Nuclear factor kappaB (NF-kappaB) activation by hydrogen peroxide in human epidermal keratinocytes and the restorative effect of interleukin-10.
The heterodimeric form of nuclear factor kappaB (NF-kappaB), NF-kappaB1/RelA, is one of the pluripotential transcription factors that activates various genes encoding cytokines and cell adhesion molecules. To clarify the involvement of radical oxygen species in the NF-kappaB activation pathway in keratinocytes, we examined the effect of hydrogen peroxide (H(2)O(2)) on the activation of NF-kappaB in cultured normal human epidermal keratinocytes. After the treatment of keratinocytes with 300 microM H(2)O(2), a translocation of NF-kappaB from the cytoplasm to nucleus was observed in an immunofluorescence study using anti-human NF-kappaB1 and anti-human RelA antibodies. Specific DNA binding was observed with the nuclear extract prepared from the H(2)O(2)-treated keratinocytes by the electrophoretic mobility shift assay. The presence of N-acetyl-L-cysteine or pyrrolidine dithiocarbamate during H(2)O(2) treatment prevented the nuclear localization of NF-kappaB. The involvement of radical oxygen species in the NF-kappaB activation pathway was suggested. Pretreatment of keratinocytes with 10 ng/ml of recombinant human interleukin-10 (IL-10) for 24 h suppressed the nuclear translocation of NF-kappaB induced by H(2)O(2). IL-10, which increases in ultraviolet-irradiated skin and suppresses delayed type hypersensitivity in vivo, may play an inhibitory role in cutaneous inflammation by inhibiting the NF-kappaB activation pathway in keratinocytes.
Ikeda
Mistunori
M
Department of Dermatology, Kochi Medical School, Nankoku, 783-8505, Kochi, Japan. ikedamit@kochi-ms.ac.jp
Hirose
Yasuaki
Y
Miyoshi
Ken
K
Kodama
Hajime
H
D000111:Acetylcysteine / Q000494:pharmacology; D021581:Active Transport, Cell Nucleus / Q000187:drug effects; D000975:Antioxidants / Q000494:pharmacology; D003593:Cytoplasm / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D016753:Interleukin-10 / Q000494:pharmacology*; D015603:Keratinocytes / Q000187:drug effects* / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism*; D052002:NF-kappa B p50 Subunit; D017382:Reactive Oxygen Species / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology; D051996:Transcription Factor RelA
D000975:Antioxidants; D016328:NF-kappa B; D052002:NF-kappa B p50 Subunit; D017382:Reactive Oxygen Species; D011994:Recombinant Proteins; D051996:Transcription Factor RelA; D016753:Interleukin-10; D006861:Hydrogen Peroxide; D000111:Acetylcysteine