title: Elastase, but not proteinase 3 (PR3), induces proteinuria associated with loss of glomerular basement membrane heparan sulphate after in vivo renal perfusion in rats.
abstract: Elastase, but not PR3, induces proteinuria associated with loss of glomerular basement membrane (GBM) heparan sulphate after in vivo renal perfusion in rats. PR3 and elastase are cationic neutral serine proteinases present in the azurophilic granules of polymorphonuclear leucocytes. Release of these proteolytic enzymes along the glomerular capillary wall may induce glomerular injury. Here, we investigated the effects of PR3 and elastase on the induction of proteinuria and glomerular injury after renal perfusion of these enzymes in Brown-Norway rats. Perfusion of active elastase induced a dose-dependent proteinuria 24h after perfusion, while inactivated elastase did not. Perfusion of comparable amounts of active PR3 did not induce proteinuria. Light and electron microscopy showed no morphological abnormalities in any experimental group. However, immunohistology revealed that proteinuria occurring after perfusion of active elastase was associated with a strong reduction in intraglomerular expression of the heparan sulphate side chain and, to a lesser extent, of the protein core of heparan sulphate proteoglycans (HSPG). In vitro, both elastase and PR3 digested HSPG. However, PR3 bound to a lesser extent to HSPG than elastase. We conclude that elastase, but not PR3, induces proteinuria after in vivo renal perfusion. This differential effect probably relates to different binding to the GBM of those enzymes due to differences in their isoelectric points. Degradation of heparan sulfate proteoglycans, leading to the disappearance of their side chains that contribute to the polyanionic structure of the GBM, appears to be involved in the induction of proteinuria after perfusion of elastase.
authors.affiliation: Department of Clinical Immunology, University Hospital, Groningen, The Netherlands.
authors.forename: P
authors.initials: P
authors.lastname: Heeringa
authors.forename: J
authors.initials: J
authors.lastname: Van den Born
authors.forename: E
authors.initials: E
authors.lastname: Brouwer
authors.forename: K M
authors.initials: KM
authors.lastname: Dolman
authors.forename: P A
authors.initials: PA
authors.lastname: Klok
authors.forename: M G
authors.initials: MG
authors.lastname: Huitema
authors.forename: P C
authors.initials: PC
authors.lastname: Limburg
authors.forename: M A
authors.initials: MA
authors.lastname: Bakker
authors.forename: J H
authors.initials: JH
authors.lastname: Berden
authors.forename: M R
authors.initials: MR
authors.lastname: Daha
authors.forename: C G
authors.initials: CG
authors.lastname: Kallenberg
mesh_terms: D000818:Animals; D001485:Basement Membrane / Q000187:drug effects / Q000378:metabolism; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate / Q000378:metabolism*; D007678:Kidney Glomerulus / Q000187:drug effects* / Q000378:metabolism; D053146:Myeloblastin; D010196:Pancreatic Elastase / Q000494:pharmacology*; D010477:Perfusion; D011507:Proteinuria / Q000139:chemically induced*; D011509:Proteoglycans / Q000378:metabolism*; D051381:Rats; D011914:Rats, Inbred BN; D012697:Serine Endopeptidases / Q000494:pharmacology*
chemical_list: D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate; D053146:Myeloblastin; D010196:Pancreatic Elastase; D011509:Proteoglycans; D012697:Serine Endopeptidases