title: Selective modulation of the secretion of proteinases and their inhibitors by growth factors in cultured differentiated podocytes.
abstract: Selective modulation of the secretion of proteinases and their inhibitors by growth factors in cultured differentiated podocytes.
  
  
  BACKGROUND
  Podocyte damage is considered to be an important factor in the development of glomerulosclerosis. Morphological studies on experimental models of progressive glomerular disease have identified the detachment of podocytes from the glomerular basement membrane (GBM) as a critical step in the development and progression of glomerulosclerosis. Degradation of the GBM by proteinases also might be a potential mechanism of the detachment because the process impairs the connection between podocytes and the GBM. The present study examined the effects of basic fibroblast growth factor (bFGF), transforming growth factor-beta1 (TGF-beta1) and platelet-derived growth factor (PDGF) on the secretion of proteinases [cathepsin L and matrix metalloproteinases (MMPs)] and their inhibitors [cystatin C and tissue inhibitor of metalloproteinase-2 (TIMP-2)] from differentiated podocytes in culture.
  
  
  METHODS
  Expression of mRNAs for receptors of growth factors (bFGF, PDGF, TGF-beta1), the proteinases and their inhibitors in differentiated podocytes were shown by RT-PCR. The secretion of cathepsin L, cystatin C and TIMP-2 from differentiated podocytes were shown by immunoblot analysis. The activities of MMPs-2 and -9 from differentiated podocytes were shown by gelatin zymography.
  
  
  RESULTS
  Expression of mRNAs for receptors of the growth factors, the proteinases and their inhibitors were confirmed. bFGF increased the secretion of cathepsin L (5.04-fold at 20 ng/mL), but did not alter the secretion of its extracellular inhibitor, cystatin C. In contrast, TGF-beta1 increased the activities of MMPs-2 and -9 (3.23-fold at 10 ng/mL and 25.3-fold at 10 ng/mL, respectively) from differentiated podocytes, but did not enhance the secretion of its inhibitor, TIMP-2. In addition, bFGF enhanced the secretion of TIMP-2 (2.75-fold at 20 ng/mL) and TGF-beta1 enhanced the secretion of cystatin C (2.32-fold at 20 ng/mL). These results demonstrate the imbalance of the secretion of proteinases and their inhibitors after incubation of such growth factors. Of particular interest was the observation of differences in regulation of proteinases and their extracellular inhibitors in response to bFGF and TGF-beta1. PDGF only slightly increased the secretion of cathepsin L (2.54-fold at 20 ng/mL) but exerted no effect on the secretion of cystatin C, MMPs, and TIMP-2 from differentiated podocytes.
  
  
  CONCLUSIONS
  These results indicate, to our knowledge for the first time, that in differentiated podocytes, both cathepsin L and its inhibitor are independently regulated by different growth factors. It appears that increases in proteolytic activities may induce degradation of the glomerular basement membrane (GBM), which plays an important role in the progression of glomerulosclerosis.
authors.affiliation: Division of Nephrology, Department of Internal Medicine, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo 113-8421, Japan.
authors.forename: Katsuhiko
authors.initials: K
authors.lastname: Asanuma
authors.forename: Isao
authors.initials: I
authors.lastname: Shirato
authors.forename: Kazumi
authors.initials: K
authors.lastname: Ishidoh
authors.forename: Eiki
authors.initials: E
authors.lastname: Kominami
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Tomino
mesh_terms: D000818:Animals; D056668:Cathepsin L; D002403:Cathepsins / Q000378:metabolism; D002454:Cell Differentiation; D002461:Cell Line, Transformed; D055316:Cystatin C; D015891:Cystatins / Q000378:metabolism; D003546:Cysteine Endopeptidases; D005110:Extracellular Space / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D015870:Gene Expression; D006133:Growth Substances / Q000494:pharmacology*; D007678:Kidney Glomerulus / Q000166:cytology / Q000378:metabolism*; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D051379:Mice; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D017978:Receptors, Growth Factor / Q000235:genetics; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000378:metabolism*; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D053773:Transforming Growth Factor beta1
chemical_list: C525031:Cst3 protein, mouse; D055316:Cystatin C; D015891:Cystatins; D006133:Growth Substances; D010982:Platelet-Derived Growth Factor; D012333:RNA, Messenger; D017978:Receptors, Growth Factor; C508961:Tgfb1 protein, mouse; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D016222:Fibroblast Growth Factor 2; D019716:Tissue Inhibitor of Metalloproteinase-2; D002403:Cathepsins; D003546:Cysteine Endopeptidases; D056668:Cathepsin L; C534278:Ctsl protein, mouse; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9