Laboratory Investigation

Kidney International (1996) 49, 1079–1089; doi:10.1038/ki.1996.157

Effects of high glucose on the production of heparan sulfate proteoglycan by mesangial and epithelial cells

Nicole F van Det, Jacob van den Born, Jouke T Tamsma, Nicole A M Verhagen, Jo H M Berden, Jan A Bruijn, Mohamed R Daha and Fokko J van der Woude

Departments of Nephrology, Endocrinology, and Pathology, University Hospital Leiden, Leiden, and Department of Nephrology, University Hospital, Nijmegen, Nijmegen, The Netherlands

Correspondence: Nicole van Det, University Hospital Leiden, Department of Nephrology, Building 1, C3P, P.O. Box 9600, 2300 RC Leiden, The Netherlands.

Received 19 June 1995; Revised 1 November 1995; Accepted 2 November 1995.

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Abstract

Effects of high glucose on the production of heparan sulfate proteoglycan by mesangial and epithelial cells. Changes in heparan sulfate metabolism may be important in the pathogenesis of diabetic nephropathy. Recent studies performed on renal biopsies from patients with diabetic nephropathy revealed a decrease in heparan sulfate glycosaminoglycan staining in the glomerular basement membrane without changes in staining for heparan sulfate proteoglycan-core protein. To understand this phenomenon at the cellular level, we investigated the effect of high glucose conditions on the synthesis of heparan sulfate proteoglycan by glomerular cells in vitro. Human adult mesangial and glomerular visceral epithelial cells were cultured under normal (5 mM) and high glucose (25 mM) conditions. Immunofluorescence performed on cells cultured in 25 mM glucose confirmed and extended the in vivo histological observations. Using metabolic labeling we observed an altered proteoglycan production under high glucose conditions, with predominantly a decrease in heparan sulfate compared to dermatan sulfate or chondroitin sulfate proteoglycan. N-sulfation analysis of heparan sulfate proteoglycan produced under high glucose conditions revealed less di- and tetrasaccharides compared to larger oligosaccharides, indicating an altered sulfation pattern. Furthermore, with quantification of glomerular basement membrane heparan sulfate by ELISA, a significant decrease was observed when mesangial and visceral epithelial cells were cultured in high glucose conditions. We conclude that high glucose concentration induces a significant alteration of heparan sulfate production by mesangial cells and visceral epithelial cells. Changes in sulfation and changes in absolute quantities are both observed and may explain the earlier in vivo observations. These changes may be of importance for the altered integrity of the glomerular charge-dependent filtration barrier and growth-factor matrix interactions in diabetic nephropathy.

Abbreviations:

HS, heparan sulfate; HSPG, heparan sulfate proteoglycan; GAG, glycosaminoglycan; DN, diabetic nephropathy; MC, mesangial cells; GVEC, glomerular visceral epithelial cells; DMEM, Dulbecco's modified Eagle's medium

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