title: Attenuation of fibrosis in vitro and in vivo with SPARC siRNA.
abstract: BACKGROUND
  SPARC is a matricellular protein, which, along with other extracellular matrix components including collagens, is commonly over-expressed in fibrotic diseases. The purpose of this study was to examine whether inhibition of SPARC can regulate collagen expression in vitro and in vivo, and subsequently attenuate fibrotic stimulation by bleomycin in mouse skin and lungs.
  
  
  METHODS
  In in vitro studies, skin fibroblasts obtained from a Tgfbr1 knock-in mouse (TBR1CA; Cre-ER) were transfected with SPARC siRNA. Gene and protein expressions of the Col1a2 and the Ctgf were examined by real-time RT-PCR and Western blotting, respectively. In in vivo studies, C57BL/6 mice were induced for skin and lung fibrosis by bleomycin and followed by SPARC siRNA treatment through subcutaneous injection and intratracheal instillation, respectively. The pathological changes of skin and lungs were assessed by hematoxylin and eosin and Masson's trichrome stains. The expression changes of collagen in the tissues were assessed by real-time RT-PCR and non-crosslinked fibrillar collagen content assays.
  
  
  RESULTS
  SPARC siRNA significantly reduced gene and protein expression of collagen type 1 in fibroblasts obtained from the TBR1CA; Cre-ER mouse that was induced for constitutively active TGF-beta receptor I. Skin and lung fibrosis induced by bleomycin was markedly reduced by treatment with SPARC siRNA. The anti-fibrotic effect of SPARC siRNA in vivo was accompanied by an inhibition of Ctgf expression in these same tissues.
  
  
  CONCLUSIONS
  Specific inhibition of SPARC effectively reduced fibrotic changes in vitro and in vivo. SPARC inhibition may represent a potential therapeutic approach to fibrotic diseases.
authors.affiliation: State Key Laboratory of Genetic Engineering and MOE Key Laboratory of Contemporary Anthropology, School of Life Sciences, Fudan University, 220 Handan Road, Shanghai 200433, PR China.
authors.forename: Jiu-Cun
authors.initials: JC
authors.lastname: Wang
authors.forename: Syeling
authors.initials: S
authors.lastname: Lai
authors.forename: Xinjian
authors.initials: X
authors.lastname: Guo
authors.forename: Xuefeng
authors.initials: X
authors.lastname: Zhang
authors.forename: Benoit
authors.initials: B
authors.lastname: de Crombrugghe
authors.forename: Sonali
authors.initials: S
authors.lastname: Sonnylal
authors.forename: Frank C
authors.initials: FC
authors.lastname: Arnett
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Zhou
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D055513:Connective Tissue Growth Factor / Q000235:genetics / Q000378:metabolism; D004195:Disease Models, Animal; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D005355:Fibrosis; D015870:Gene Expression; D055879:Gene Knock-In Techniques; D020868:Gene Silencing; D008168:Lung / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D015676:Osteonectin / Q000037:antagonists & inhibitors / Q000235:genetics*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics; D011658:Pulmonary Fibrosis / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D034741:RNA, Small Interfering / Q000235:genetics* / Q000494:pharmacology; D000077293:Receptor, Transforming Growth Factor-beta Type I; D018125:Receptors, Transforming Growth Factor beta / Q000235:genetics; D012867:Skin / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D014162:Transfection
chemical_list: C526318:CCN2 protein, mouse; D024042:Collagen Type I; D015676:Osteonectin; D034741:RNA, Small Interfering; D018125:Receptors, Transforming Growth Factor beta; D055513:Connective Tissue Growth Factor; D017346:Protein Serine-Threonine Kinases; D000077293:Receptor, Transforming Growth Factor-beta Type I; C000625842:Tgfbr1 protein, mouse