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

Pediatr Nephrol. 2012 Jul;27(7):1165-77. doi: 10.1007/s00467-012-2120-1. Epub 2012 Mar 1.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aquaporin 1 / biosynthesis*
  • Bicarbonates / pharmacology
  • Blotting, Western
  • Buffers*
  • Cell Movement / drug effects
  • Cells, Cultured
  • Epithelial Cells / drug effects*
  • Epithelial Cells / metabolism
  • Epithelium / drug effects
  • Epithelium / metabolism
  • Hemodialysis Solutions / pharmacology*
  • Humans
  • Lactic Acid / pharmacology
  • Peritoneal Dialysis / adverse effects*
  • Peritoneum / cytology
  • Peritoneum / drug effects
  • Peritoneum / metabolism
  • RNA, Messenger
  • RNA, Small Interfering
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • AQP1 protein, human
  • Bicarbonates
  • Buffers
  • Hemodialysis Solutions
  • RNA, Messenger
  • RNA, Small Interfering
  • Aquaporin 1
  • Lactic Acid