A novel method using confocal laser scanning microscopy for sensitive measurement of P-glycoprotein-mediated transport activity in Caco-2 cells.

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  • Additional Information
    • Source:
      Publisher: Oxford University Press Country of Publication: England NLM ID: 0376363 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2042-7158 (Electronic) Linking ISSN: 00223573 NLM ISO Abbreviation: J Pharm Pharmacol Subsets: MEDLINE
    • Publication Information:
      Publication: 2021- : [Oxford] : Oxford University Press
      Original Publication: London : Pharmaceutical Society of Great Britain
    • Subject Terms:
      ATP Binding Cassette Transporter, Subfamily B, Member 1/*metabolism ; Biological Transport/*physiology ; Caco-2 Cells/*metabolism ; Drug Interactions/*physiology ; Microscopy, Confocal/*methods; Biological Transport/drug effects ; Digoxin/pharmacology ; Fluorescent Dyes/metabolism ; Humans ; Hypericum ; Plant Extracts/pharmacology ; Rhodamine 123/metabolism
    • Abstract:
      Objectives: The aim of this study was to use time-lapse confocal laser scanning microscopy to establish a more sensitive and specific method for evaluating P-glycoprotein activity in Caco-2 cells.
      Methods: The change in the fluorescence of residual rhodamine 123 at the apical and central regions of Caco-2 cells was measured in the presence of digoxin or St John's wort by using time-lapse confocal laser scanning microscopy. The data were compared with measurements made using conventional techniques, a fluorescence microplate reader and a fluorescence microscope.
      Key Findings: The percentage decrease of rhodamine 123 caused by 10 µm digoxin or 0.1 µg/ml St John's wort was significantly larger in the apical region of the Caco-2 cell than in the central region or in the whole cell. The digoxin-induced inhibition in the apical region as measured by time-lapse confocal laser scanning microscopy was greater than that measured in the whole cell by a microplate reader or a fluorescence microscope.
      Conclusions: The assay of residual rhodamine 123 in the apical region of Caco-2 cells by confocal laser scanning microscopy was more sensitive than the conventional methods using a microplate reader or fluorescence microscopy. It will be a valuable screening tool for studying both the inhibition and induction of P-glycoprotein activity.
      (© 2011 The Authors. JPP © 2011 Royal Pharmaceutical Society.)
    • Accession Number:
      0 (ATP Binding Cassette Transporter, Subfamily B, Member 1)
      0 (Fluorescent Dyes)
      0 (Plant Extracts)
      1N3CZ14C5O (Rhodamine 123)
      73K4184T59 (Digoxin)
    • Publication Date:
      Date Created: 20110702 Date Completed: 20120106 Latest Revision: 20181201
    • Publication Date:
      20231215
    • Accession Number:
      10.1111/j.2042-7158.2011.01294.x
    • Accession Number:
      21718284