title: [Construction of shRNA expressing plasmid targeted rat NogoB and observation of the effection about NogoB on hepatic stellate cells].
abstract: OBJECTIVE
  To construct shRNA expressing plasmid inhibiting rat NogoB and to observe its possible effect on rat primary hepatic stellate cells (HSCs) contraction.
  
  
  METHODS
  Three pairs of shRNAs targeting different sequence of rat NogoB were designed and constructed into pSuper plasmid by DNA recombination technique. Culture-activated HSCs were transfected with NogoB-shRNA plasmids to scan the effective plasmid which could inhibit NogoB gene expression by Real-time PCR. And this depressant effect was also confirmed with Western blot. After NogoB was knocked-down effectively, ETA and ETB mRNA expression were assessed by Real-time PCR.
  
  
  RESULTS
  Among the three pairs of recombinant plasmids, NogoB-shRNA2 plasmid could inhibit NogoB expression specifically. In HSCs, NogoB knockdown decreased the ratio of ETA and ETB.
  
  
  CONCLUSIONS
  We constructed specific NogoB-shRNA expression plasmid successfully which might be involved in contraction of HSCs.
authors.forename: Mao-yao
authors.initials: MY
authors.lastname: Wen
authors.forename: Ruo-ting
authors.initials: RT
authors.lastname: Men
authors.forename: Xue-lian
authors.initials: XL
authors.lastname: Dan
authors.forename: Wen-chao
authors.initials: WC
authors.lastname: Wu
authors.forename: Li
authors.initials: L
authors.lastname: Yang
mesh_terms: D000818:Animals; D015870:Gene Expression; D055166:Hepatic Stellate Cells / Q000166:cytology*; D009185:Myelin Proteins / Q000235:genetics*; D000070798:Nogo Proteins; D010957:Plasmids; D034622:RNA Interference; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D051381:Rats; D014162:Transfection
chemical_list: D009185:Myelin Proteins; D000070798:Nogo Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C000603197:Rtn4 protein, rat