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Chloride channel blockers decrease intracellular pH in cultured renal epithelial LLC-PK1 cells

C D Brown1, A J Dudley

  • 1Department of Physiological Sciences, Medical School, University of Newcastle upon Tyne.

British Journal of Pharmacology
|June 1, 1996
PubMed
Summary
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Chloride channel blockers significantly alter intracellular pH in kidney cells. This study investigated how common blockers like NPPB and niflumic acid affect pH, revealing dose-dependent effects crucial for understanding cellular function.

Area of Science:

  • Cell Biology
  • Renal Physiology
  • Ion Channel Research

Background:

  • Intracellular pH (pHi) regulation is vital for renal epithelial cell function.
  • Chloride channels play a role in maintaining cellular pH balance.
  • Understanding the impact of channel blockers on pHi is essential for interpreting experimental results.

Purpose of the Study:

  • To investigate the effects of various chloride channel blockers on intracellular pH (pHi) in LLC-PK1 renal epithelial cells.
  • To determine the dose-dependent relationship between blocker concentration and pHi changes.
  • To assess the impact of commonly used blockers on pHi at typical experimental concentrations.

Main Methods:

  • LLC-PK1 renal epithelial cell monolayers were utilized.
  • Intracellular pH (pHi) was measured following the addition of chloride channel blockers.

Related Experiment Videos

  • Specific blockers tested included 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB), niflumic acid, flufenamate, diphenylamine-2-carboxylate (DPC), and 4,4'-diisothiocyanatostilbene-2-2'disulphonic acid (DIDS).
  • Dose-response curves were generated to determine apparent K0.5 values for effective blockers.

    • Main Results:

    • Significant intracellular acidification was observed with NPPB, niflumic acid, flufenamate, and DPC.
    • DIDS did not induce significant changes in pHi.
    • The effects were dose-dependent, with apparent K0.5 values determined for niflumic acid (16.7 microM), flufenamate (34.2 microM), and DPC (740 microM).

    Conclusions:

    • Chloride channel blockers, at concentrations used for channel inhibition, exert substantial effects on intracellular pH.
    • Niflumic acid, flufenamate, and DPC cause significant intracellular acidification in renal epithelial cells.
    • These findings highlight the importance of considering the impact of chloride channel blockers on pHi when studying renal cell physiology.