Anti-hyperalgesic effects of a novel TRPM8 agonist in neuropathic rats: a comparison with topical menthol

Pain. 2014 Oct;155(10):2097-107. doi: 10.1016/j.pain.2014.07.022. Epub 2014 Jul 30.


Menthol has historically been used topically to alleviate various pain conditions. At low concentrations, this non-selective TRPM8 agonist elicits a cooling sensation, however higher concentrations result in cold hyperalgesia in normal subjects and paradoxically analgesia in neuropathic patients. Through behavioural and electrophysiological means, we examined whether this back-translated into a pre-clinical rodent model. Menthol was applied topically to the hind paws of naive and spinal nerve-ligated (SNL) rats. In behavioural assays, menthol did not affect withdrawal thresholds to mechanical stimulation and 10% and 40% menthol rarely sensitised withdrawals to innocuous cooling in naïve rats. However, in SNL rats, 10% and 40% menthol alleviated cold hypersensitivity. This was partly corroborated by in vivo electrophysiological recordings of dorsal horn lamina V/VI neurones. As several studies have implicated TRPM8 in analgesia, we examined whether a novel systemically available TRPM8 agonist, M8-Ag, had more potent anti-hyperalgesic effects than menthol in neuropathic rats. In vitro, M8-Ag activates TRPM8, expressed in HEK293 cells, with an EC50 of 44.97 nM. In vivo, M8-Ag inhibited neuronal responses to innocuous and noxious cooling in SNL rats with no effect in sham-operated rats. This effect was modality selective; M8-Ag did not alter neuronal responses to mechanical, heat or brush stimulation. In addition, M8-Ag attenuated behavioural hypersensitivity to innocuous cooling but not mechanical stimulation. These data suggest that menthol induced hyperalgesia is not consistently replicable in the rat and that the analgesic properties are revealed by injury. Systemic TRPM8 agonists might be beneficial in neuropathy without affecting normal cold sensitivity.

Keywords: Cold hyperalgesia; Dorsal horn; In vivo electrophysiology; Menthol; Neuropathy; TRPM8.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Analgesics / administration & dosage
  • Analgesics / therapeutic use*
  • Animals
  • Cold Temperature
  • Disease Models, Animal
  • Hyperalgesia / drug therapy*
  • Hyperalgesia / etiology
  • Male
  • Menthol / administration & dosage
  • Menthol / therapeutic use*
  • Morpholines / agonists*
  • Neuralgia / drug therapy*
  • Neuralgia / etiology
  • Pain Threshold / drug effects
  • Peripheral Nerve Injuries / complications
  • Rats
  • Rats, Sprague-Dawley
  • TRPM Cation Channels / agonists*
  • Triazoles / agonists*


  • (4-(5-(4-chlorophenyl)-4-phenyl-4H-1,2,4-triazol-3-yl)morpholine
  • Analgesics
  • Morpholines
  • TRPM Cation Channels
  • Triazoles
  • Trpm8 protein, rat
  • Menthol