title: Hyperglycemia-Induced Modulation of the Physiognomy and Angiogenic Potential of Fibroblasts Mediated by Matrix Metalloproteinase-2: Implications for Venous Stenosis Formation Associated with Hemodialysis Vascular Access in Diabetic Milieu.
abstract: OBJECTIVE
  It is hypothesized that venous stenosis formation associated with hemodialysis vascular-access failure is caused by hypoxia-mediated fibroblast-to-myofibroblast differentiation accompanied by proliferation and migration, and that diabetic patients have worse clinical outcomes. The aim of this study was to determine the functional and gene expression outcomes of matrix metalloproteinase-2 (Mmp-2) silencing in fibroblasts cultured under hyperglycemia and euglycemia with hypoxic and normoxic stimuli.
  
  
  METHODS
  AKR-2B fibroblasts were stably transduced using lentivirus-mediated shRNA-Mmp-2 or scrambled controls and subjected to hypoxia or normoxia under hyperglycemic or euglycemic conditions for 24 and 72 h. Gene expression of vascular endothelial growth factor-A (Vegf-A), Vegfr-1, Mmp-2, Mmp-9 and tissue inhibitors of matrix metalloproteinases (Timps) were determined by RT-PCR. Collagen I and IV secretion and cellular proliferation and migration were determined.
  
  
  RESULTS
  Under hyperglycemic conditions, there is a significant reduction in the average gene expression of Vegf-A and Mmp-9, with an increase in Timp-1 at 24 h of hypoxia (p < 0.05) in Mmp-2-silenced fibroblasts when compared to controls. In addition, there is a decrease in collagen I and IV secretion and cellular migration. The euglycemic cells were able to reverse these findings.
  
  
  CONCLUSIONS
  These findings demonstrate the rationale for using anti-Mmp-2 therapy in dialysis patients with hemodialysis vascular access in helping to reduce stenosis formation.
authors.affiliation: Amity Institute of Public Health, Amity University Uttar Pradesh, Noida, Uttar Pradesh, India.
authors.forename: Rajiv
authors.initials: R
authors.lastname: Janardhanan
authors.forename: Sreenivasulu
authors.initials: S
authors.lastname: Kilari
authors.forename: Edward B
authors.initials: EB
authors.lastname: Leof
authors.forename: Sanjay
authors.initials: S
authors.lastname: Misra
mesh_terms: D000818:Animals; D001166:Arteriovenous Shunt, Surgical / Q000009:adverse effects*; D015687:Cell Hypoxia; D002460:Cell Line; D002465:Cell Movement; D049109:Cell Proliferation; D024042:Collagen Type I / Q000378:metabolism; D024141:Collagen Type IV / Q000378:metabolism; D017077:Culture Media, Conditioned / Q000378:metabolism; D003925:Diabetic Angiopathies / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D005347:Fibroblasts / Q000201:enzymology* / Q000473:pathology; D005786:Gene Expression Regulation; D005947:Glucose / Q000378:metabolism*; D006083:Graft Occlusion, Vascular / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism; D051379:Mice; D058628:Myofibroblasts / Q000201:enzymology / Q000473:pathology; D009389:Neovascularization, Pathologic; D034622:RNA Interference; D006435:Renal Dialysis; D015398:Signal Transduction; D013997:Time Factors; D019714:Tissue Inhibitor of Metalloproteinases / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000235:genetics / Q000378:metabolism
chemical_list: D024042:Collagen Type I; D024141:Collagen Type IV; D017077:Culture Media, Conditioned; D019714:Tissue Inhibitor of Metalloproteinases; D042461:Vascular Endothelial Growth Factor A; C467480:vascular endothelial growth factor A, mouse; C501163:Flt1 protein, mouse; D040281:Vascular Endothelial Growth Factor Receptor-1; D020778:Matrix Metalloproteinase 2; C522362:Mmp2 protein, mouse; D020780:Matrix Metalloproteinase 9; C501830:Mmp9 protein, mouse; D005947:Glucose