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Molecular Pharmacology

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Research ArticleArticle

Analysis of In Vivo Nuclear Factor-κB Activation during Liver Inflammation in Mice: Prevention by Catalase Delivery

Kenji Hyoudou, Makiya Nishikawa, Yuki Kobayashi, Yukari Kuramoto, Fumiyoshi Yamashita and Mitsuru Hashida
Molecular Pharmacology February 2007, 71 (2) 446-453; DOI: https://doi.org/10.1124/mol.106.027169
Kenji Hyoudou
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Makiya Nishikawa
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Yuki Kobayashi
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Yukari Kuramoto
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Fumiyoshi Yamashita
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Mitsuru Hashida
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Abstract

Nuclear factor-κB (NF-κB) is a transcription factor that plays crucial roles in inflammation, immunity, cell proliferation, and apoptosis. Until now, there have been few studies of NF-κB activation in whole animals because of experimental difficulties. Here, we show that mice receiving a simple injection of plasmid vectors can be used to examine NF-κB activation in the liver. Two plasmid vectors, pNF-κB-Luc (firefly luciferase gene) and pRL-SV40 (Renilla reniformis luciferase gene), were injected into the tail vein of mice by the hydrodynamics-based procedure, an established method of gene transfer to mouse liver. Then, the ratio of the firefly and R. reniformis luciferase activities (F/R) was used as an indicator of the NF-κB activity in the liver. Injection of thioacetamide or lipopolysaccharide plus d-galactosamine increased the F/R ratio in the liver, and this was significantly (P < 0.001) inhibited by an intravenous injection of catalase derivatives targeting liver nonparenchymal cells. Imaging the firefly luciferase expression in live mice clearly demonstrated that the catalase derivatives efficiently prevented the NF-κB-mediated expression of the firefly luciferase gene. Plasma transaminases and the survival rate of mice supported the findings obtained by the luminescence-based analyses. Thus, this method, which requires no genetic recombination techniques, is highly sensitive to the activation of NF-κB and allows us to continuously examine the activation in live animals. In conclusion, this novel, simple, and sensitive method can be used not only for analyzing the NF-κB activation in the organ under different inflammatory conditions but also for screening drug candidates for the prevention of liver inflammation.

Footnotes

  • This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan, by Health and Labor Sciences Research Grants for Research on Hepatitis and BSE from the Ministry of Health, Labor and Welfare of Japan and by the 21st Century COE Program “Knowledge Information Infrastructure for Genome Science.”

  • ABBREVIATIONS: TNF, tumor necrosis factor; Gal-catalase, galactosylated catalase; Man-catalase, mannosylated catalase; PEG-catalase, polyethyleneglycol-conjugated catalase; Suc-catalase, succinylated catalase; TA, thioacetamide; LPS, lipopolysaccharide; GalN, d-galactosamine; IL, interleukin; ROS, reactive oxygen species; EMSA, electrophoretic mobility shift assay; NF-κB, nuclear factor-κB; F/R, ratio of the firefly and Renilla reniformis luciferase activities; AST, aspartate aminotransferase; ALT, alanine aminotransferase; SV40, simian virus 40.

  • ↵ Embedded Image The online version of this article (available at http://molpharm.aspetjournals.org) contains supplemental material.

    • Received May 25, 2006.
    • Accepted November 14, 2006.
  • The American Society for Pharmacology and Experimental Therapeutics
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Molecular Pharmacology: 71 (2)
Molecular Pharmacology
Vol. 71, Issue 2
1 Feb 2007
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Research ArticleArticle

Analysis of In Vivo Nuclear Factor-κB Activation during Liver Inflammation in Mice: Prevention by Catalase Delivery

Kenji Hyoudou, Makiya Nishikawa, Yuki Kobayashi, Yukari Kuramoto, Fumiyoshi Yamashita and Mitsuru Hashida
Molecular Pharmacology February 1, 2007, 71 (2) 446-453; DOI: https://doi.org/10.1124/mol.106.027169

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Research ArticleArticle

Analysis of In Vivo Nuclear Factor-κB Activation during Liver Inflammation in Mice: Prevention by Catalase Delivery

Kenji Hyoudou, Makiya Nishikawa, Yuki Kobayashi, Yukari Kuramoto, Fumiyoshi Yamashita and Mitsuru Hashida
Molecular Pharmacology February 1, 2007, 71 (2) 446-453; DOI: https://doi.org/10.1124/mol.106.027169
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