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Published in: Pediatric Nephrology 7/2012

01-07-2012 | Original Article

Buffer-dependent regulation of aquaporin-1 expression and function in human peritoneal mesothelial cells

Authors: Yihui Zhai, Jacek Bloch, Meike Hömme, Julia Schaefer, Thilo Hackert, Bärbel Philippin, Vedat Schwenger, Franz Schaefer, Claus P. Schmitt

Published in: Pediatric Nephrology | Issue 7/2012

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Abstract

Background

Biocompatible peritoneal dialysis fluids (PDF) are buffered with lactate and/or bicarbonate. We hypothesized that the reduced toxicity of the biocompatible solutions might unmask specific effects of the buffer type on mesothelial cell functions.

Methods

Human peritoneal mesothelial cells (HPMC) were incubated with bicarbonate (B-)PDF or lactate-buffered (L-)PDF followed by messenger RNA (mRNA) and protein analysis. Gene silencing was achieved using small interfering RNA (siRNA), functional studies using Transwell culture systems, and monolayer wound-healing assays.

Results

Incubation with B-PDF increased HPMC migration in the Transwell and monolayer wound-healing assay to 245 ± 99 and 137 ± 11% compared with L-PDF. Gene silencing showed this effect to be entirely dependent on the expression of aquaporin-1 (AQP-1) and independent of AQP-3. Exposure of HPMC to B-PDF increased AQP-1 mRNA and protein abundance to 209 ± 80 and 197 ± 60% of medium control; the effect was pH dependent. L-PDF reduced AQP-1 mRNA. Addition of bicarbonate to L-PDF increased AQP-1 abundance by threefold; mRNA half-life remained unchanged. Immunocytochemistry confirmed opposite changes of AQP-1 cell-membrane abundance with B-PDF and L-PDF.

Conclusions

Peritoneal mesothelial AQP-1 abundance and migration capacity is regulated by pH and buffer agents used in PD solutions. In vivo studies are required to delineate the impact with respect to long-term peritoneal membrane integrity and function.
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Metadata
Title
Buffer-dependent regulation of aquaporin-1 expression and function in human peritoneal mesothelial cells
Authors
Yihui Zhai
Jacek Bloch
Meike Hömme
Julia Schaefer
Thilo Hackert
Bärbel Philippin
Vedat Schwenger
Franz Schaefer
Claus P. Schmitt
Publication date
01-07-2012
Publisher
Springer-Verlag
Published in
Pediatric Nephrology / Issue 7/2012
Print ISSN: 0931-041X
Electronic ISSN: 1432-198X
DOI
https://doi.org/10.1007/s00467-012-2120-1

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