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The role of dorsal root ganglia alpha-7 nicotinic acetylcholine receptor in complete Freund’s adjuvant-induced chronic inflammatory pain

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Abstract

Background

Alpha-7 nicotinic acetylcholine receptor (α7 nAChR) was reported to have a critical role in the regulation of pain sensitivity and neuroinflammation. However, the expression level of α7 nAChR in dorsal root ganglion (DRG) and the underlying neuroinflammatory mechanisms associated with hyperalgesia are still unknown.

Methods

In the present study, the expression and mechanism of α7 nAChR in chronic inflammatory pain was investigated using a complete Freund’s adjuvant (CFA)-induced chronic inflammatory pain model. Subsequently, a series of assays including immunohistochemistry, western blotting, and quantitative real-time polymerase chain reaction (qRT-PCR) were performed.

Results

α7 nAChR was mostly colocalized with NeuN in DRG and upregulated after CFA injection. Microinjection of α7 nAChR siRNA into ipsilateral L4/5 DRGs aggravated the CFA-induced pain hypersensitivity. Intrathecal α7 nAChR agonist GTS-21 attenuated the development of CFA-induced mechanical and temperature-related pain hypersensitivities. In neuronal the SH-SY5Y cell line, the knockdown of α7 nAChRs triggered the upregulation of TRAF6 and NF-κB under CFA-induced inflammatory conditions, while agitation of α7 nAChR suppressed the TRAF6/NF-κB activation. α7 nAChR siRNA also exacerbated the secretion of pro-inflammatory mediators from LPS-induced SH-SY5Y cells. Conversely, α7 nAChR-specific agonist GTS-21 diminished the release of interleukin-1beta (IL-1β), IL-6, IL-8, and tumor necrosis factor-α (TNFα) in SH-SY5Y cells under inflammatory conditions. Mechanistically, the modulation of pain sensitivity and neuroinflammatory action of α7 nAChR may be mediated by the TRAF6/NF-κB signaling pathway.

Conclusions

The findings of this study suggest that α7 nAChR may be potentially utilized as a therapeutic target for therapeutics of chronic inflammatory pain.

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References

  • Abbas M, Alzarea S, Papke RL, Rahman S (2019) The α7 nicotinic acetylcholine receptor positive allosteric modulator prevents lipopolysaccharide-induced allodynia, hyperalgesia and TNF-α in the hippocampus in mice. Pharmacol Rep 71:1168–1176

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Albers KM, Zhang XL, Diges CM, Schwartz ES, Yang CI, Davis BM, Gold MS (2014) Artemin growth factor increases nicotinic cholinergic receptor subunit expression and activity in nociceptive sensory neurons. Mol Pain 10:31

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Alsharari SD, Freitas K, Damaj MI (2013) Functional role of alpha7 nicotinic receptor in chronic neuropathic and inflammatory pain: studies in transgenic mice. Biochem Pharmacol 86:1201–1207

    Article  CAS  PubMed  Google Scholar 

  • Bagdas D, Wilkerson JL, Kulkarni A, Toma W, Alsharari S, Gul Z, Lichtman AH, Papke RL, Thakur GA, Damaj MI (2016) The α7 nicotinic receptor dual allosteric agonist and positive allosteric modulator GAT107 reverses nociception in mouse models of inflammatory and neuropathic pain. Br J Pharmacol 173:2506–2520

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bagdas D, Gurun MS, Flood P, Papke RL, Damaj MI (2018) New insights on neuronal nicotinic acetylcholine receptors as targets for pain and inflammation: a focus on α7 nAChRs. Curr Neuropharmacol 16:415–425

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bang S, Donnelly CR, Luo X, Toro-Moreno M, Tao X, Wang Z, Chandra S, Bortsov AV, Derbyshire ER, Ji RR (2021) Activation of GPR37 in macrophages confers protection against infection-induced sepsis and pain-like behaviour in mice. Nat Commun 12:1704

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Bao J, Liu Y, Yang J, Gao Q, Shi SQ, Garfield RE, Liu H (2016) Nicotine inhibits LPS-induced cytokine production and leukocyte infiltration in rat placenta. Placenta 39:77–83

    Article  CAS  PubMed  Google Scholar 

  • Baron EP, Lucas P, Eades J, Hogue O (2018) Patterns of medicinal cannabis use, strain analysis, and substitution effect among patients with migraine, headache, arthritis, and chronic pain in a medicinal cannabis cohort. J Headache Pain 19:37

    Article  PubMed  PubMed Central  Google Scholar 

  • Busse JW, Wang L, Kamaleldin M, Craigie S, Riva JJ, Montoya L, Mulla SM, Lopes LC, Vogel N, Chen E, Kirmayr K, de Oliveira K, Olivieri L, Kaushal A, Chaparro LE, Oyberman I, Agarwal A, Couban R, Tsoi L, Lam T, Vandvik PO, Hsu S, Bala MM, Schandelmaier S, Scheidecker A, Ebrahim S, Ashoorion V, Rehman Y, Hong PJ, Ross S, Johnston BC, Kunz R, Sun X, Buckley N, Sessler DI, Guyatt GH (2018) Opioids for chronic noncancer pain: a systematic review and meta-analysis. JAMA 320:2448–2460

    Article  PubMed  PubMed Central  Google Scholar 

  • Chen Y, Chen X, Yu J, Xu X, Wei X, Gu X, Liu C, Zhang D, Xu Z (2016) JAB1 is involved in neuropathic pain by regulating JNK and NF-κB activation after chronic constriction injury. Neurochem Res 41:1119–1129

    Article  CAS  PubMed  Google Scholar 

  • Chernyavsky AI, Arredondo J, Skok M, Grando SA (2010) Auto/paracrine control of inflammatory cytokines by acetylcholine in macrophage-like U937 cells through nicotinic receptors. Int Immunopharmacol 10:308–315

    Article  CAS  PubMed  Google Scholar 

  • Chrestia JF, Bruzzone A, Esandi MDC, Bouzat C (2021) Tyrosine phosphorylation differentially fine-tunes ionotropic and metabotropic responses of human α7 nicotinic acetylcholine receptor. Cell Mol Life Sci 78:5381–5395

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Criscuolo C, Accorroni A, Domenici L, Origlia N (2015) Impaired synaptic plasticity in the visual cortex of mice lacking α7-nicotinic receptor subunit. Neuroscience 294:166–171

    Article  CAS  PubMed  Google Scholar 

  • Debar L, Benes L, Bonifay A, Deyo RA, Elder CR, Keefe FJ, Leo MC, McMullen C, Mayhew M, Owen-Smith A, Smith DH, Trinacty CM, Vollmer WM (2018) Interdisciplinary team-based care for patients with chronic pain on long-term opioid treatment in primary care (PPACT)—protocol for a pragmatic cluster randomized trial. Contemp Clin Trials 67:91–99

    Article  PubMed  PubMed Central  Google Scholar 

  • Donvito G, Bagdas D, Toma W, Rahimpour E, Jackson A, Meade JA, Alsharari S, Kulkarni AR, Ivy Carroll F, Lichtman AH, Papke RL, Thakur GA, Imad Damaj M (2017) The interaction between alpha 7 nicotinic acetylcholine receptor and nuclear peroxisome proliferator-activated receptor-α represents a new antinociceptive signaling pathway in mice. Exp Neurol 295:194–201

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ge YT, Zhong AQ, Xu GF, Lu Y (2019) Resveratrol protects BV2 mouse microglial cells against LPS-induced inflammatory injury by altering the miR-146a-5p/TRAF6/NF-κB axis. Immunopharmacol Immunotoxicol 41:549–557

    Article  CAS  PubMed  Google Scholar 

  • Ghasemlou N, Chiu IM, Julien JP, Woolf CJ (2015) CD11b+Ly6G- myeloid cells mediate mechanical inflammatory pain hypersensitivity. Proc Natl Acad Sci USA 112:E6808–E6817

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Godin JR, Roy P, Quadri M, Bagdas D, Toma W, Narendrula-Kotha R, Kishta OA, Damaj MI, Horenstein NA, Papke RL, Simard AR (2019) A silent agonist of α7 nicotinic acetylcholine receptors modulates inflammation ex vivo and attenuates EAE. Brain Behav Immun 87:286–300

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Huang LN, Zou Y, Wu SG, Zhang HH, Mao QX, Li JB, Tao YX (2019) Fn14 participates in neuropathic pain through NF-κB pathway in primary sensory neurons. Mol Neurobiol 56:7085–7096

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kalkman HO, Feuerbach D (2016) Modulatory effects of α7 nAChRs on the immune system and its relevance for CNS disorders. Cell Mol Life Sci 73:2511–2530

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Khan MA, Farkhondeh M, Crombie J, Jacobson L, Kaneki M, Martyn JA (2012) Lipopolysaccharide upregulates α7 acetylcholine receptors: stimulation with GTS-21 mitigates growth arrest of macrophages and improves survival in burned mice. Shock 38:213–219

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Khan S, Choi RJ, Lee J, Kim YS (2016) Attenuation of neuropathic pain and neuroinflammatory responses by a pyranocoumarin derivative, anomalin in animal and cellular models. Eur J Pharmacol 774:95–104

    Article  CAS  PubMed  Google Scholar 

  • Kim B, Lee KY, Park B (2018) Icariin abrogates osteoclast formation through the regulation of the RANKL-mediated TRAF6/NF-κB/ERK signaling pathway in Raw264.7 cells. Phytomedicine 51:181–190

    Article  CAS  PubMed  Google Scholar 

  • Liang L, Fan L, Tao B, Yaster M, Tao YX (2013) Protein kinase B/Akt is required for complete Freund’s adjuvant-induced upregulation of Nav1.7 and Nav1.8 in primary sensory neurons. J Pain 14:638–647

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Licheri V, Lagstr MO, Lotfi A, Patton MH, Wigstr MH, Mathur B, Adermark L (2018) Complex control of striatal neurotransmission by nicotinic acetylcholine receptors via excitatory inputs onto medium spiny neurons. J Neurosci 38:6597–6607

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Liu GJ, Zhang QR, Gao X, Wang H, Tao T, Gao YY, Zhou Y, Chen XX, Li W, Hang CH (2020) MiR-146a ameliorates hemoglobin-induced microglial inflammatory response via TLR4/IRAK1/TRAF6 associated pathways. Front Neurosci 14:311

    Article  PubMed  PubMed Central  Google Scholar 

  • Loram LC, Harrison JA, Chao L, Taylor FR, Reddy A, Travis CL, Giffard R, Al-Abed Y, Tracey K, Maier SF, Watkins LR (2010) Intrathecal injection of an alpha seven nicotinic acetylcholine receptor agonist attenuates gp120-induced mechanical allodynia and spinal pro-inflammatory cytokine profiles in rats. Brain Behav Immun 24:959–967

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Maiar M, Leese C, Certo M, Echeverria-Altuna I, Mangione AS, Arsenault J, Davletov B, Hunt SP (2018) Selective neuronal silencing using synthetic botulinum molecules alleviates chronic pain in mice. Sci Transl Med 10:eaar7384

  • Matsuoka Y, Yamashita A, Matsuda M, Kawai K, Sawa T, Amaya F (2019) NLRP2 inflammasome in dorsal root ganglion as a novel molecular platform that produces inflammatory pain hypersensitivity. Pain 160:2149–2160

    Article  CAS  PubMed  Google Scholar 

  • Mengke NS, Hu B, Han QP, Deng YY, Fang M, Xie D, Li A, Zeng HK (2016) Rapamycin inhibits lipopolysaccharide-induced neuroinflammation in vitro and in vivo. Mol Med Rep 14:4957–4966

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Moschetti G, Amodeo G, Paladini MS, Molteni R, Balboni G, Panerai A, Sacerdote P, Franchi S (2019) Prokineticin 2 promotes and sustains neuroinflammation in vincristine treated mice: focus on pain and emotional like behavior. Brain Behav Immun 82:422–431

    Article  CAS  PubMed  Google Scholar 

  • Müller-Schwefe G, Morlion B, Ahlbeck K, Alon E, Coaccioli S, Coluzzi F, Huygen F, Jaksch W, Kalso E, Kocot-Kępska M, Kress HG, Mangas AC, Margarit Ferri C, Mavrocordatos P, Nicolaou A, Hernández CP, Pergolizzi J, Schäfer M, Sichère P (2017) Treatment for chronic low back pain: the focus should change to multimodal management that reflects the underlying pain mechanisms. Curr Med Res Opin 33:1199–1210

    Article  PubMed  Google Scholar 

  • Niranjan R, Nath C, Shukla R (2012) Melatonin attenuated mediators of neuroinflammation and alpha-7 nicotinic acetylcholine receptor mRNA expression in lipopolysaccharide (LPS) stimulated rat astrocytoma cells, C6. Free Radic Res 46:1167–1177

    Article  CAS  PubMed  Google Scholar 

  • Ortmann KL, Chattopadhyay M (2014) Decrease in neuroimmune activation by HSV-mediated gene transfer of TNFα soluble receptor alleviates pain in rats with diabetic neuropathy. Brain Behav Immun 41:144–151

    Article  CAS  PubMed  Google Scholar 

  • Pasquale MK, Dufour R, Schaaf D, Reiners AT, Mardekian J, Joshi AV, Patel NC (2014) Pain conditions ranked by healthcare costs for members of a national health plan. Pain Pract 14:117–131

    Article  PubMed  Google Scholar 

  • Peng H, Zou L, Xie J, Wu H, Wu B, Zhu G, Lv Q, Zhang X, Liu S, Li G, Xu H, Gao Y, Xu C, Zhang C, Wang S, Xue Y, Liang S (2017) lncRNA NONRATT021972 siRNA decreases diabetic neuropathic pain mediated by the P2X(3) receptor in dorsal root ganglia. Mol Neurobiol 54:511–523

    Article  CAS  PubMed  Google Scholar 

  • Pinho-Ribeiro FA, Fattori V, Zarpelon AC, Borghi SM, Staurengo-Ferrari L, Carvalho TT, Alves-Filho JC, Cunha FQ, Cunha TM, Casagrande R, Verri WA (2016) Pyrrolidine dithiocarbamate inhibits superoxide anion-induced pain and inflammation in the paw skin and spinal cord by targeting NF-κB and oxidative stress. Inflammopharmacology 24:97–107

    Article  CAS  PubMed  Google Scholar 

  • Ruan X, Kaye AD (2016) A call for saving interdisciplinary pain management. J Orthop Sports Phys Ther 46:1021–1023

    Article  PubMed  Google Scholar 

  • Sabec MH, Wonnacott S, Warburton EC, Bashir ZI (2018) Nicotinic acetylcholine receptors control encoding and retrieval of associative recognition memory through plasticity in the medial prefrontal cortex. Cell Rep 22:3409–3415

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Shenkarev ZO, Shulepko MA, Bychkov ML, Kulbatskii DS, Shlepova OV, Vasilyeva NA, Andreev-Andrievskiy AA, Popova AS, Lagereva EA, Loktyushov EV, Koshelev SG, Thomsen MS, Dolgikh DA, Kozlov SA, Balaban PM, Kirpichnikov MP, Lyukmanova EN (2020) Water-soluble variant of human Lynx1 positively modulates synaptic plasticity and ameliorates cognitive impairment associated with α7-nAChR dysfunction. J Neurochem 155:45–61

    Article  CAS  PubMed  Google Scholar 

  • Stötzner P, Spahn V, Celik M, Labuz D, Machelska H (2018) Mu-opioid receptor agonist induces Kir3 currents in mouse peripheral sensory neurons—effects of nerve injury. Front Pharmacol 9:1478

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Sun R, Liu Y, Hou B, Lei Y, Bo J, Zhang W, Sun Y, Zhang Y, Zhang Z, Liu Z, Huo W, Mao Y, Ma Z, Gu X (2019) Perioperative activation of spinal α7 nAChR promotes recovery from preoperative stress-induced prolongation of postsurgical pain. Brain Behav Immun 79:294–308

    Article  CAS  PubMed  Google Scholar 

  • van Hecke O, Austin SK, Khan RA, Smith BH, Torrance N (2014) Neuropathic pain in the general population: a systematic review of epidemiological studies. Pain 155:654–662

    Article  PubMed  Google Scholar 

  • Wedn AM, El-Gowilly SM, El-Mas MM (2019) Nicotine reverses the enhanced renal vasodilator capacity in endotoxic rats: role of α7/α4β2 nAChRs and HSP70. Pharmacol Rep 71:782–793

    Article  CAS  PubMed  Google Scholar 

  • Xiang X, Wang S, Shao F, Fang J, Xu Y, Wang W, Sun H, Liu X, Du J (2019) Electroacupuncture stimulation alleviates CFA-induced inflammatory pain via suppressing P2X3 expression. Int J Mol Sci 20:3248

    Article  CAS  PubMed Central  Google Scholar 

  • Yang Y, Tan X, Xu J, Wang T, Liang T, Xu X, Ma C, Xu Z, Wang W, Li H, Shen H, Li X, Dong W, Chen G (2020) Luteolin alleviates neuroinflammation via downregulating the TLR4/TRAF6/NF-κB pathway after intracerebral hemorrhage. Biomed Pharmacother 126:110044

    Article  CAS  PubMed  Google Scholar 

  • Zhai X, Yan Z, Zhao J, Chen K, Yang Y, Cai M, He C, Huang C, Li B, Yang M, Zhou X, Zhao Y, Wei X, Bai Y, Li M (2020) Muscone ameliorates ovariectomy-induced bone loss and receptor activator of nuclear factor-κb ligand-induced osteoclastogenesis by suppressing TNF receptor-associated factor 6-mediated signaling pathways. Front Pharmacol 11:348

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhao JY, Liang L, Gu X, Li Z, Wu S, Sun L, Atianjoh FE, Feng J, Mo K, Jia S, Lutz BM, Bekker A, Nestler EJ, Tao YX (2017) DNA methyltransferase DNMT3a contributes to neuropathic pain by repressing Kcna2 in primary afferent neurons. Nat Commun 8:14712

    Article  PubMed  PubMed Central  Google Scholar 

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Funding

This study was supported by the National Natural Science Foundation of China (No. 81401576); Shanghai Municipal Commission of Health and Family Planning, China (No. 20164Y0263). The authors would like to express their gratitude to EditSprings (https://www.editsprings.com/) for the expert linguistic services provided.

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Authors and Affiliations

  1. Department of Anesthesiology, Shanghai General Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, 200080, China

    Xiaoyu Zhang, Fangxia Xu, Lijuan Wang, Jinbao Li, Jianhai Zhang & Lina Huang

  2. Department of Anesthesiology, International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai, 200030, China

    Xiaoyu Zhang

  3. Shanghai Key Laboratory of Embryo Original Diseases, Shanghai, 200030, China

    Xiaoyu Zhang

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  1. Xiaoyu Zhang
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  2. Fangxia Xu
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  3. Lijuan Wang
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  4. Jinbao Li
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  5. Jianhai Zhang
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  6. Lina Huang
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Contributions

(I) Conception and design: XZ, LH; (II) Administrative support: JL, LH; (III) Provision of study materials or patients: all authors; (IV) Collection and assembly of data: all authors; (V) Data analysis and interpretation: all authors; (VI) Manuscript writing: all authors; (VII) Final approval of manuscript: all authors.

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Correspondence to Jianhai Zhang or Lina Huang.

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The authors have no conflicts of interest to declare.

Ethical statement

The authors are accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved. The animal experiments were approved by the Laboratory Animal Ethics Committee of International Peace Maternity & Child Health Hospital (Shanghai, China).

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10787_2021_873_MOESM1_ESM.tif

Supplementary file1 Supplementary Fig. 1. Effect of intrathecal α7 nAChR agonist GTS-21 on basal responses on the contralateral side. A–C Effect of intrathecal vehicle or 20,000 nM/mice GTS-21 on paw withdrawal frequencies to mechanical stimulation, paw withdrawal latencies to heat, and cold stimuli on 3 day post-CFA injection or sham surgery. n = 6 mice/group. Two-way analysis of variance (ANOVA) followed by Bonferroni post hoc test. D–F Dose-dependent effect of intrathecal GTS-21 or vehicle on paw withdrawal frequency to mechanical stimuli and paw withdrawal latencies to heat and cold stimuli on day 3 post-CFA injection. n = 6 mice/group. Two-way analysis of variance (ANOVA) followed by Bonferroni post hoc test (TIF 1295 kb)

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Zhang, X., Xu, F., Wang, L. et al. The role of dorsal root ganglia alpha-7 nicotinic acetylcholine receptor in complete Freund’s adjuvant-induced chronic inflammatory pain. Inflammopharmacol 29, 1487–1501 (2021). https://doi.org/10.1007/s10787-021-00873-0

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  • DOI: https://doi.org/10.1007/s10787-021-00873-0

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