title: [Effect of RNA interference on CTGF expression and collagen synthesis induced by hypoxia in rat cardiac fibroblasts].
abstract: OBJECTIVE
  To investigate the effect of the lentiviral vector pGCL-GFP transferred connective tissue growth factor (CTGF) short hairpin RNA (CTGF-ShRNA) on CTGF expression, cell proliferation and collagen synthesis induced by hypoxia in rat cardiac fibroblasts (CFs).
  
  
  METHODS
  CTGF-ShRNA plasmids successfully constructed and screened. CFs of adult Sprague-Dawley (SD) rats isolated with the method of trypsin digestion and differential anchoring velocity which randomly divided into the control group, the hypoxia group, Hypo+pGCL-GFP group and Hypo+CTGF-ShRNA group. The mRNA and protein levels of CTGF were detected by means of semi-quantitative reverse transcriptional polymerase chain reaction (RT-PCR) and Western blot 24 h later. Proliferation of CFs was observed by WST-1 coloricmetric assay and synthesis of collagen was observed by the hydroxyproline.
  
  
  RESULTS
  Successfully constructed and screened CTGF short hairpin RNA. Compared with control group, the expression of CTGF mRNA and protein levels induced by hypoxia in CFs were markedly up-regulated in Hypoxia, Hypo + pGCL-GFP and Hypo + CTGF-ShRNA group (P < 0.05). CFs proliferation and collagen synthesis in Hypoxia, Hypo+pGCL-GFP and Hypo+CTGF-ShRNA group were significantly higher than that of the control group (P < 0.05). In comparison to Hypoxia and Hypo+pGCL-GFP group, the CTGF mRNA and protein levels induced by hypoxia in CFs were markedly down-regulated in Hypo + CTGF-ShRNA group (P < 0.01). CFs proliferation and collagen synthesis in Hypo+CTGF-ShRNA group were significantly lower than that of the Hypoxia and Hypo+pGCL-GFP group (P < 0.01).
  
  
  CONCLUSIONS
  CTGF mRNA and protein expression, CFs proliferation and collagen synthesis induced by hypoxia in CFs effectively inhibited by CTGF-ShRNA.
authors.affiliation: Institutes of GCP, West China Hospital, Sichuan University, Chengdu 610041, China.
authors.forename: Jia-yang
authors.initials: JY
authors.lastname: Sun
authors.forename: Chun-tao
authors.initials: CT
authors.lastname: Liu
authors.forename: Ni-wen
authors.initials: NW
authors.lastname: Huang
mesh_terms: D000818:Animals; D015687:Cell Hypoxia; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis*; D055513:Connective Tissue Growth Factor / Q000235:genetics / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism*; D005355:Fibrosis / Q000209:etiology; D016086:Lentivirus / Q000235:genetics / Q000378:metabolism; D008297:Male; D009206:Myocardium / Q000166:cytology* / Q000378:metabolism / Q000473:pathology; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: C526316:CCN2 protein, rat; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D055513:Connective Tissue Growth Factor; D003094:Collagen