Kinetic analysis of P-glycoprotein-mediated transport by using normal human placental brush-border membrane vesicles

Fumihiko Ushigome, Noriko Koyabu, Shoji Satoh, Kiyomi Tsukimori, Hitoo Nakano, Takanori Nakamura, Takeshi Uchiumi, Michihiko Kuwano, Hisakazu Ohtani, Yasufumi Sawada

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Purpose. P-Glycoprotein (Pgp) plays an important role in drug disposition and excretion in various tissues such as the brain, intestine, and kidney. Moreover, we have demonstrated that Pgp is expressed on the brush-border membranes of trophoblast cells in the placenta and restricts drug transfer from the maternal circulation to the fetus. However, the transport kinetics of physiologically expressed Pgp has scarcely been investigated. Methods. In this study, we assessed the functional kinetics of transport mediated by Pgp that is physiologically expressed in normal tissue by using human placental brush-border membrane vesicles (BBMVs). Digoxin and vinblastine were used as typical substrates of Pgp. Results. The uptakes of [3H] digoxin and [3H] vinblastine into BBMVs were significantly increased in the presence of an ATP-regenerating system. The ATP-dependent uptakes of [3H]digoxin and [3H] vinblastine into BBMVs exhibited saturable kinetics. The Michaelis constants (Kt values) were 2.65 ± 1.80 μM and 21.9 ± 3.37 μM, respectively. In the presence of a Pgp inhibitor such as verapamil, cyclosporine A, or progesterone, the ATP-dependent uptakes of [3H]digoxin and [3H] vinblastine into BBMVs were significantly reduced. Anti-Pgp monoclonal antibody C219 completely inhibited the uptake of [3H] digoxin. Conclusions. The transport kinetics of [3H]digoxin and [3H]vinblastine by physiologically expressed Pgp were successfully evaluated by using BBMVs prepared from normal human placenta. The present method enabled us to evaluate the function of physiologically expressed Pgp and is superior to the use of cultured transfectants in terms of the yield of vesicles. The present method may also be applicable to investigating the influence of various factors such as the genotype of the MDR1 gene or various pathophysiologic states of neonates on the function of Pgp.

Original languageEnglish
Pages (from-to)38-44
Number of pages7
JournalPharmaceutical Research
Volume20
Issue number1
DOIs
Publication statusPublished - 2003 Jan 1
Externally publishedYes

Fingerprint

P-Glycoprotein
Brushes
Microvilli
Digoxin
Membranes
Kinetics
Vinblastine
Adenosine Triphosphate
Placenta
Tissue
Trophoblasts
Verapamil
Pharmaceutical Preparations
Cyclosporine
Intestines
Progesterone
Brain
Fetus
Genes
Genotype

Keywords

  • Blood-placental barrier
  • Brush-border membrane vesicles
  • Digoxin
  • P-glycoprotein
  • Vinblastine

ASJC Scopus subject areas

  • Chemistry(all)
  • Pharmaceutical Science
  • Pharmacology

Cite this

Ushigome, F., Koyabu, N., Satoh, S., Tsukimori, K., Nakano, H., Nakamura, T., ... Sawada, Y. (2003). Kinetic analysis of P-glycoprotein-mediated transport by using normal human placental brush-border membrane vesicles. Pharmaceutical Research, 20(1), 38-44. https://doi.org/10.1023/A:1022290523347

Kinetic analysis of P-glycoprotein-mediated transport by using normal human placental brush-border membrane vesicles. / Ushigome, Fumihiko; Koyabu, Noriko; Satoh, Shoji; Tsukimori, Kiyomi; Nakano, Hitoo; Nakamura, Takanori; Uchiumi, Takeshi; Kuwano, Michihiko; Ohtani, Hisakazu; Sawada, Yasufumi.

In: Pharmaceutical Research, Vol. 20, No. 1, 01.01.2003, p. 38-44.

Research output: Contribution to journalArticle

Ushigome, F, Koyabu, N, Satoh, S, Tsukimori, K, Nakano, H, Nakamura, T, Uchiumi, T, Kuwano, M, Ohtani, H & Sawada, Y 2003, 'Kinetic analysis of P-glycoprotein-mediated transport by using normal human placental brush-border membrane vesicles', Pharmaceutical Research, vol. 20, no. 1, pp. 38-44. https://doi.org/10.1023/A:1022290523347
Ushigome, Fumihiko ; Koyabu, Noriko ; Satoh, Shoji ; Tsukimori, Kiyomi ; Nakano, Hitoo ; Nakamura, Takanori ; Uchiumi, Takeshi ; Kuwano, Michihiko ; Ohtani, Hisakazu ; Sawada, Yasufumi. / Kinetic analysis of P-glycoprotein-mediated transport by using normal human placental brush-border membrane vesicles. In: Pharmaceutical Research. 2003 ; Vol. 20, No. 1. pp. 38-44.
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AU - Koyabu, Noriko

AU - Satoh, Shoji

AU - Tsukimori, Kiyomi

AU - Nakano, Hitoo

AU - Nakamura, Takanori

AU - Uchiumi, Takeshi

AU - Kuwano, Michihiko

AU - Ohtani, Hisakazu

AU - Sawada, Yasufumi

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KW - P-glycoprotein

KW - Vinblastine

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