mRNA Expression and Amino Acid Transport Characteristics of Cultured Human Brain Microvascular Endothelial Cells (hBME)

Nobuo Umeki, Yoshiki Fukasawa, Yoshiro Kohno, Sumio Ohtsuki, Satoko Hori, Yuki Watanabe, Tetsuya Terasaki

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

An in vitro cell culture system for estimating the human blood-brain barrier (BBB) permeability of drugs is required for the development of drugs with ejects on the central nervous system. In this study, cultured human brain microvascular endothelial cells (hBME) were characterized. hBME cells exhibited concentration-dependent uptake of L-Leu, L-Glu and L-Lys with Km values of 51.1±23.1 mM, 163.3±79.8 mM and 72.4±56.6 mM, respectively. The cellular accumulation of rhodamine123 in hBME cells was unajected by P-glycoprotein (P-gp) substrates (cyclosporin A, quinidine and verapamil), while the accumulation in human P-gp-overexpressing cells was significantly increased in the presence of these P-gp substrates. RT-PCR revealed that hBME cells expressed large neutral amino acid transporter 1 (LAT1) and its associated molecule (4F2hc), excitatory amino acid transporter 3 (EAAT3), cationic amino acid transporter 1 (CAT1), glucose transporter 1 (GLUT1), monocarboxylic acid transporter 1 (MCT1) and multidrug resistance-associated protein 1 (MRP1). However, no expression of multidrug resistance protein 1 (MDR1) was detected. The results suggest that these amino acid transporters are functionally expressed at the human BBB, and that hBME cells retain the in vivo BBB transport functions and expression characteristics. Consequently, hBME cells should be a useful tool for studies of the human BBB.

Original languageEnglish
Pages (from-to)367-373
Number of pages7
JournalDrug Metabolism and Pharmacokinetics
Volume17
Issue number4
DOIs
Publication statusPublished - 2002 Jan 1
Externally publishedYes

Keywords

  • amino acid transporters
  • blood-brain barrier
  • human brain microvascular endothelial cells (hBME)
  • multidrug resistance protein 1 (MDR1)
  • multidrug resistance-associated protein 1 (MRP1)

ASJC Scopus subject areas

  • Pharmacology
  • Pharmaceutical Science
  • Pharmacology (medical)

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