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Naunyn-Schmiedeberg's Archives of Pharmacology

, Volume 359, Issue 4, pp 249–255 | Cite as

Functional assay of NF-κB translocation into nuclei by laser scanning cytometry: inhibitory effect by dexamethasone or theophylline

  • K. Tomita
  • Hiroki Chikumi
  • Hirokazu Tokuyasu
  • Hiroki Yajima
  • Yutaka Hitsuda
  • Yukio Matsumoto
  • Takao Sasaki
Original article

Abstract

Nuclear factor-κ B (NF-κB)/Rel transcription factors play an important role in the inducible regulation of a variety of cytokine genes in epithelial cells. We assessed accumulation of fluorescence-stained NF-κB into propi-dium iodide-stained nuclei using laser scanning cytometry. “Activity-specific” antibodies to the Rel A (p65) and NF-κB1 (p50) subunits of NF-κB were detected in the nuclei of A549 cells (an immortalized human type II alveolar epithelial cell line). Exposure to TNF-alpha caused p65 and p50 to translocate into nuclei in a dose- and time-dependent manner. Preincubation with dexamethasone for 24 h or 30 min, or with theophylline for 30 min before TNF-alpha stimulation attenuated the nuclear translocation of Rel A and p50. These findings suggest that theophylline, as well as dexamethasone, may inhibit translocation of NF-κB.

Key words Theophylline NF-kappa B Laser scanning cytometer Epithelial cell Translocation 

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Copyright information

© Springer-Verlag Berlin Heidelberg 1999

Authors and Affiliations

  • K. Tomita
    • 1
  • Hiroki Chikumi
    • 2
  • Hirokazu Tokuyasu
    • 2
  • Hiroki Yajima
    • 2
  • Yutaka Hitsuda
    • 2
  • Yukio Matsumoto
    • 2
  • Takao Sasaki
    • 2
  1. 1.Department of Thoracic Medicine, Imperial College School of Medicine at the National Heart and Lung Institute, Dovehouse Street, London, SW3 6LY, UK Fax: +44-171-3518126GB
  2. 2.Third Department of Internal Medicine, Faculty of Medicine, Tottori University, 36-1 Nishi-machi, Yonago-shi 683-8504, JapanJP

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