title: [Effect of hypoxia on the activation and visfatin expression in rat hepatic stellate cells].
abstract: OBJECTIVE
  To investigate the effect of hypoxia on the visfatin and the expression of smooth muscle-actin (alpha-SMA) and hypoxia-inducible factor-1alpha (HIF-1alpha) in rat hepatic stellate cells (HSCs).
  
  
  METHODS
  Rat primary HSCs 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. Completely activated primary HSCs were exposed to hypoxic conditions (37 degrees C, 5% CO2, 1% O2, 94% N2), or normoxic conditions (37 degrees C, 5% CO2, 21% O2, 74% N2), for 3, 6, 12 or 24 h respectively. The expression of alpha-SMA, the marker of HSC activation, and visfatin were assessed by Real time-PCR and Western blot. The Expression of HIF-1alpha was detected by Real time-PCR.
  
  
  RESULTS
  HIF-1alpha mRNA in rat HSCs was induced after exposed to hypoxia for 3 h, and maintained elevated status up to 24 h. HSCs exposed to 1% O2 hypoxic conditions for 6 h increased alpha-SMA mRNA and protein expression. Visfatin mRNA expression was up-regulated after subjected to hypoxia for 12 h, and protein level was elevated after 6 h hypoxia. A positive linear correlation existed between alpha-SMA and visfatin expression in responsible to hypoxia (r = 0.991 (genes) and r = 0.968 (proteins), P < 0.05).
  
  
  CONCLUSIONS
  Microcirculation impairment could significantly induce alpha-SMA and visfatin expression in rat HSCs, which might potentate the activation process of HSCs.
authors.forename: Zhi-Hui
authors.initials: ZH
authors.lastname: Yi
authors.forename: Ruo-Ting
authors.initials: RT
authors.lastname: Men
authors.forename: Cong
authors.initials: C
authors.lastname: Yuan
authors.forename: Yong-Jun
authors.initials: YJ
authors.lastname: Zhu
authors.forename: Mao-Yao
authors.initials: MY
authors.lastname: Wen
authors.forename: Li
authors.initials: L
authors.lastname: Yang
mesh_terms: D000199:Actins / Q000378:metabolism*; D000818:Animals; D015687:Cell Hypoxia; D002478:Cells, Cultured; D055166:Hepatic Stellate Cells / Q000166:cytology* / Q000378:metabolism; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism; D054409:Nicotinamide Phosphoribosyltransferase / Q000378:metabolism*; D012333:RNA, Messenger; D051381:Rats; D017207:Rats, Sprague-Dawley; D015854:Up-Regulation
chemical_list: D000199:Actins; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D054409:Nicotinamide Phosphoribosyltransferase; D012333:RNA, Messenger; C497444:Hif1a protein, rat; C498862:Acta2 protein, rat