title: Matrix metalloproteinase 9 induces endothelial-mesenchymal transition via Notch activation in human kidney glomerular endothelial cells.
abstract: Endothelial-mesenchymal transition (EndoMT) is a major source of myofibroblast formation in kidney fibrosis. Our previous study showed a profibrotic role for matrix metalloproteinase 9 (MMP-9) in kidney fibrosis via induction of epithelial-mesenchymal transition (EMT). Inhibition of MMP-9 activity reduced kidney fibrosis in murine unilateral ureteral obstruction. This study investigated whether MMP-9 also plays a role in EndoMT in human glomerular endothelial cells.
  
  
  
  TGF-β1 (10 or 20 ng/ml) induced EndoMT in HKGECs as shown by morphological changes. In addition, VE-cadherin and CD31 were significantly downregulated, whereas α-SMA, vimentin, and N-cadherin were upregulated. RT-PCR revealed that Snail, a known inducer of EMT, was upregulated. The MMP inhibitor GM6001 abrogated TGF-β1-induced EndoMT. Zymography indicated that MMP-9 was also upregulated in TGF-β1-treated HKGECs. Recombinant MMP-9 (2 μg/ml) induced EndoMT in HKGECs via Notch signaling, as evidenced by increased formation of the Notch intracellular domain (NICD) and decreased Notch 1. Inhibition of MMP-9 activity by its inhibitor showed a dose-dependent response in preventing TGF-β1-induced α-SMA and NICD in HKGECs, whereas inhibition of Notch signaling by γ-secretase inhibitor (GSI) blocked rMMP-9-induced EndoMT.
  
  
  
  Taken together, our results demonstrate that MMP-9 plays an important role in TGF-β1-induced EndoMT via upregulation of Notch signaling in HKGECs.
authors.affiliation: Centre for Transplant and Renal Research, Westmead Institute for Medical Research, the University of Sydney, 176 Hawkesbury Road, Sydney, NSW, 2145, Australia.|The School of Biomedical Sciences, Chengdu Medical College, Chengdu, 610500, PR China.
authors.forename: Ye
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Centre for Transplant and Renal Research, Westmead Institute for Medical Research, the University of Sydney, 176 Hawkesbury Road, Sydney, NSW, 2145, Australia.|Department of Nephrology, Second Hospital of Shanxi Medical University, Shanxi Kidney Disease Institute, WuYi Road 382, Taiyuan, 030001, Shanxi, PR China.
authors.forename: Xi
authors.initials: X
authors.lastname: Qiao
authors.affiliation: Department of Nephrology, Second Hospital of Shanxi Medical University, Shanxi Kidney Disease Institute, WuYi Road 382, Taiyuan, 030001, Shanxi, PR China.
authors.forename: Lihua
authors.initials: L
authors.lastname: Wang
authors.affiliation: Centre for Transplant and Renal Research, Westmead Institute for Medical Research, the University of Sydney, 176 Hawkesbury Road, Sydney, NSW, 2145, Australia.
authors.forename: Tian Kui
authors.initials: TK
authors.lastname: Tan
authors.affiliation: Department of Biochemistry and Molecular Biology, Shanxi Medical University, Xinjian Road 56, Taiyuan, 030001, Shanxi, PR China.
authors.forename: Hong
authors.initials: H
authors.lastname: Zhao
authors.affiliation: Experimental Centre of Science and Research, the First Clinical Hospital of Shanxi Medical University, Xinjian Road 382, Taiyuan, 030001, Shanxi, PR China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Biochemistry and Molecular Biology, Shanxi Medical University, Xinjian Road 56, Taiyuan, 030001, Shanxi, PR China.
authors.forename: Jianlin
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Centre for Transplant and Renal Research, Westmead Institute for Medical Research, the University of Sydney, 176 Hawkesbury Road, Sydney, NSW, 2145, Australia.
authors.forename: Padmashree
authors.initials: P
authors.lastname: Rao
authors.affiliation: Centre for Transplant and Renal Research, Westmead Institute for Medical Research, the University of Sydney, 176 Hawkesbury Road, Sydney, NSW, 2145, Australia.
authors.forename: Qi
authors.initials: Q
authors.lastname: Cao
authors.affiliation: Centre for Transplant and Renal Research, Westmead Institute for Medical Research, the University of Sydney, 176 Hawkesbury Road, Sydney, NSW, 2145, Australia.
authors.forename: Yiping
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Centre for Transplant and Renal Research, Westmead Institute for Medical Research, the University of Sydney, 176 Hawkesbury Road, Sydney, NSW, 2145, Australia.
authors.forename: Ya
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Centre for Kidney Research, Children's Hospital at Westmead, 212 Hawkesbury Road, Sydney, NSW, Australia.
authors.forename: Yuan Min
authors.initials: YM
authors.lastname: Wang
authors.affiliation: Centre for Transplant and Renal Research, Westmead Institute for Medical Research, the University of Sydney, 176 Hawkesbury Road, Sydney, NSW, 2145, Australia.
authors.forename: Vincent W S
authors.initials: VW
authors.lastname: Lee
authors.affiliation: Centre for Kidney Research, Children's Hospital at Westmead, 212 Hawkesbury Road, Sydney, NSW, Australia.
authors.forename: Stephen I
authors.initials: SI
authors.lastname: Alexander
authors.affiliation: Centre for Transplant and Renal Research, Westmead Institute for Medical Research, the University of Sydney, 176 Hawkesbury Road, Sydney, NSW, 2145, Australia.
authors.forename: David C H
authors.initials: DC
authors.lastname: Harris
authors.affiliation: Centre for Transplant and Renal Research, Westmead Institute for Medical Research, the University of Sydney, 176 Hawkesbury Road, Sydney, NSW, 2145, Australia. guoping.zheng@sydney.edu.au.
authors.forename: Guoping
authors.initials: G
authors.lastname: Zheng
mesh_terms: D053829:Amyloid Precursor Protein Secretases / Q000037:antagonists & inhibitors / Q000378:metabolism; D004151:Dipeptides / Q000494:pharmacology; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007678:Kidney Glomerulus / Q000166:cytology*; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D061965:Matrix Metalloproteinase Inhibitors / Q000494:pharmacology; D008648:Mesoderm / Q000166:cytology* / Q000187:drug effects; D051880:Receptors, Notch / Q000378:metabolism*; D011994:Recombinant Proteins / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology
chemical_list: D004151:Dipeptides; D061965:Matrix Metalloproteinase Inhibitors; C078131:N-(2(R)-2-(hydroxamidocarbonylmethyl)-4-methylpentanoyl)-L-tryptophan methylamide; D051880:Receptors, Notch; D011994:Recombinant Proteins; D053773:Transforming Growth Factor beta1; D053829:Amyloid Precursor Protein Secretases; D020780:Matrix Metalloproteinase 9
keywords: Endothelial-mesenchymal transition; Human glomerular endothelial cells; Matrix metalloproteinase 9; Notch; TGF-β1