Mitochondrial uncouplers inhibit hepatic stellate cell activation

Eduardo L Guimarães, Jan Best, Laurent Dollé, Mustapha Najimi, Etienne Sokal, Leo A van Grunsven

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

BACKGROUND: Mitochondrial dysfunction participates in the progression of several pathologies. Although there is increasing evidence for a mitochondrial role in liver disease, little is known about its contribution to hepatic stellate cell (HSC) activation. In this study we investigated the role of mitochondrial activity through mild uncoupling during in vitro activation of HSCs.

METHODS: Cultured primary human and mouse HSCs were treated with the chemical uncouplers FCCP and Valinomycin. ATP levels were measured by luciferase assay and production of reactive oxygen species was determined using the fluorescent probe DCFH-DA. Possible cytotoxicity by uncoupler treatment was evaluated by caspase 3/7 activity and cytoplasmic protease leakage. Activation of HSCs and their response to the pro-fibrogenic cytokine TGF-β was evaluated by gene expression of activation markers and signal mediators using RT-qPCR. Proliferation was measured by incorporation of EdU and protein expression of α-smooth muscle actin was analyzed by immunocytochemistry and western blot.

RESULTS: FCCP and Valinomycin treatment mildly decreased ATP and reactive oxygen species levels. Both uncouplers increased the expression of mitochondrial genes such as Tfam and COXIV while inducing morphological features of quiescent mouse HSCs and abrogating TGF-β signal transduction. Mild uncoupling reduced HSC proliferation and expression of pro-fibrogenic markers of mouse and human HSCs.

CONCLUSIONS: Mild mitochondrial uncoupling inhibits culture-induced HSC activation and their response to pro-fibrogenic cytokines like TGF-β. These results therefore suggest mitochondrial uncoupling of HSCs as a strategy to reduce progression of liver fibrosis.

Original languageEnglish
Article number68
Number of pages12
JournalBMC Gastroenterology
Volume12
DOIs
Publication statusPublished - 11 Jun 2012

Keywords

  • Actins
  • Adenosine Triphosphate
  • Animals
  • Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
  • Caspase 3
  • Caspase 7
  • Cell Proliferation
  • Cell Survival
  • Cells, Cultured
  • Hepatic Stellate Cells
  • Humans
  • In Vitro Techniques
  • Male
  • Mice
  • Mitochondria, Liver
  • Models, Animal
  • Reactive Oxygen Species
  • Uncoupling Agents
  • Valinomycin
  • Journal Article
  • Research Support, Non-U.S. Gov't

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