Iron primes 3T3-L1 adipocytes to a TLR4-mediated inflammatory response

Nutrition. 2015 Oct;31(10):1266-74. doi: 10.1016/j.nut.2015.04.007. Epub 2015 May 5.

Abstract

Objective: Iron participates in several mechanisms involving inflammation and innate immunity, yet the dysregulation of its homeostasis is a major cause of metabolic syndrome. Adipocytes should play a major role in iron metabolism, as an impairment in iron turnover is closely related to insulin resistance, obesity, and type 2 diabetes. The aim of this study was to investigate the role of iron in an in vitro-inflamed adipocyte model.

Methods: Gene expression of tumor necrosis factor-α, interleukin-6, inflammatory chemokines (CCL3, CCL4, and CXCL12), and molecules involved in iron metabolism were evaluated in an in vitro mouse 3T3-L1 cell model. Cells underwent treatment with FeSO4 heptahydrate and lipopolysaccharide (LPS) stimulation. Toll-like receptor 4 (TLR4) membrane expression, lipid droplet immunohystochemistry, and lipolysis were also evaluated.

Results: Iron sulphate heptahydrate elicited gene expression of hepcidin, hemojuvelin, and ferroportin at different time courses. Additionally, it activated lipolysis but did not trigger any adipokine gene expression. When cells treated with physiological doses of iron were also stimulated with LPS, an enhancement in the LPS-induced gene expression of cytokines and chemokines was observed. The enhancement occurred with different patterns depending on different time courses and investigated genes, showing its maximal effect for IL-6 gene expression.

Conclusions: FeSO4 heptahydrate at a relatively physiological dose, induced gene expression of iron modulatory proteins and also enhanced RNA transcripts of several inflammatory cytokines and chemokines through a priming/synergistic mechanism involving membrane TLR4.

Keywords: Adipocyte: Lipopolysaccharide; Chemokine; Inflammation; Iron; Toll-like receptors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3-L1 Cells / metabolism*
  • Adipocytes / immunology*
  • Animals
  • Cation Transport Proteins / metabolism
  • Chemokines / metabolism
  • Cytokines / metabolism
  • Diabetes Mellitus, Type 2 / metabolism
  • Ferric Compounds / administration & dosage
  • Ferric Compounds / metabolism
  • GPI-Linked Proteins
  • Gene Expression*
  • Hemochromatosis Protein
  • Hepcidins / metabolism
  • Inflammation / metabolism
  • Insulin Resistance
  • Iron / administration & dosage
  • Iron / metabolism*
  • Lipopolysaccharides / administration & dosage
  • Lipopolysaccharides / metabolism
  • Membrane Proteins / metabolism
  • Mice
  • Obesity / metabolism
  • Toll-Like Receptor 4 / immunology*

Substances

  • Cation Transport Proteins
  • Chemokines
  • Cytokines
  • Ferric Compounds
  • GPI-Linked Proteins
  • HJV protein, mouse
  • Hemochromatosis Protein
  • Hepcidins
  • Lipopolysaccharides
  • Membrane Proteins
  • Tlr4 protein, mouse
  • Toll-Like Receptor 4
  • metal transporting protein 1
  • ferric sulfate
  • Iron