Article

Increased expression of heparan sulfate proteoglycan on the cultured renal epithelial cells during oxalate exposure.

Department of Urology, Kurume University School of Medicine, Japan.
The Kurume Medical Journal 01/2008; 55(1-2):19-28. DOI: 10.2739/kurumemedj.55.19
Source: PubMed

ABSTRACT We have previously reported that heparan sulfate (HS) / heparan sulfate proteoglycan (HSPG, syndecan-1) expression significantly increased in the rat kidney during calcium oxalate (CaOx) nephrolithiasis. Although the exact mechanism of the increased syndecan expression still remains unclear, HS/syndecan is thought to have some important roles in CaOx crystal formation. The present study examined the role of HS during oxalate exposure by using a newly developed cell line (KIC-synd-1) that expresses human heparan sulfate proteoglycan (syndecan-1). Quantitative competitive (QC)-RT-PCR was used to examine change of syndecan-1 mRNA expression in KIC-synd-1 cells. Production of syndecan-1 core protein and glycosaminoglycans (GAGs) were also confirmed by Western blot, immunohistochemistry and HPLC, respectively. Wild type Mardin-Darby canine kidney (MDCK) cells were also examined in the same manner. The stable expression of syndecan-1 gene and production of both core protein and HS chains were confirmed in the newly developed KIC-synd-1 cell line. Increased syndecan-1 mRNA expression and production of core proteins were confirmed in KIC-synd-1 cells during oxalate exposure. MTT assay revealed that the cell viability decreased significantly in the MDCK cells after 1 mM oxalate exposure (p<0.05). On the other hand, there was no significant difference in the oxalate exposed KIC-synd-1 cells. However, the cell viability in KIC-synd-1 cells pretreated with heparitinase digestion decreased significantly before oxalate exposure (p<0.05). The present data suggests that both exogenous and endogenous HS exerts protective effect against oxalate-induced cell injuries. Previous studies in our laboratory have indicated that hyperoxaluria and deposition of CaOx crystals resulted in renal tubular cellular injury inducing the synthesis of HSPG to protect and repair the damaged epithelial cell surface. The present data offers strong support for this hypothesis. Finally, HS could be potent inhibitor of CaOx nephrolithiasis and the absence of this substance on the tubular surface may increase the risk of CaOx crystal formation and retention.

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