title: [Role of macrophage migration inhibitory factor in hepatic stellate cells activation].
abstract: OBJECTIVE
  To investigate the role of macrophage migration inhibitory factor (MIF) in activation of primary hepatic stellate cells (HSC), and to explore the blocking effect of specific antisense oligonucleotides (ASONs) targeting MIF on the activation of primary HSCs.
  
  
  METHODS
  Rat primary HSC were isolated from SD rats by in situ perfusion of collagenase and pronase and single-step Nycodenz density gradient centrifugation, and then cultured and activated. The expression of alpha-smooth muscle actin (alpha-SMA), the marker of HSC activation, was assessed by RT-PCR and immunocytochemistry (ICC). The expression of MIF was detected by RT-PCR, enzyme-linked immuno sorbent assay (ELISA), immunofluorescence (IF) or ICC. Completely activated primary HSC were transfected with MIF-specific antisense oligonucleotide (MIF-ASONs), mis-sense oligonucleotides of MIF, or blank liposome (negative-control).
  
  
  RESULTS
  Quiescent HSC expressed very low level of MIF, while MIF gene and protein expression increased significantly during the process of HSC activation. Positive correlation was found between the expression of alpha-SMA and MIF (r = 0.944, P < 0.01). Compared with the control, activated primary Gen HSC transfected with MIF-ASONs showed lower MIF and alpha-SMA expressions at gene and protein profiles (P < 0.05).
  
  
  CONCLUSIONS
  The expression of MIF was up-regulated in the process of HSC activation. Activation in culture of primarily isolated rat HSC could be inhibited by MIF-ASONs, suggesting at least in part that, MIF might be a novel and important factor involved in HSC activation and hepatic fibrosis.
authors.affiliation: Department of Gastroenterology, West China Hospital, Sichuan University, Chengdu 610041, China.
authors.forename: Yong-Jun
authors.initials: YJ
authors.lastname: Zhu
authors.forename: Xiao-Lin
authors.initials: XL
authors.lastname: Hu
authors.forename: Ruo-Ting
authors.initials: RT
authors.lastname: Men
authors.forename: Li
authors.initials: L
authors.lastname: Yang
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D055166:Hepatic Stellate Cells / Q000166:cytology* / Q000378:metabolism*; D019746:Intramolecular Oxidoreductases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D008103:Liver Cirrhosis / Q000503:physiopathology; D008263:Macrophage Migration-Inhibitory Factors / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D008297:Male; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D061251:Primary Cell Culture; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D014162:Transfection; D015854:Up-Regulation
chemical_list: D000199:Actins; D019746:Intramolecular Oxidoreductases; D008263:Macrophage Migration-Inhibitory Factors; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; C498862:Acta2 protein, rat; C515599:Mif protein, rat