Union Medical College, Institute of Medicinal Plant Development, Beijing 100193, China.
authors: Zhi-Hong
authors: ZH
authors: Yang
authors: Xiao
authors: X
authors: Sun
authors: Chao
authors: C
authors: Mei
authors: Xiao-Bo
authors: XB
authors: Sun
authors: Xiao-Dong
authors: XD
authors: Liu
authors: Qi
authors: Q
authors: Chang
mesh_terms: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000378:metabolism*; D000818:Animals; D001692:Biological Transport; D001812:Blood-Brain Barrier / Q000187:drug effects / Q000378:metabolism*; D001921:Brain / Q000166:cytology / Q000378:metabolism; D044950:Flavanones / Q000494:pharmacology; D008297:Male; D008954:Models, Biological; D011794:Quercetin / Q000494:pharmacology; D051381:Rats; D020112:Rhodamine 123 / Q000494:pharmacology*; D012431:Rutin / Q000494:pharmacology
chemical_list: D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D044950:Flavanones; D020112:Rhodamine 123; D012431:Rutin; D011794:Quercetin; C005273:naringenin