title: Role of connective growth factor in plasminogen activator inhibitor-1 and fibronectin expression induced by transforming growth factor beta1 in renal tubular cells.
abstract: BACKGROUND
  Connective tissue growth factor (CTGF) contributes greatly to renal tubulointerstitial fibrosis, which is the final event leading to end-stage renal failure. This study was designed to investigate the effects of CTGF antisense oligodeoxynucleotides (ODNs) on the expressions of plasminogen activator inhibitor-1 (PAI-1) and fibronectin in renal tubular cells induced by transforming growth factor beta1 (TGF-beta1) in addition to the role of CTGF in the accumulation and degradation of renal extracellular matrix (ECM).
  
  
  METHODS
  A human proximal tubular epithelial cell line (HKC) was cultured in vitro. Cationic lipid-mediated CTGF antisense ODNs were transfected into HKC cells. After HKC cells were stimulated with TGF-beta1 (5 microg/L), the mRNA levels of PAI-1 and fibronectin were measured by RT-PCR. Intracellular PAI-1 protein synthesis was assessed by flow cytometry. The secreted PAI-1 and fibronectin in the medium were determined by Western blot and ELISA, respectively.
  
  
  RESULTS
  TGF-beta1 was found to induce tubular CTGF, PAI-1, and fibronectin mRNA expression. PAI-1 and fibronectin mRNA expression induced by TGF-beta1 was significantly inhibited by CTGF antisense ODNs. CTGF antisense ODNs also inhibited intracellular PAI-1 protein synthesis and lowered the levels of PAI-1 and fibronectin protein secreted into the medium.
  
  
  CONCLUSIONS
  CTGF may play a crucial role in the accumulation and degradation of excessive ECM during tubulointerstitial fibrosis, and transfecting CTGF antisense ODNs may be an effective way to prevent renal fibrosis.
authors.affiliation: Department of Nephrology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China. zhangchun1234@hotmail.com
authors.forename: Chun
authors.initials: C
authors.lastname: Zhang
authors.forename: Xian-fang
authors.initials: XF
authors.lastname: Meng
authors.forename: Zhong-hua
authors.initials: ZH
authors.lastname: Zhu
authors.forename: Xiao
authors.initials: X
authors.lastname: Yang
authors.forename: An-guo
authors.initials: AG
authors.lastname: Deng
mesh_terms: D002478:Cells, Cultured; D055513:Connective Tissue Growth Factor; D005109:Extracellular Matrix / Q000378:metabolism; D005353:Fibronectins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics / Q000502:physiology*; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000502:physiology*; D007684:Kidney Tubules / Q000378:metabolism*; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D017395:Plasminogen Activator Inhibitor 1 / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D053773:Transforming Growth Factor beta1
chemical_list: C526317:CCN2 protein, human; D005353:Fibronectins; D017874:Immediate-Early Proteins; D036341:Intercellular Signaling Peptides and Proteins; D016376:Oligonucleotides, Antisense; D017395:Plasminogen Activator Inhibitor 1; D012333:RNA, Messenger; C508960:TGFB1 protein, human; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D055513:Connective Tissue Growth Factor