title: Silibinin Inhibits Proliferation and Migration of Human Hepatic Stellate LX-2 Cells.
abstract: BACKGROUND
  Proliferation of hepatic stellate cells (HSCs) play pivotal role in the progression of hepatic fibrosis consequent to chronic liver injury. Silibinin (SBN), a flavonoid compound, has shown to possess cell cycle arresting potential against many actively proliferating cancers cell lines. The objective of this study was to evaluate the anti-proliferative and cell cycle arresting properties of SBN in rapidly proliferating human hepatic stellate LX-2 cell line.
  
  
  METHODS
  LX-2 cells were fed with culture medium supplemented with different concentrations of SBN (10, 50 and 100 μM). After 24 and 96 h of treatment, total cell number was determined by counting. Cytotoxicity was evaluated by trypan blue dye exclusion test. The expression profile of cMyc and peroxisome proliferator-activated receptor-γ (PPAR-γ) protein expressions was evaluated by Western blotting. Oxidative stress marker genes profile was quantified using qPCR. The migratory response of HSCs was observed by scrape wound healing assay.
  
  
  RESULTS
  SBN treatments significantly inhibit the LX-2 cell proliferation (without affecting its viability) in dose dependent manner. This treatment also retards the migration of LX-2 cells toward injured area. In Western blotting studies SBN treatment up regulated the protein expressions of PPAR-γ and inhibited cMyc.
  
  
  CONCLUSIONS
  The present study shows that SBN retards the proliferation, activation and migration of LX-2 cells without inducing cytotoxicity and oxidative stress. The profound effects could be due to cell cycle arresting potential of SBN.
authors.affiliation: Institut de Recherche Expérimentale et Clinique (IREC), Laboratory of Pediatric Hepatology and Cell Therapy, Université Catholique de Louvain, 1200 Brussels, Belgium.
authors.forename: Devaraj
authors.initials: D
authors.lastname: Ezhilarasan
authors.affiliation: Institut de Recherche Expérimentale et Clinique (IREC), Laboratory of Pediatric Hepatology and Cell Therapy, Université Catholique de Louvain, 1200 Brussels, Belgium.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Evraerts
authors.affiliation: Louvain Drug Research Institute, Toxicology and Cancer Biology Research Group, PMNT Unit, Université Catholique de Louvain, 1200 Brussels, Belgium.
authors.forename: Sid
authors.initials: S
authors.lastname: Brice
authors.affiliation: Facultad de Ciencias de la Salud, Universidad Arturo Prat, Iquique, Chile.
authors.forename: Pedro
authors.initials: P
authors.lastname: Buc-Calderon
authors.affiliation: Department of Pharmacology and Environmental Toxicology, Food and Hepatotoxicology Laboratory, Dr. ALM Post Graduate Institute of Basic Medical Sciences, University of Madras, Taramani Campus, Chennai 600 113, India.
authors.forename: Sivanesan
authors.initials: S
authors.lastname: Karthikeyan
authors.affiliation: Institut de Recherche Expérimentale et Clinique (IREC), Laboratory of Pediatric Hepatology and Cell Therapy, Université Catholique de Louvain, 1200 Brussels, Belgium.
authors.forename: Etienne
authors.initials: E
authors.lastname: Sokal
authors.affiliation: Institut de Recherche Expérimentale et Clinique (IREC), Laboratory of Pediatric Hepatology and Cell Therapy, Université Catholique de Louvain, 1200 Brussels, Belgium.
authors.forename: Mustapha
authors.initials: M
authors.lastname: Najimi
keywords: AKR1C1, aldo-keto reductase family 1, member C1; ARE, antioxidant responsive element; CDKI, cyclin dependent kinase inhibitor; CYP450, cytochrome P450; DMEM, Dulbecco's modified Eagle's medium; DMSO, dimethylsulphoxide; ECM, extracellular matrix; FBS, fetal bovine serum; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; HMOX1, heme oxygenase (decycling) 1; HSCs, hepatic stellate cells; NQO1, NAD(P)H dehydrogenase, quinone 1; Nrf-2, nuclear respiratory factor; PPAR-γ, peroxisome proliferator-activated receptor-γ; PPIA, peptidylprolyl isomerase A; ROS, reactive oxygen species; SBN, silibinin; TXNRD1, thioredoxin reductase 1; cytotoxicity; hepatic stellate cells; oxidative stress; qPCR, quantitative polymerase chain reaction; wound healing