Antinociceptive Effects of Lipid Raft Disruptors, a Novel Carboxamido-Steroid and Methyl β-Cyclodextrin, in Mice by Inhibiting Transient Receptor Potential Vanilloid 1 and Ankyrin 1 Channel Activation

Ádám Horváth,Zsuzsanna Helyes,Tünde Biró-Sütő,Maja Payrits,Rita Skoda-Földes,Eszter Szánti-Pintér,Boglárka Kántás,Éva Szőke

doi:10.3389/fphys.2020.559109

Ádám Horváth, Zsuzsanna Helyes + Show 6 more

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https://doi.org/10.3389/fphys.2020.559109

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Abstract

Transient Receptor Potential Vanilloid 1 and Ankyrin 1 (TRPV1, TRPA1) cation channels are expressed in nociceptive primary sensory neurons, and play an integrative role in pain processing and inflammatory functions. Lipid rafts are liquid-ordered plasma membrane microdomains rich in cholesterol, sphingomyelin, and gangliosides. We earlier proved that lipid raft disintegration by cholesterol depletion using a novel carboxamido-steroid compound (C1) and methyl β-cyclodextrin (MCD) significantly and concentration-dependently inhibit TRPV1 and TRPA1 activation in primary sensory neurons and receptor-expressing cell lines. Here we investigated the effects of C1 compared to MCD in mouse pain models of different mechanisms. Both C1 and MCD significantly decreased the number of the TRPV1 activation (capsaicin)-induced nocifensive eye-wiping movements in the first hour by 45% and 32%, respectively, and C1 also in the second hour by 26%. Furthermore, C1 significantly decreased the TRPV1 stimulation (resiniferatoxin)-evoked mechanical hyperalgesia involving central sensitization processes, while its inhibitory effect on thermal allodynia was not statistically significant. In contrast, MCD did not affect these resiniferatoxin-evoked nocifensive responses. Both C1 and MCD had inhibitory action on TRPA1 activation (formalin)-induced acute nocifensive reactions (paw liftings, lickings, holdings, and shakings) in the second, neurogenic inflammatory phase by 36% and 51%, respectively. These are the first in vivo data showing that our novel lipid raft disruptor carboxamido-steroid compound exerts antinociceptive and antihyperalgesic effects by inhibiting TRPV1 and TRPA1 ion channel activation similarly to MCD, but in 150-fold lower concentrations. It is concluded that C1 is a useful experimental tool to investigate the effects of cholesterol depletion in animal models, and it also might open novel analgesic drug developmental perspectives.

Highlights

We present here the analgesic effect of lipid raft decomposition depleting cholesterol by carboxamido-steroid compound (C1) and Methyl β-cyclodextrin (MCD) (Szánti-Pintér et al, 2015; Sághy et al, 2018)
We demonstrated that C1 and MCD diminished Transient Receptor Potential Vanilloid 1 (TRPV1) and Transient Receptor Potential Ankyrin 1 (TRPA1) activationinduced acute nocifensive behaviors, C1 inhibited the development of TRPV1 stimulation-evoked mechanical hyperalgesia
Both C1 and MCD significantly diminished the number of CAPS instillation-induced eye-wiping movements in the 1st h by 45 and 32%, respectively, and C1 in the 2nd h by 26%

Summary

Introduction

Transient Receptor Potential (TRP) Vanilloid 1 and Ankyrin 1 (TRPV1 and TRPA1) cation channels are multisteric receptors activated by a variety of inflammatory mediators and tissue irritants, temperature changes and mechanical stimuli besides the classical exogenous agonists such as capsaicin (CAPS), resiniferatoxin (RTX) and formaldehyde, allyl-isothiocyanate (in mustard oil), respectively (McKemy et al, 2002; Peier et al, 2002; Reid and Flonta, 2002; Grimm et al, 2003, 2005; Lee et al, 2003; Bandell et al, 2004; Corey et al, 2004; Jordt et al, 2004; Macpherson et al, 2005, 2007; McNamara et al, 2007; Trevisani et al, 2007; Wagner et al, 2008; Majeed et al, 2010; Vilceanu and Stucky, 2010; Vriens et al, 2011, 2014; Bautista et al, 2013; Drews et al, 2014; Oberwinkler and Philipp, 2014). Pro-inflammatory neuropeptides such as Substance P and calcitonin gene-related peptide released from the activated CAPS-sensitive sensory nerve fibers evoke vasodilation, plasma protein extravasation and inflammatory cells activation in the innervated area called neurogenic inflammation, as well as nociceptor sensitization (Helyes et al, 2003a, 2009; Szolcsányi, 2004) Both TRPV1 and TRPA1 have been in the focus of analgesic and anti-inflammatory drug development, especially for the treatment of chronic neuropathic pain and inflammatory diseases with neurogenic inflammatory components (chronic obstructive pulmonary diseases, psoriasis, arthritis, inflammatory bowel diseases) (Moran et al, 2011; Kaneko and Szallasi, 2014; Nilius and Szallasi, 2014). TRPA1 is considered to Abbreviations: C1, carboxamido-steroid compound; CAPS, capsaicin; CRAC, Cholesterol Recognition/interaction Amino acid Consensus; DHEA, dehydroepiandrosterone; DRG, dorsal root ganglion; E2, 17-β estradiol; MCD, methyl β-cyclodextrin; PGE2, prostaglandin E2; PI(4,5)P2, phosphatidylinositol 4,5-bisphosphate; PS, pregnenolone sulfate; RAMEB, random methylated β-cyclodextrins; RTX, resiniferatoxin; TrkA, tropomyosin-related kinase A; TRP, Transient Receptor Potential; TRPA1, Transient Receptor Potential Ankyrin 1; TRPC5, Transient Receptor Potential Canonical 5; TRPM3, Transient Receptor Potential Melastatin 3; TRPM8, Transient Receptor Potential Melastatin 8; TRPV1, Transient Receptor Potential Vanilloid 1

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