title: A role for autophagy during hepatic stellate cell activation.
abstract: OBJECTIVE
  Autophagy is a metabolic process that degrades and recycles intracellular organelles and proteins with many connections to human disease and physiology. We studied the role of autophagy during hepatic stellate cell (HSC) activation, a key event in liver fibrogenesis.
  
  
  METHODS
  Analysis of the autophagic flux during in vitro activation of primary mouse HSCs was performed using a DsRed-GFP-LC3B encoding plasmid. The effect of autophagy inhibition by bafilomycin A1 on the in vitro activation process of human and mouse HSCs was examined by measuring proliferation, presence of activation markers by RT-qPCR, immunofluorescence, and Western blotting. Analysis of lipid droplet and microtubule-associated protein light chain 3 beta (LC3B) colocalization in the presence of PDGF-BB was investigated by immunocytochemistry.
  
  
  RESULTS
  A significant increased autophagic flux was observed during culture induced mouse HSC activation. Treatment of mouse HSCs and human HSCs with autophagy inhibitor bafilomycin A1 results in a significant decreased proliferation and expression of activation markers. In addition, lipid droplets and LC3B colocalization was increased after PDGF-BB treatment in quiescent HSCs.
  
  
  CONCLUSIONS
  During HSC activation, autophagic flux is increased. The demonstration of partly inhibition of in vitro HSC activation after treatment with an autophagy inhibitor unveils a potential new therapeutic strategy for liver fibrosis.
authors.affiliation: Department of Cell Biology, Vrije Universiteit Brussel, Laarbeeklaan 103, Brussels 1090, Belgium.
authors.forename: Lien F R
authors.initials: LF
authors.lastname: Thoen
authors.forename: Eduardo L M
authors.initials: EL
authors.lastname: Guimarães
authors.forename: Laurent
authors.initials: L
authors.lastname: Dollé
authors.forename: Inge
authors.initials: I
authors.lastname: Mannaerts
authors.forename: Mustapha
authors.initials: M
authors.lastname: Najimi
authors.forename: Etienne
authors.initials: E
authors.lastname: Sokal
authors.forename: Leo A
authors.initials: LA
authors.lastname: van Grunsven
mesh_terms: D000199:Actins / Q000235:genetics; D000818:Animals; D001343:Autophagy / Q000187:drug effects / Q000502:physiology*; D000077214:Becaplermin; D002251:Carbon Tetrachloride / Q000633:toxicity; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D024042:Collagen Type I / Q000235:genetics; D000090602:Collagen Type I, alpha 1 Chain; D055166:Hepatic Stellate Cells / Q000187:drug effects / Q000473:pathology / Q000502:physiology*; D006801:Humans; D066298:In Vitro Techniques; D050356:Lipid Metabolism / Q000187:drug effects; D008103:Liver Cirrhosis / Q000209:etiology / Q000473:pathology / Q000503:physiopathology; D018942:Macrolides / Q000494:pharmacology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008249:Protein-Lysine 6-Oxidase; D020574:Proto-Oncogene Proteins c-sis / Q000378:metabolism / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: C541116:ACTA2 protein, human; D000199:Actins; D024042:Collagen Type I; D000090602:Collagen Type I, alpha 1 Chain; D018942:Macrolides; C480860:Map1lc3b protein, mouse; D008869:Microtubule-Associated Proteins; D020574:Proto-Oncogene Proteins c-sis; D012333:RNA, Messenger; D000077214:Becaplermin; C040929:bafilomycin A1; D002251:Carbon Tetrachloride; D008249:Protein-Lysine 6-Oxidase