Veterinary Research Forum

Effects of thymoquinone on acute corneal and orofacial pains in rats: central involvement of opioid, cannabinoid, muscarinic cholinergic, and serotonin receptors

Document Type : Original Article

Authors

Esmaeal Tamaddonfard
Amir Erfanparast
Afsaneh Niakani

Department of Basic Sciences, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran

https://doi.org/10.30466/vrf.2025.2050017.4622
Abstract
Thymoquinone (TQ), the bioactive compound found in black seed, possesses beneficial properties. In the present study, the effects of oral administration of TQ were investigated on acute corneal and orofacial pains in rats. To clarify the central mechanism of action, muscarinic cholinergic, cannabinergic 1 (CB1) and 5-hydroxytryptamine receptor antagonists were delivered into the 4th ventricle of the brain after oral administration of TQ. Acute corneal and orofacial pains were induced by dropping of hypertonic saline (50.00 µL; 5.00 M) on the corneal surface and subcutaneous injection of capsaicin (1.50 µg; 20.00 µL) in the vibrissal pad, respectively. The eye wiping number and face rubbing duration were recorded as corneal and orofacial pains behavioral responses, respectively. Locomotor activity was measured using an open-field test. The TQ (5.00 mg kg-1) had no effects, while it reduced pain responses at 10.00, 20.00, and 40.00 mg kg-1. Intracerebro-ventricular injections of naloxone (an antagonist of opioid receptors), (N-(Piperidin-1-yl)-5-(4-iodophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide (an antagonist of CB1 receptors), atropine (an antagonist of muscarinic cholinergic receptors), and ondansetron (an antagonist of 5-hydroxytryptamine 3 receptors) at a similar dose of 10.00 µg kg-1 inhibited corneal and orofacial pains suppression caused by 40.00 mg kg-1 TQ. The tested drugs did not affect locomotor activity. It is concluded that TQ caused analgesia in the acute corneal and orofacial pains. Central opioid, cholinergic muscarinic, CB1, and 5-hydroxytryptamine 3 receptors might be involved in the anti-nociceptive effects of TQ.

Keywords

Acute pain
AM251
Atropine
Ondansetron
Thymoquinone
Subjects
1.     Kuner R, Kuner T. Cellular circuits in the brain and their modulation in acute and chronic pain. Physiol Rev 2021; 101(1): 213-258.
2.     Amaechi O, Huffman MM, Featherstone K. Pharmacologic therapy for acute pain. Am Fam Physician 2021; 104(1): 63-72.
3.     Yousofvand N, Moloodi B. An overview of the effect of medicinal herbs on pain. Phytother Res 2023; 37(3): 1057-1081.
4.     Turnaturi R, Piana S, Spoto S, et al. From plant to chemistry: sources of antinociceptive non-opioid active principles for medicinal chemistry and drug design. Molecules 2024; 29(4): 815. doi:10.3390/molecules 29040815.
5.     Aslani MR, Saadat S, Boskabady MH. Comprehensive and updated review on anti-oxidant effects of Nigella sativa and its constituent, thymoquinone, in various disorder. Iran J Basic Med Sci 2024; 27(8): 923-951.
6.     Akarsu GD, Çetin A. The effect of thymoquinone on oxidative stress parameters and apolipoprotein E in Alzheimer model in rats. Dement Geriatr Cogn Disord 2022; 51(4): 297-309.
7.     Xing H, Zhang S, You M, et al. Thymoquinone alleviates paclitaxel-induced peripheral neuropathy through regulation of the TLR4-MyD88 inflammatory pathway. ACS Chem Neurosci 2023; 14(20): 3804-3817.
8.     Goellner E, Rocha CE. Anatomy of trigeminal neuromodulation targets: from periphery to the brain. Prog Neurol Surg 2020; 35: 18-34.
9.     Murray GM, Sessle BJ. Pain-sensorimotor interactions: new perspectives and a new model. Neurobiol Pain 2024; 15: 100150. doi: 10.1016/j.ynpai.2024.100150.
10. Jabbari S, Bananej M, Zarei M, et al. Effects of intrathecal and intracerebroventricular microinjection of kaempferol on pain: possible mechanisms of action. Res Pharm Sci 2021; 16(2): 203-216.
11. Tamaddonfard E, Tamaddonfard S, Pourbaba S. Effects of intra-fourth ventricle injection of crocin on capsaicin-induced orofacial pain in rats. Avicenna J Phytomed 2015; 5(5): 450-457.
12. Naderi S, Tamaddonfard E, Nafisi S, et al. Effect of thymoquinone on acetic acid-induced visceral nociception in rats: role of central cannabinoid and α2-adrenergic receptors. Vet Res Forum 2024; 15(3): 131-138.
13. Tamaddonfard S, Erfanparast A, Tamaddonfard E, et al. The CB1 cannabinoid receptors involvement in anti-epileptic effect of safranal on penicillin-induced epileptiform activity in rats. Vet Res Forum 2024; 15(1): 35-41.
14. Mojtahedin A, Tamaddonfard E, Zanbouri A. Role of central muscarinic cholinergic receptors in the formalin-induced pain in rats. Indian J Pharmacol 2009; 41(3): 144-147.
15. Ramesh ST, Asad M, Dhamanigi SS, et al. Effect of central administration of ondansetron, a 5-hydroxytryptamine-3 receptor antagonist on gastric and duodenal ulcers. Fundam Clin Pharmacol 2009; 23(3): 303-309.
16. Paxinos G, Watson C. The rat brain in stereotaxic coordinates. 6th ed. San Diego, USA: Elsevier Academic Press 2007; 137-140.
17. Meng ID, Barton ST, Goodney I, et al. Progesterone application to the rat forehead produces corneal antinociception. Invest Ophthalmol Vis Sci 2019; 60(5): 1706-1713.
18. Pelissier T, Pajot J, Dallel R. The orofacial capsaicin test in rats: effect of different capsaicin concentrations and morphine. Pain 2002; 96(1-2): 81-87.
19. Farazifard R, Safarpour F, Sheibani V, et al. Eye-wiping test: a sensitive animal model for acute trigeminal pain studies. Brain Res Brain Res Protoc 2005; 16(1-3):44-49.
20. Tamaddonfard E, Erfanparast A, Ghasemi H, et al. The role of histamine H1, H2 and H3 receptors of ventral posteromedial nucleus of thalamus in modulation of trigeminal pain. Eur J Pharmacol 2016: 791: 696-702.
21. Tashiro A, Bereiter DA, Ohta H, et al. Trigeminal sensitization in a closed head model for mild traumatic brain injury. J Neurotrauma 2024; 41(7-8): 985-999.
22. Rita Pereira EM, Souza JM, Carobin NV, et al. Phoneutria toxin PnTx3-5 inhibits TRPV1 channel with antinociceptive action in an orofacial pain model. Neuropharmacology 2020; 162: 107826. doi: 10.1016/ j.neuropharm.2019.107826.
23. Elmaci I, Altinoz MA. Thymoquinone: an edible redox-active quinone for the pharmacotherapy of neuro-degenerative conditions and glial brain tumors. A short review. Biomed Pharmacother 2016; 83: 635-640.
24. Parvardeh S, Sabetkasaei M, Moghimi M, et al. Role of L-arginine/NO/cGMP/KATP channel signaling pathway in the central and peripheral antinociceptive effect of thymoquinone in rats. Iran J Basic Med Sci 2018; 21(6): 625-633.
25. Alenezi SK. The ameliorative effect of thymoquinone on vincristine-induced peripheral neuropathy in mice by modulating cellular oxidative stress and cytokine. Life (Basel) 2022; 13(1): 101. doi:10.3390/life13010101.
26. Sivilotti ML. Flumazenil, naloxone and the 'coma cocktail. Br J Clin Pharmacol 2016: 81(3): 428-436.
27. Corder G, Castro DC, Bruchas MR, et al. Endogenous and exogenous opioids in pain. Annu Rev Neurosci 2018; 41: 453-473.
28. Faouzi A, Varga BR, Majumdar S. Biased opioid ligands. Molecules 2020; 25(18): 4257. doi: 10.3390/molecules 25184257.
29. Huang H, Bai X, Zhang K, et al. Antinociceptive effects and interaction mechanisms of intrathecal pentazocine and neostigmine in two different pain models in rats. Pain Res Manag 2022; 2022: 4819910. doi: 10.1155/ 2022/4819910.
30. Onasanwo SA, Rotu RA. Antinociceptive and anti-inflammatory potentials of kolaviron: mechanisms of action. J Basic Clin Physiol Pharmacol 2016; 27(4): 363-370.
31. Abdel-Fattah AM, Matsumoto K, Watanabe H. Antinociceptive effects of Nigella sativa oil and its major component, thymoquinone, in mice. Eur J Pharmacol 2000; 400(1): 89-97.
32. Botea MO, Andereggen L, Urman RD, et al. Cannabinoids for acute pain management: approaches and rationale. Curr Pain Headache Rep 2024; 28(7): 681-689.
33. Zubrzycki M, Janecka A, Liebold A, et al. Effects of centrally administered endocannabinoids and opioids on orofacial pain perception in rats. Br J Pharmacol 2017; 174(21): 3780-3789.
34. Anand P, Whiteside G, Fowler CJ, et al. Targeting CB2 receptors and the endocannabinoid system for the treatment of pain. Brain Res Rev 2009; 60(1): 255-266.
35. Xu K, Wu Y, Tian Z, et al. Microglial cannabinoid CB2 receptors in pain modulation. Int J Mol Sci 2023; 24(3): 2348. doi: 10.3390/ijms24032348.
36. Bartolini A, Di Cesare Mannelli L, Ghelardini C. Analgesic and antineuropathic drugs acting through central cholinergic mechanisms. Recent Pat CNS Drug Discov 2011; 6(2):119-140.
37. Jones PG, Dunlop J. Targeting the cholinergic system as a therapeutic strategy for the treatment of pain. Neuropharmacology 2007: 53(2): 197-206.
38. Tamaddonfard E, Hamzeh-Gooshchi N. Effects of subcutaneous and intracerebroventricular injection of physostigmine on the acute corneal nociception in rats. Pharmacol Rep 2010; 62(5): 858-863.
39. el Tahir KE, Ashour MM, al-Harbi MM. The cardiovascular actions of the volatile oil of the black seed (Nigella sativa) in rats: elucidation of the mechanism of action. Gen Pharmacol 1993; 24(5): 1123-1131.
40. Cortes-Altamirano JL, Olmos-Hernandez A, Jaime HB, et al. Review: 5-HT1, 5-HT2, 5-HT3 and 5-HT7 receptors and their role in the modulation of pain response in the central nervous system. Curr Neuropharmacol 2018; 16(2): 210-221.
41. Liao C, Guo J, Rui J, et al. 5-HT3a receptor contributes to neuropathic pain by regulating central sensitization in a rat with brachial plexus avulsion. Physiol Behav 2024; 277: 114503. doi: 10.1016/j.physbeh. 2024.114503.
42. Khalilzadeh E, Saiah GV. The possible mechanisms of analgesia produced by microinjection of morphine into the lateral habenula in the acute model of trigeminal pain in rats. Res Pharm Sci 2017; 12(3): 241-248.
43. Abbas F, Eladl MA, El-Sherbiny M, et al. Celastrol and thymoquinone alleviate aluminum chloride-induced neurotoxicity: behavioral psychomotor performance, neurotransmitter level, oxidative-inflammatory markers, and BDNF expression in rat brain. Biomed Pharmacother 2022; 151: 113072. doi: 10.1016/ j.biopha.2022.113072.
44. Ghoreishi SS, Azizi S, Tamaddonfard E, et al. Thymoquinone attenuates paw incision-induced spontaneous and evoked pain through anti-oxidative and anti-inflammatory mechanisms in rats Vet Res Forum 2025; 16(2): 89-96.
45. Anbarian F, Tamaddonfard E, Erfanparast A, et al. Cerebellar fastigial nucleus histamine and its H2 but not H1 receptors might inhibit acetic acid-induced visceral nociception and improve motor coordination in rats: role of opioid system. Vet Res Forum 2023; 14(10): 549-557.
Veterinary Research Forum
Volume 17, Issue 3
March 2026
Pages 175-181

  • Receive Date 06 January 2025
  • Revise Date 19 March 2025
  • Accept Date 22 April 2025

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