Background: Neuropilin-1 (NRP1) is a transmembrane glycoprotein, initially defined as a receptor for members of the semaphorin family. We observed that NRP1 expression was downregulated by the addition of advanced glycation end products-modified bovine serum albumin (AGE-BSA). The present study was undertaken to unravel the molecular mechanisms underlying AGE-BSA-mediated NRP1 suppression. Methods: Expression of NRP1 was analyzed in podocytes. The transcriptional activity of the NRP1 promoter was investigated using wild-type and mutant NRP1 promoter reporter constructs. Electrophoretic mobility assays were performed. Results: NRP1 expression was downregulated in podocytes by the addition of AGE-BSA. In contrast, phorbolester induced NRP1 mRNA and protein expression. The wild-type promoter transcriptional activity was significantly reduced when podocytes were treated with AGE-BSA compared with control, unmodified BSA. Point mutations of proximal and distal Sp1-like sites inhibited basal NRP1 promoter activity. AGE-BSA failed to further suppress transcriptional activity of these constructs. Double mutation of the Sp1A and Sp1B binding sites completely abolished NRP1 transcriptional activity. Gel shift analysis showed a specific binding of the Sp1 transcription factor. Treatment of podocytes with AGE-BSA revealed a decrease in Sp1 binding to consensus sequences, but no effect on AP1 binding. Conclusions: AGE-BSA inhibits NRP1 promoter transcriptional activity in podocytes by reducing the binding ability of the Sp1 transcription factor to attach to the NRP1 promoter.

Keywords:
Advanced glycation end products, Podocytes, Neuropilin-1 promoter, Diabetic nephropathy
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2009
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