Advertisement

Inflammation Research

, Volume 51, Issue 4, pp 188–194 | Cite as

An anti-rheumatic agent T-614 inhibits NF-κB activation in LPS- and TNF-α-stimulated THP-1 cells without interfering with IκBα degradation

  • Y. Aikawa
  • M. Yamamoto
  • T. Yamamoto
  • K. Morimoto
  • K. Tanaka

Abstract.

Objective: Compound T-614, a member of the methanesulfoanilide class of anti-inflammatory agents, shows potent anti-arthritic activity in animal models of rheumatoid arthritis. The aim of the present investigation was to characterize the anti-arthritic activity of T-614 in terms of regulation of the nuclear transcription factor NF-κB, which is associated with expression of many immune and inflammatory genes.¶Materials and methods: THP-1 cells (human monocytic leukemia cell line) were used throughout this in vitro study, and lipopolysaccharide (LPS) and tumor necrosis factor (TNF)- α were employed for activation of the cells. Cytokine production was assayed by enzyme-linked immunosorbent assay (ELISA). The mRNA levels were determined by a semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) assay. Assessment of the NF-κB DNA binding activity was performed by an electrophoretic mobility shift assay (EMSA) using a digoxigenin (DIG)-labeled double-stranded oligonucleotide containing κB-binding site. Degradation kinetics of the cytosolic NF-κB inhibitor α (IκBα) were studied by Western blot analysis.¶Results: T-614 inhibited LPS-stimulated production of TNF-α, interleukin (IL)-6, and IL-8 in a concentration-dependent manner with decreasing mRNA levels (IL-6 and IL-8). EMSA study showed that T-614 prevented TNF-α as well as LPS-stimulated activation of NF-κB, and Western blot analysis proved that T-614 did not affect degradation of IκBα protein.¶Conclusions: These results suggest that the inhibitory effect of T-614 on the production of TNF-α, IL-6 and IL-8 in LPS-stimulated THP-1 cells may involve transcriptional regulation through suppression of NF-κB activation without interfering with IκBα degradation.

Key words: Cytokines - T-614 - NF-κB - IκBα - Anti-rheumatic agent 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Copyright information

© Birkhäuser Verlag, 2002

Authors and Affiliations

  • Y. Aikawa
    • 1
  • M. Yamamoto
    • 1
  • T. Yamamoto
    • 1
  • K. Morimoto
    • 1
  • K. Tanaka
    • 1
  1. 1.Research Laboratories, Toyama Chemical Co., Ltd., Shimookui 2-4-1, Toyama 930-8508, Japan, Fax: ++ 81 76 431 8208, e-mail: yukihiko_aikawa@toyama-chemical.co.jp JP

Personalised recommendations