Tadalafil attenuates hypotonicity-induced Ca2+ influx via TRPV2 and TRPV4 in primary rat bladder urothelial cell cultures

Neurourol Urodyn. 2018 Jun;37(5):1541-1548. doi: 10.1002/nau.23423. Epub 2018 Mar 22.

Abstract

Aims: To investigate the localization of phosphodiesterase 5 (PDE5) and the molecular mechanism underlying the effect of the PDE5 inhibitor tadalafil in signal transduction in the bladder urothelium.

Methods: PDE5 expression in rat bladder tissues and cultured primary rat bladder urothelial cells was evaluated using immunochemistry and western blot assays. Ca2+ influx in cells exposed to isotonic solution, hypotonic solution, a selective transient receptor potential vanilloid 2 (TRPV2) channel agonist (cannabidiol), a selective TRPV4 channel agonist (GSK1016790A), a TRP cation channel melastatin 7 (TRPM7) channel agonist (PIP2), or a purinergic receptor agonist (ATP) in the presence or absence of 10 µM tadalafil was evaluated using calcium imaging techniques. We also evaluated stretch-induced changes in ATP concentration in the mouse bladder in the presence or absence of 100 µM tadalafil.

Results: Immunochemistry and western blot analyses demonstrated that PDE5 is abundantly expressed in the bladder urothelium and in primary rat urothelial cells. Ca2+ influx induced by hypotonic stimulation, GSK1016790A, or cannabidiol was significantly inhibited by tadalafil, whereas ATP-induced Ca2+ influx was unaffected by tadalafil. PIP2 did not induce Ca2+ influx. ATP release in tadalafil-pretreated bladders significantly decreased compared to control bladders.

Conclusions: Tadalafil attenuates Ca2+ influx via TRPV4 and TRPV2, and inhibits ATP release in the bladder urothelium. These findings indicate that tadalafil functions as an inhibitor of urothelial signal transduction.

Keywords: ATP; TRP channel; lower urinary tract symptoms; phosphodiesterase type 5; tadalafil.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Animals
  • Calcium / metabolism*
  • Cells, Cultured
  • Hypotonic Solutions
  • Leucine / analogs & derivatives
  • Leucine / pharmacology
  • Male
  • Muscle Tonus / drug effects*
  • Phosphodiesterase 5 Inhibitors / pharmacology*
  • Rats
  • Rats, Sprague-Dawley
  • Signal Transduction / drug effects
  • Sulfonamides / pharmacology
  • TRPM Cation Channels / drug effects
  • TRPV Cation Channels / agonists
  • TRPV Cation Channels / drug effects*
  • Tadalafil / pharmacology*
  • Urinary Bladder / cytology
  • Urinary Bladder / drug effects*
  • Urothelium / cytology
  • Urothelium / drug effects*

Substances

  • Hypotonic Solutions
  • N-(1-((4-(2-(((2,4-dichlorophenyl)sulfonyl)amino)-3-hydroxypropanoyl)-1-piperazinyl)carbonyl)-3-methylbutyl)-1-benzothiophene-2-carboxamide
  • Phosphodiesterase 5 Inhibitors
  • Sulfonamides
  • TRPM Cation Channels
  • TRPV Cation Channels
  • Trpv2 protein, rat
  • Trpv4 protein, rat
  • Tadalafil
  • Adenosine Triphosphate
  • Trpm7 protein, rat
  • Leucine
  • Calcium