Signaling role of intracellular iron in NF-κb activation

Shigang Xiong, Hongyun She, Heigo Takeuchi, Bora Han, John F. Engelhardt, C. H. Barton, Ebrahim Zandi, Cecilia R Giulivi, Hidekazu Tsukamoto

Research output: Contribution to journalArticle

125 Scopus citations

Abstract

Iron chelators inhibit endotoxin-induced NF-κB activation in hepatic macrophages (HMs), suggesting a role for the intracellular chelatable pool of iron in NF-κB activation. The present study tested this hypothesis. Analysis of Fe59-loaded HMs stimulated with lipopolysaccharide (LPS), revealed a previously unreported, transient rise in intracellular low molecular weight (LMW)·Fe59 complex ([LMW·Fe]i) at ≤2 min returning to the basal level within 15 min. The [LMW·Fe]i response preceded IκB kinase (IKK) (≥15 min) and NF-κB (≥30 min) activation. Iron chelators (1,2-dimethyl-3-hydroxy-pyridin-4-one and N,N′-bis-2-hydroxybenzylethylenediamine-N,N′-diacetic acid) abrogated the [LMW·Fe]i response and IKK and NF-κB activation. The [LMW·Fe]i response was also observed in tumor necrosis factor α (TNFα)-stimulated HMs and RAW264.7 cells treated with LPS and interferon-γ but not in primary rat hepatocytes or myofibroblastic cells exposed to LPS or TNFα. Both [LMW·Fe]i response and IKK activation in LPS-stimulated HMs were inhibited by diphenylene iodonium (nonspecific inhibitor for flavin-containing oxidases), L-N6(1-iminoethyl)lysine (selective iNOS inhibitor), and adenoviral-mediated expression of a dominant negative mutant of Rac1 or Cu,Zn-superoxide dismutase, suggesting the role of .NO and O2 .- in mediating the iron signaling. In fact, this inhibition was recapitulated by a cell-permeable scavenger of ONOO-, 5,10,15,20-tetrakis (4-sulfonatophenyl)porphyrinato iron (III) chloride. Conversely, ONOO- alone induced both [LMW·Fe]i response and IKK activation. Finally, direct addition of ferrous iron to cultured HMs activated IKK and NF-κB. These results support a novel signaling role for [LMW·Fe]i in IKK activation, which appears to be induced by ONOO- and selectively operative in macrophages.

Original languageEnglish (US)
Pages (from-to)17646-17654
Number of pages9
JournalJournal of Biological Chemistry
Volume278
Issue number20
DOIs
StatePublished - May 16 2003
Externally publishedYes

    Fingerprint

ASJC Scopus subject areas

  • Biochemistry

Cite this

Xiong, S., She, H., Takeuchi, H., Han, B., Engelhardt, J. F., Barton, C. H., Zandi, E., Giulivi, C. R., & Tsukamoto, H. (2003). Signaling role of intracellular iron in NF-κb activation. Journal of Biological Chemistry, 278(20), 17646-17654. https://doi.org/10.1074/jbc.M210905200