P311 modulates hepatic stellate cells migration

Liver Int. 2015 Apr;35(4):1253-64. doi: 10.1111/liv.12691. Epub 2014 Oct 10.

Abstract

Background & aims: Liver fibrosis is induced by the accumulation of extracellular matrix, deposited mainly by activated hepatic stellate cells (HSCs). One key characteristic of stellate cell activation is the directional migration to the site of injury during the wound-healing process. P311 is a protein that has been shown to play a role in migration and we aimed to study a possible role for this protein during stellate cell migration.

Methods: Mouse stellate cells were isolated and cultured in vitro to investigate P311 protein and gene expression during HSC activation by immunocytochemistry and RT-qPCR respectively. Expression of P311 during in vivo activation was evaluated in CCl4 and bile duct ligation-induced liver fibrosis. Production of reactive oxygen species was determined using the fluorescent probe DCFH-DA. By siRNA-mediated knockdown of P311, we investigated a possible effect on proliferation by incorporation of EdU and on migration by Boyden chamber assays.

Results: P311 gene expression was increased during both in vitro and in vivo activation of HSCs. siRNA-mediated knockdown led to a decrease in reactive oxygen production and cell proliferation. Migration induced by different chemokines, such as PDGF-bb and MCP-1 was inhibited by knockdown of P311.

Conclusions: P311 is central to reactive oxygen species-mediated HSC migration induced by different chemokines.

Keywords: chemokines; liver fibrosis; migration.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Becaplermin
  • Cell Movement* / drug effects
  • Cell Proliferation
  • Cells, Cultured
  • Chemokine CCL2 / pharmacology
  • Gene Expression Regulation
  • Heme Oxygenase-1 / genetics
  • Heme Oxygenase-1 / metabolism
  • Hepatic Stellate Cells / drug effects
  • Hepatic Stellate Cells / metabolism*
  • Hepatic Stellate Cells / pathology
  • Liver / drug effects
  • Liver / metabolism*
  • Liver / pathology
  • Liver Cirrhosis, Experimental / genetics
  • Liver Cirrhosis, Experimental / metabolism*
  • Liver Cirrhosis, Experimental / pathology
  • Male
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Mice, Inbred BALB C
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / metabolism*
  • Oxidative Stress
  • Proto-Oncogene Proteins c-sis / pharmacology
  • RNA Interference
  • Reactive Oxygen Species / metabolism
  • Signal Transduction
  • Time Factors
  • Transfection
  • Transforming Growth Factor beta / pharmacology

Substances

  • Chemokine CCL2
  • Membrane Proteins
  • Nerve Tissue Proteins
  • P311 protein, mouse
  • Proto-Oncogene Proteins c-sis
  • Reactive Oxygen Species
  • Transforming Growth Factor beta
  • Becaplermin
  • Heme Oxygenase-1
  • Hmox1 protein, mouse