title: Differential effects of Smad3 targeting in a murine model of chronic kidney disease.
abstract: Transforming growth factor (TGF)-β1 has a pivotal role in the pathogenesis of progressive kidney diseases that are characterized by fibrosis. The main intracellular signaling pathway of TGF-β1 is the Smad system, where Smad2 and Smad3 play a central role in transcriptional regulation of target genes involved in extracellular matrix (ECM) metabolism. This study analyzes the hypothesis that blockade of Smad3 attenuates the development of TGF-β1-driven renal fibrosis. This was examined in vivo in a transgenic model of TGF-β1-induced chronic kidney disease with Smad3 or without Smad3 expression and in vitro in mesangial cells and glomerular endothelial cells with Smad2/3 inhibitors or Smad3-knockdown. Electron microscopy was used for evaluation of morphological changes, real-time polymerase chain reaction for detection of RNA expression, and immunohistochemistry for localization of ECM components. Matrix metalloproteinase (MMP) level was assessed by gelatin zymography electrophoresis and located by in situ zymography. The results show TGF-β1-induced mesangial matrix expansion, tubulointerstitial fibrosis, and tubular basement membrane thickening that are attenuated by Smad3 deletion, whereas TGF-β1-induced glomerular basement membrane thickening is not shown. The amount and distribution profile of MMP-2 may suggest a role of the enzyme herein. We conclude that Smad3 targeting is not exclusively beneficial as Smad3 has diverse transcriptional regulatory effects in different cell types in the kidney.
authors.affiliation: Research Laboratory for Biochemical Pathology, Aarhus University Hospital, Institute of Clinical Medicine, University of Aarhus, Aarhus, Denmark.
authors.forename: Terese
authors.initials: T
authors.lastname: Kellenberger
authors.affiliation: Research Laboratory for Biochemical Pathology, Aarhus University Hospital, Institute of Clinical Medicine, University of Aarhus, Aarhus, Denmark.
authors.forename: Søren
authors.initials: S
authors.lastname: Krag
authors.affiliation: Department of Connective Tissue Biology, Institute of Biomedicine, University of Aarhus, Aarhus, Denmark.
authors.forename: Carl Christian
authors.initials: CC
authors.lastname: Danielsen
authors.affiliation: Pharmacology and Cancer Biology, Duke University Medical Center, Durham, NC.
authors.forename: Xiao-Fan
authors.initials: XF
authors.lastname: Wang
authors.affiliation: Stereology and Electron Microscopy Laboratory, Centre for Stochastic Geometry and Advanced Bioimaging, Institute of Clinical Medicine, University of Aarhus, Aarhus, Denmark.
authors.forename: Jens Randel
authors.initials: JR
authors.lastname: Nyengaard
authors.affiliation: Research Laboratory for Biochemical Pathology, Aarhus University Hospital, Institute of Clinical Medicine, University of Aarhus, Aarhus, Denmark.
authors.forename: Lea
authors.initials: L
authors.lastname: Pedersen
authors.affiliation: Department of Molecular Biology, University of Aarhus, Aarhus, Denmark.
authors.forename: Chuanxu
authors.initials: C
authors.lastname: Yang
authors.affiliation: Department of Molecular Biology, University of Aarhus, Aarhus, Denmark.
authors.forename: Shan
authors.initials: S
authors.lastname: Gao
authors.affiliation: Research Laboratory for Biochemical Pathology, Aarhus University Hospital, Institute of Clinical Medicine, University of Aarhus, Aarhus, Denmark.
authors.forename: Lise
authors.initials: L
authors.lastname: Wogensen
keywords: Extracellular matrix; fibrosis; matrix metalloproteinase; transforming growth factor‐β1
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