NOX4 is an early initiator of neuropathic pain

Exp Neurol. 2017 Feb:288:94-103. doi: 10.1016/j.expneurol.2016.11.008. Epub 2016 Nov 14.

Abstract

Treatment of neuropathic pain remains challenging as the etiology is heterogeneous and pathomechanisms are incompletely understood. One possible mechanism is oxidative stress due to unphysiological reactive oxygen species (ROS) formation. The only know dedicated enzymatic source of ROS are NADPH oxidases of which the type 4 isoform (NOX4) has been suggested to be involved in the subacute and chronic phase of neuropathic pain. Here, we aim to translate this finding into a treatment strategy by examining the efficacy of the NOX1/4-specific inhibitor GKT136901 using the chronic constriction injury (CCI) mouse model of neuropathic pain. Unexpectedly, post-nerve lesion treatment using GKT136901 was ineffective to reduce pain-related behavior after CCI. We therefore re-investigated the role of NOX4 using an independent KO mouse model. Early after CCI we found an increase in pro-inflammatory cytokines, ROS formation and the oxidative stress marker nitrotyrosine in the lesioned nerve together with an upregulated Nox4 gene expression. In NOX4 KO mice, mechanical allodynia was markedly reduced from day 4 after nerve injury as were all ROS related and acute biomarkers. In addition, we observed a reduction in the CCI-induced upregulation of pro-inflammatory cytokines in the sciatic nerve and dorsal root ganglia along with NOX4-deficiency. Thus, we conclude that NOX4 is involved in the development of neuropathic pain states by producing oxidative stress and subsequent cytokine dysregulation at the lesion site. This appears at very early stages immediately after nerve injury explaining ineffectiveness of post-acute pharmacological NOX inhibition. We suggest that future target validation of NOX4 should now focus on defining the possible therapeutic window in human neuropathic pain.

Keywords: Chronic constriction injury; GKT136901; NOX4; Neuropathic pain.

MeSH terms

  • Analysis of Variance
  • Animals
  • Cytokines / metabolism
  • Disease Models, Animal
  • Ganglia, Spinal / metabolism
  • Ganglia, Spinal / pathology
  • Gene Expression Regulation / drug effects*
  • Gene Expression Regulation / genetics
  • Hyperalgesia / drug therapy
  • Hyperalgesia / physiopathology*
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • NADPH Oxidase 4
  • NADPH Oxidases / genetics
  • NADPH Oxidases / metabolism*
  • Neuralgia / drug therapy
  • Neuralgia / metabolism*
  • Neuralgia / pathology
  • Neuralgia / physiopathology*
  • Pain Measurement
  • Pyrazoles / therapeutic use
  • Pyridones / therapeutic use
  • RNA, Messenger / metabolism
  • Reactive Oxygen Species / metabolism
  • Sciatic Nerve / pathology

Substances

  • 2-(2-chlorophenyl)-4-methyl-5-(pyridin-2-ylmethyl)-1H-pyrazolo(4,3-c)pyridine-3,6(2H,5H)-dione
  • Cytokines
  • Pyrazoles
  • Pyridones
  • RNA, Messenger
  • Reactive Oxygen Species
  • NADPH Oxidase 4
  • NADPH Oxidases
  • Nox4 protein, mouse