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

20 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

Fingerprint

Endothelial Cells
Amino Acids
Messenger RNA
Brain
P-Glycoprotein
Blood-Brain Barrier
Excitatory Amino Acid Transporter 3
Large Neutral Amino Acid-Transporter 1
Cationic Amino Acid Transporter 1
Monocarboxylic Acid Transporters
Amino Acid Transport Systems
Facilitative Glucose Transport Proteins
Pharmaceutical Preparations
Cyclosporine
Permeability
Central Nervous System
Cell Culture Techniques
Polymerase Chain Reaction

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)

Cite this

mRNA Expression and Amino Acid Transport Characteristics of Cultured Human Brain Microvascular Endothelial Cells (hBME). / Umeki, Nobuo; Fukasawa, Yoshiki; Kohno, Yoshiro; Ohtsuki, Sumio; Hori, Satoko; Watanabe, Yuki; Terasaki, Tetsuya.

In: Drug Metabolism and Pharmacokinetics, Vol. 17, No. 4, 01.01.2002, p. 367-373.

Research output: Contribution to journalArticle

Umeki, Nobuo ; Fukasawa, Yoshiki ; Kohno, Yoshiro ; Ohtsuki, Sumio ; Hori, Satoko ; Watanabe, Yuki ; Terasaki, Tetsuya. / mRNA Expression and Amino Acid Transport Characteristics of Cultured Human Brain Microvascular Endothelial Cells (hBME). In: Drug Metabolism and Pharmacokinetics. 2002 ; Vol. 17, No. 4. pp. 367-373.
@article{4305ba7f821e45cb92685ea2eca6f6e6,
title = "mRNA Expression and Amino Acid Transport Characteristics of Cultured Human Brain Microvascular Endothelial Cells (hBME)",
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.",
keywords = "amino acid transporters, blood-brain barrier, human brain microvascular endothelial cells (hBME), multidrug resistance protein 1 (MDR1), multidrug resistance-associated protein 1 (MRP1)",
author = "Nobuo Umeki and Yoshiki Fukasawa and Yoshiro Kohno and Sumio Ohtsuki and Satoko Hori and Yuki Watanabe and Tetsuya Terasaki",
year = "2002",
month = "1",
day = "1",
doi = "10.2133/dmpk.17.367",
language = "English",
volume = "17",
pages = "367--373",
journal = "Drug Metabolism and Pharmacokinetics",
issn = "1347-4367",
publisher = "Japanese Society for the Study of Xenobiotics",
number = "4",

}

TY - JOUR

T1 - mRNA Expression and Amino Acid Transport Characteristics of Cultured Human Brain Microvascular Endothelial Cells (hBME)

AU - Umeki, Nobuo

AU - Fukasawa, Yoshiki

AU - Kohno, Yoshiro

AU - Ohtsuki, Sumio

AU - Hori, Satoko

AU - Watanabe, Yuki

AU - Terasaki, Tetsuya

PY - 2002/1/1

Y1 - 2002/1/1

N2 - 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.

AB - 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.

KW - amino acid transporters

KW - blood-brain barrier

KW - human brain microvascular endothelial cells (hBME)

KW - multidrug resistance protein 1 (MDR1)

KW - multidrug resistance-associated protein 1 (MRP1)

UR - http://www.scopus.com/inward/record.url?scp=85013697845&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85013697845&partnerID=8YFLogxK

U2 - 10.2133/dmpk.17.367

DO - 10.2133/dmpk.17.367

M3 - Article

VL - 17

SP - 367

EP - 373

JO - Drug Metabolism and Pharmacokinetics

JF - Drug Metabolism and Pharmacokinetics

SN - 1347-4367

IS - 4

ER -