Hepcidin knockout mice fed with iron-rich diet develop chronic liver injury and liver fibrosis due to lysosomal iron overload

J Hepatol. 2014 Sep;61(3):633-41. doi: 10.1016/j.jhep.2014.04.034. Epub 2014 May 6.

Abstract

Background & aims: Hepcidin is the central regulator of iron homeostasis and altered hepcidin signalling results in both hereditary and acquired iron overload. While the association between iron overload and development of end-stage liver disease is well established, the underlying mechanisms are largely unknown. To improve that, we analysed hepcidin knockout (KO) mice as a model of iron overload-associated liver disease.

Methods: Hepcidin wild type (WT) and KO mice fed with 3% carbonyl iron-containing diet starting at one month of age were compared to age-matched animals kept on standard chow. Liver histology and serum parameters were used to assess the extent of liver injury and fibrosis. Iron distribution was determined by subcellular fractionation and electron microscopy.

Results: Among mice kept on iron-rich diet, 6 months old hepcidin KO mice (vs. WT) displayed profound hepatic iron overload (3,186 ± 411 vs. 1,045 ± 159 μg/mg tissue, p<0.005), elevated liver enzymes (ALT: KO 128 ± 6, WT 56 ± 5 IU/L, p<0.05), mild hepatic inflammation and hepatocellular apoptosis. Twelve, but not six months old KO mice fed with iron-rich diet developed moderate liver fibrosis. The liver injury was accompanied by a marked lysosomal iron overload and lysosomal fragility with release of cathepsin B into the cytoplasm. Increased p62 levels and autofluorescent iron complexes suggested impaired protein degradation. As a mechanism leading to lysosomal iron overload, the autophagy (lysosomal influx) was increased.

Conclusions: Hepcidin KO mice represent a novel model of iron overload-related liver diseases and implicate lysosomal injury as a crucial event in iron toxicity.

Keywords: Autophagy; Hemochromatosis; Inflammation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / physiology
  • Disease Models, Animal
  • Hepatic Stellate Cells / pathology
  • Hepatic Stellate Cells / physiology
  • Hepcidins / deficiency*
  • Hepcidins / genetics
  • Hepcidins / physiology
  • Homeostasis / physiology
  • Iron / metabolism*
  • Iron, Dietary / adverse effects*
  • Liver / enzymology
  • Liver / pathology
  • Liver Cirrhosis / etiology*
  • Liver Cirrhosis / metabolism
  • Liver Cirrhosis / physiopathology
  • Lung Injury / etiology*
  • Lung Injury / metabolism
  • Lung Injury / physiopathology
  • Lysosomes / metabolism*
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Time Factors

Substances

  • Hepcidins
  • Iron, Dietary
  • Iron