title: Relaxin-expressing adenovirus decreases collagen synthesis and up-regulates matrix metalloproteinase expression in keloid fibroblasts:  in vitro experiments.
abstract: BACKGROUND
  The hormone relaxin has been shown to affect the extracellular matrix by inhibiting collagen synthesis and expression in fibroblasts stimulated with a profibrotic agent. It also increases matrix metalloproteinase (MMP) expression. To investigate its effect on expression of collagen and MMPs in keloid fibroblasts and human dermal fibroblasts, the authors introduced a relaxin-expressing adenovirus (dE1-RGD/lacZ/RLX) into a human dermal fibroblast cell line and keloid fibroblasts.
  
  
  METHODS
  Both fibroblasts were infected with dE1-RGD/lacZ/RLX or control virus, and protein levels of relaxin and secreted transforming growth factor (TGF)-β1 were assessed by enzyme-linked immunosorbent assay, and mRNA levels of collagen type I, collagen type III, MMP-1, and MMP-3 were assessed by real-time reverse-transcriptase polymerase chain reaction and enzyme-linked immunosorbent assay. Expression of Smad3 and phosphorylated Smad3 was also examined, and relaxin's effect on Smad2/3 complex localization was evaluated.
  
  
  RESULTS
  When human dermal fibroblasts and keloid fibroblasts were transduced with dE1-RGD/lacZ/RLX or dE1-RGD/lacZ (control), mRNA expression of type I and type III collagen was markedly decreased by relaxin regardless of TGF-β (10 ng/ml) treatment. Expression of Smad3 and phosphorylated Smad3 was reduced in keloid fibroblasts and decreased translocation of Smad 2/3 complex from cytosols to the nucleus of the human dermal fibroblasts with TGF-β after dE1-RGD/lacZ/RLX transduction, suggesting that relaxin reduces collagen synthesis by blocking TGF-β signaling. Analyses revealed that MMP-1 and MMP-3 expression were significantly increased in human dermal fibroblasts and keloid fibroblasts after dE1-RGD/lacZ/RLX transduction.
  
  
  CONCLUSIONS
  These results suggest that the antifibrotic effect of relaxin-expressing adenovirus may have therapeutic effects on keloids.
authors.affiliation: Seoul, Korea From the Institute for Human Tissue Restoration, Department of Plastic and Reconstructive Surgery, and the Department of Dermatology, Yonsei University College of Medicine; and the Department of Bioengineering, College of Engineering, Hanyang University.
authors.forename: Won Jai
authors.initials: WJ
authors.lastname: Lee
authors.forename: Il-Kyu
authors.initials: IK
authors.lastname: Choi
authors.forename: Ju Hee
authors.initials: JH
authors.lastname: Lee
authors.forename: Jung-Sun
authors.initials: JS
authors.lastname: Lee
authors.forename: Yong Oock
authors.initials: YO
authors.lastname: Kim
authors.forename: Dong Kyun
authors.initials: DK
authors.lastname: Rah
authors.forename: Chae-Ok
authors.initials: CO
authors.lastname: Yun
mesh_terms: D000256:Adenoviridae / Q000378:metabolism*; D002460:Cell Line; D002478:Cells, Cultured; D003094:Collagen / Q000096:biosynthesis*; D020405:Dermis / Q000166:cytology / Q000378:metabolism / Q000821:virology; D005109:Extracellular Matrix / Q000378:metabolism*; D005347:Fibroblasts / Q000378:metabolism / Q000821:virology*; D006801:Humans; D007627:Keloid / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D045726:Metalloproteases / Q000378:metabolism*; D012065:Relaxin / Q000378:metabolism*; D051900:Smad3 Protein / Q000378:metabolism; D015854:Up-Regulation
chemical_list: C497790:SMAD3 protein, human; D051900:Smad3 Protein; D012065:Relaxin; D003094:Collagen; D045726:Metalloproteases