Buffer-dependent regulation of aquaporin-1 expression and function in human peritoneal mesothelial cells
BackgroundBiocompatible 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.MethodsHuman peritoneal mesothelial cells (HP...
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| Hauptverfasser: | , , , , , , , , |
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| Dokumenttyp: | Article (Journal) |
| Sprache: | Englisch |
| Veröffentlicht: |
1 March 2012
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| In: |
Pediatric nephrology
Year: 2012, Jahrgang: 27, Heft: 7, Pages: 1165-1177 |
| ISSN: | 1432-198X |
| DOI: | 10.1007/s00467-012-2120-1 |
| Online-Zugang: | Verlag, Volltext: http://dx.doi.org/10.1007/s00467-012-2120-1 Verlag, Volltext: https://link.springer.com/article/10.1007/s00467-012-2120-1 
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| Verfasserangaben: | Yihui Zhai, Jacek Bloch, Meike Hömme, Julia Schaefer, Thilo Hackert, Bärbel Philippin, Vedat Schwenger, Franz Schaefer, Claus P. Schmitt |
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| 520 | |a BackgroundBiocompatible 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.MethodsHuman 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.ResultsIncubation 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.ConclusionsPeritoneal 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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