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1 Department of Nephrology, Goteborg University, Gothenburg, Sweden
2 Wallenberg laboratory for Cardiovascular Research, Matrix and Proteoglycan, Goteborg University, Gothenburg, Sweden
* To whom correspondence should be addressed. E-mail: anna.bjornson{at}kidney.med.gu.se.
This article describes the possible role of the endothelial cell surface coat, containing proteoglycans (PGs) with connected glycosaminoglycans (GAGs), in maintaining glomerular permselectivity. Primary human glomerular endothelial cells (HGEC) in culture were treated with the nephrosis-inducing agent puromycin aminonucleoside (PAN). Analysis was made by Taqman® real-time PCR, western blotting and by metabolic labeling with [35S]-sulfate. The HGECs express several PGs: syndecan, versican, glypican, perlecan, decorin and biglycan which may contribute to the glomerular charge barrier. PAN treatment down-regulated both the protein expression (by 25%) and the mRNA expression (by 37 ±6 % (p<0.001, n=8)) of versican in comparison to control. Transferases important for chondroitin and heparan sulfate biosynthesis were also significantly down-regulated by PAN, resulting in less sulfate groups, shorter GAG chains and reduced PG net negative charge. Moreover, analysis of the cell media after PAN treatment revealed a reduced content of [35S]-sulfate labeled PGs (40% of control). We conclude that PAN may cause proteinuria by affecting the endothelial cell surface layer and not only by disrupting the foot process arrangement of the podocytes. Thus, the endothelium may be a more important component of the glomerular barrier than hitherto acknowledged.
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