Bupivacaine-induced cellular entry of QX-314 and its contribution to differential nerve block

Br J Pharmacol. 2014 Jan;171(2):438-51. doi: 10.1111/bph.12466.

Abstract

Background and purpose: Selective nociceptor fibre block is achieved by introducing the cell membrane impermeant sodium channel blocker lidocaine N-ethyl bromide (QX-314) through transient receptor potential V1 (TRPV1) channels into nociceptors. We screened local anaesthetics for their capacity to activate TRP channels, and characterized the nerve block obtained by combination with QX-314.

Experimental approach: We investigated TRP channel activation in dorsal root ganglion (DRG) neurons by calcium imaging and patch-clamp recordings, and cellular QX-314 uptake by MS. To characterize nerve block, compound action potential (CAP) recordings from isolated nerves and behavioural responses were analysed.

Key results: Of the 12 compounds tested, bupivacaine was the most potent activator of ruthenium red-sensitive calcium entry in DRG neurons and activated heterologously expressed TRPA1 channels. QX-314 permeated through TRPA1 channels and accumulated intracellularly after activation of these channels. Upon sciatic injections, QX-314 markedly prolonged bupivacaine's nociceptive block and also extended (to a lesser degree) its motor block. Bupivacaine's blockade of C-, but not A-fibre, CAPs in sciatic nerves was extended by co-application of QX-314. Surprisingly, however, this action was the same in wild-type, TRPA1-knockout and TRPV1/TRPA1-double knockout mice, suggesting a TRP-channel independent entry pathway. Consistent with this, high doses of bupivacaine promoted a non-selective, cellular uptake of QX-314.

Conclusions and implications: Bupivacaine, combined with QX-314, produced a long-lasting sensory nerve block. This did not require QX-314 permeation through TRPA1, although bupivacaine activated these channels. Regardless of entry pathway, the greatly extended duration of block produced by QX-314 and bupivacaine may be clinically useful.

Keywords: CAP; DRG; QX-314; TRPA1; TRPV1; nociception; sciatic nerve.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anesthetics, Local / administration & dosage
  • Anesthetics, Local / pharmacology*
  • Animals
  • Behavior, Animal / drug effects
  • Bupivacaine / administration & dosage
  • Bupivacaine / pharmacology*
  • Calcium / metabolism
  • Cell Line
  • Foot Injuries
  • Ganglia, Spinal / drug effects
  • Ganglia, Spinal / metabolism
  • Injections
  • Lidocaine / analogs & derivatives*
  • Lidocaine / metabolism
  • Male
  • Mice, Knockout
  • Nerve Block*
  • Patch-Clamp Techniques
  • Peripheral Nerves / drug effects
  • Primary Cell Culture
  • Rats
  • Rats, Sprague-Dawley
  • Sciatic Nerve / drug effects
  • Sodium Channel Blockers / metabolism*
  • TRPA1 Cation Channel
  • Transient Receptor Potential Channels / genetics
  • Transient Receptor Potential Channels / metabolism

Substances

  • Anesthetics, Local
  • Sodium Channel Blockers
  • TRPA1 Cation Channel
  • Transient Receptor Potential Channels
  • Trpa1 protein, mouse
  • QX-314
  • Lidocaine
  • Calcium
  • Bupivacaine