title: Heparin increasing podocyte-specific gene expressions.
abstract: OBJECTIVE
  Heparin, a highly sulfated glycosaminoglycan, has been shown to have a renoprotective effect on renal diseases, but its mechanisms remain to be elucidated. In this study, we examined the effect of heparin on podocytes by using primary cultured podocytes positive for podocyte-specific markers including podocin and podocalyxin.
  
  
  METHODS
  Podocytes were cultured from highly purified glomeruli isolated by the method with renal perfusion with magnetic beads and digestion of collagenase. Podocyte-specific gene expressions and proteins were examined by real-time polymerase chain reaction (PCR), Western blotting and immunofluorescence microscopy.
  
  
  RESULTS
  Real-time PCR showed that addition of heparin to the culture media significantly upregulated most of the podocyte-specific genes in a dose-dependent and time-dependent manner. Western blotting showed a marked increase in protein levels of nephrin and podocin. Podocin localization at cell-cell contact sites became conspicuous in the presence of heparin. The effect of heparin was observed even in culture media deprived of bovine foetal serum. Heparan sulfate, less sulfated than heparin, and hyaluronan did not show such effects, but sulfated dextran did markedly.
  
  
  CONCLUSIONS
  Heparin acts on cultured podocytes to increase podocyte-specific gene expressions. A high degree of sulfation is crucial for the effect of heparin.
authors.affiliation: Department of Structural Pathology, Institute of Nephrology, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan.
authors.forename: Eishin
authors.initials: E
authors.lastname: Yaoita
authors.forename: Yutaka
authors.initials: Y
authors.lastname: Yoshida
authors.forename: Masaaki
authors.initials: M
authors.lastname: Nameta
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.forename: Hidehiko
authors.initials: H
authors.lastname: Fujinaka
authors.forename: Sameh
authors.initials: S
authors.lastname: Magdeldin
authors.forename: Bo
authors.initials: B
authors.lastname: Xu
authors.forename: Tadashi
authors.initials: T
authors.lastname: Yamamoto
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D016264:Dextran Sulfate / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation / Q000187:drug effects*; D006493:Heparin / Q000494:pharmacology*; D006497:Heparan Sulfate / Q000494:pharmacology; D006820:Hyaluronic Acid / Q000494:pharmacology; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D050199:Podocytes / Q000187:drug effects* / Q000378:metabolism; D061251:Primary Cell Culture; D012333:RNA, Messenger / Q000378:metabolism; D017208:Rats, Wistar; D013997:Time Factors; D015854:Up-Regulation
chemical_list: D016264:Dextran Sulfate; D006493:Heparin; D006497:Heparan Sulfate; D006820:Hyaluronic Acid; D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; D012333:RNA, Messenger; C407223:NPHS2 protein; C113267:nephrin
keywords: culture; heparin; nephrin; podocyte; sulfation