title: Influence of Expression Plasmid of Connective Tissue Growth Factor and Tissue Inhibitor of Metalloproteinase-1 shRNA on Hepatic Precancerous Fibrosis in Rats.
abstract: BACKGROUND
  In this study, influence caused by expression plasmids of connective tissue growth factor (CTGF) and tissue inhibitor of metalloproteinase-1 (TIMP-1) short hairpin RNA (shRNA) on mRNA expression of CTGF,TIMP-1,procol-α1 and PCIII in hepatic tissue with hepatic fibrosis, a precancerous condition, in rats is analyzed.
  
  
  METHODS
  To screen and construct shRNA expression plasimid which effectively interferes RNA targets of CTGF and TIMP-1 in rats. 50 cleaning Wistar male rats are allocated randomly at 5 different groups after precancerous fibrosis models and then injection of shRNA expression plasimids. Plasmid psiRNA-GFP-Com (CTGF and TIMP-1 included), psiRNA-GFP-CTGF, psiRNA-GFP-TIMP-1 and psiRNA- DUO-GFPzeo of blank plasmid are injected at group A, B, C and D, respectively, and as model control group that none plasimid is injected at group E. In 2 weeks after last injection, to hepatic tissue at different groups, protein expression of CTGF, TIMP-1, procol-α1and PC III is tested by immunohistochemical method and,mRNA expression of CTGF,TIMP-1,procol-α1 and PCIII is measured by real-time PCR. One-way ANOVA is used to comparison between-groups.
  
  
  RESULTS
  Compared with model group, there is no obvious difference of mRNA expression among CTGF,TIMP-1,procol-α1,PC III and of protein expression among CTGF, TIMP-1, procol-α1, PC III in hepatic tissue at group injected with blank plasmid. Expression quantity of mRNA of CTGF, TIMP-1, procol-α1 and PCIII at group A, B and C decreases, protein expression of CTGF, TIMP-1, procol-α1, PC III in hepatic tissue is lower, where the inhibition of combination RNA interference group (group A) on procol-α1 mRNA transcription and procol-α1 protein expression is superior to that of single interference group (group B and C) (P<0.01 or P<0.05).
  
  
  CONCLUSIONS
  RNA interference on CTGF and/or TIMP-1 is obviously a inhibiting factor for mRNA and protein expression of CTGF, TIMP-1, procol-α1 and PCIII. Combination RNA interference on genes of CTGF and TIMP-1 is superior to that of single RNA interference, and this could be a contribution for prevention of precancerous condition.
authors.affiliation: Division of Infectious Diseases, Affiliated Zhongda Hospital of Southeast University, Nanjing, Jiangsu Province, China E-mail : slim888@163.com; huangxinen06@163.com.
authors.forename: Qun
authors.initials: Q
authors.lastname: Zhang
authors.forename: Fu-Li
authors.initials: FL
authors.lastname: Shu
authors.forename: Yu-Feng
authors.initials: YF
authors.lastname: Jiang
authors.forename: Xin-En
authors.initials: XE
authors.lastname: Huang
mesh_terms: D000818:Animals; D055513:Connective Tissue Growth Factor / Q000037:antagonists & inhibitors* / Q000235:genetics; D008103:Liver Cirrhosis / Q000235:genetics / Q000473:pathology / Q000517:prevention & control*; D008297:Male; D010957:Plasmids / Q000008:administration & dosage*; D011230:Precancerous Conditions / Q000235:genetics / Q000473:pathology / Q000517:prevention & control*; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D051381:Rats; D017208:Rats, Wistar; D060888:Real-Time Polymerase Chain Reaction; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000037:antagonists & inhibitors* / Q000235:genetics
chemical_list: D012333:RNA, Messenger; D034741:RNA, Small Interfering; D019715:Tissue Inhibitor of Metalloproteinase-1; D055513:Connective Tissue Growth Factor