Zerumbone Modulates α2A-Adrenergic, TRPV1, and NMDA NR2B Receptors Plasticity in CCI-Induced Neuropathic Pain In Vivo and LPS-Induced SH-SY5Y Neuroblastoma In Vitro Models.

Mark R. Hutchinson,Enoch Kumar Perimal,Noor Aishah Mohammed Izham,Jasmine Siew Min Chia,Ahmad Akira Omar Farouk,Mohd Roslan Sulaiman,Sanam Mustafa

doi:10.3389/fphar.2020.00092

Abstract

Zerumbone has shown great potential in various pathophysiological models of diseases, particularly in neuropathic pain conditions. Further understanding the mechanisms of action is important to develop zerumbone as a potential anti-nociceptive agent. Numerous receptors and pathways function to inhibit and modulate transmission of pain signals. Previously, we demonstrated involvement of the serotonergic system in zerumbone’s anti-neuropathic effects. The present study was conducted to determine zerumbone’s modulatory potential involving noradrenergic, transient receptor potential vanilloid type 1 (TRPV1) and N-methyl-D-aspartate (NMDA) receptors in chronic constriction injury (CCI)-induced in vitro and lipopolysaccharide (LPS)-induced SH-SY5Y in vitro neuroinflammatory models. von Frey filament and Hargreaves plantar tests were used to assess allodynia and hyperalgesia in the chronic constriction injury-induced neuropathic pain mouse model. Involvement of specific adrenoceptors were investigated using antagonists— prazosin (α1-adrenoceptor antagonist), idazoxan (α2-adrenoceptor antagonist), metoprolol (β1-adrenoceptor antagonist), ICI 118,551 (β2-adrenoceptor antagonist), and SR 59230 A (β3-adrenoceptor antagonist), co-administered with zerumbone (10 mg/kg). Involvement of excitatory receptors; TRPV and NMDA were conducted using antagonists capsazepine (TRPV1 antagonist) and memantine (NMDA antagonist). Western blot was conducted to investigate the effect of zerumbone on the expression of α2A-adrenoceptor, TRPV1 and NMDA NR2B receptors in CCI-induced whole brain samples of mice as well as in LPS-induced SH-SY5Y neuroblastoma cells. Pre-treatment with α1- and α2-adrenoceptor antagonists significantly attenuated both anti-allodynic and anti-hyperalgesic effects of zerumbone. For β-adrenoceptors, only β2-adrenoceptor antagonist significantly reversed the anti-allodynic and anti-hyperalgesic effects of zerumbone. β1-adrenoceptor antagonist only reversed the anti-allodynic effect of zerumbone. The anti-allodynic and anti-hyperalgesic effects of zerumbone were both absent when TRPV1 and NMDA receptors were antagonized in both nociceptive assays. Zerumbone treatment markedly decreased the expression of α2A-adrenoceptor, while an up-regulation was observed of NMDA NR2B receptors. Expression of TRPV1 receptors however did not significantly change. The in vitro study, representing a peripheral model, demonstrated the reduction of both NMDA NR2B and TRPV1 receptors while significantly increasing α2A-adrenoceptor expression in contrast to the brain samples. Our current findings suggest that the α1-, α2-, β1- and β2-adrenoceptors, TRPV1 and NMDA NR2B are essential for the anti-allodynic and antihyperalgesic effects of zerumbone. Alternatively, we demonstrated the plasticity of these receptors through their response to zerumbone’s administration.

Highlights

IntroductionWe have reported the anti-allodynic and antihyperalgesic properties of zerumbone in a neuropathic pain mouse model (Zulazmi et al, 2015; Chia et al, 2016; Zulazmi et al, 2017)
Zingiber zerumbet (Z. zerumbet) Smith is a wild ginger plant species that has long been used as traditional medicine in Southeast Asia
Administration of antagonists alone did not significantly affect allodynia and hyperalgesia induced by CCI (Figure 1, Figure 2)

Summary

Introduction

We have reported the anti-allodynic and antihyperalgesic properties of zerumbone in a neuropathic pain mouse model (Zulazmi et al, 2015; Chia et al, 2016; Zulazmi et al, 2017). The prevalence of neuropathic pain in the society is increasing at a worrying rate. A pain condition due to lesions or diseases that affect the somatosensory nervous system give rise to neuropathic pain (Merskey, 1986). This debilitating chronic pain condition is common to those who suffer from diabetes, tumor nerve compression, viruses (HIV, varicella zoster virus), central nervous system disorders (multiple sclerosis, stroke), and surgical procedures (Baron and Tolle, 2008; Jensen et al, 2009)

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