title: [Effect of p65 gene inhibited by siRNA on differention of rat marrow mesenchymal stem cells into neurons].
abstract: OBJECTIVE
  To investigate the effect of p65 gene inhibited by siRNA on neuronic differentiation in the marrow mesenchymal stem cells (MSCs).
  
  
  METHODS
  The MSCs were transfected with Rn-p65-siRNA. Fasudil hydrochloride induced MSCs differentiating into neurons. The non-transfected group and negative control group (transfected with negative control siRNA marked by Cy3) were used as controls. The fluorescence expressed by transfected MSCs were observed under inverted fluorescence microscope at 24 h,48 h and 72 h after transfected with negative control siRNA. The viability of MSCs was detected by MTT at 24 h, 48 h and 72 h after transfected with Rn-p65-siRNA. The expressions of p65 mRNA and protein in MSCs were detected by RT-PCR and Western blot respectively. The expressions of p65 protein, NSE, MAP-2 and glial fibrillary acidic protein (GFAP) were detected by immunocytochemical method after transfection for 6 h.
  
  
  RESULTS
  The fluorescence of MSCs was mostly displayed after transfection of 72 hours and the efficiency of transfection was up to 83.3% ± 3.8%. Meanwhile, the p65 mRNA and p65 protein expressed by MSCs of transfected group were significantly decreased (P < 0.05); MTT displayed that the viability of MSCs was also significantly reduced (P < 0.05). The best efficiency of induction was observed in the transfected group. There were higher expressions of NSE and MAP-2 than the other group (P < 0.05).
  
  
  CONCLUSIONS
  The p65 gene inhibited by siRNA can promote the marrow mesenchymal stem cells to differentiate into neurons.
authors.forename: Er-yi
authors.initials: EY
authors.lastname: Zhao
authors.forename: Yan-jie
authors.initials: YJ
authors.lastname: Jia
authors.forename: Dai-mei
authors.initials: DM
authors.lastname: Wang
authors.forename: Guo-qiang
authors.initials: GQ
authors.lastname: Wen
authors.forename: Wen-juan
authors.initials: WJ
authors.lastname: Guan
authors.forename: Li-jun
authors.initials: LJ
authors.lastname: Jing
authors.forename: Yi-dong
authors.initials: YD
authors.lastname: Deng
mesh_terms: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives; D000818:Animals; D002454:Cell Differentiation; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D009474:Neurons / Q000166:cytology*; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D051381:Rats; D051996:Transcription Factor RelA / Q000037:antagonists & inhibitors* / Q000378:metabolism; D014162:Transfection
chemical_list: D005904:Glial Fibrillary Acidic Protein; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C498145:Rela protein, rat; D051996:Transcription Factor RelA; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; C049347:fasudil