Intrathecal injection of ozone alleviates CCI‑induced neuropathic pain via the GluR6‑NF‑κB/p65 signalling pathway in rats

Zhang,Weiguang; Wang,Fei; Zhang,Licai; Sun,Tao; Fu,Zhijian

doi:10.3892/mmr.2021.11870

Intrathecal injection of ozone alleviates CCI‑induced neuropathic pain via the GluR6‑NF‑κB/p65 signalling pathway in rats

Authors:
- Weiguang Zhang
- Fei Wang
- Licai Zhang
- Tao Sun
- Zhijian Fu
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Affiliations: Department of Pain Management, Shandong Provincial Hospital, Cheeloo College of Medicine, Shandong University, Jinan, Shandong 250021, P.R. China, Department of Pain Management, Shandong Linyi People's Hospital, Linyi, Shandong 276000, P.R. China, Key Laboratory of Anaesthesiology of Jiangsu Province and Key Laboratory of Anaesthesia and Analgesia Application Technology of Jiangsu Province, Xuzhou Medical University, Xuzhou, Jiangsu 221004, P.R. China
Published online on: January 26, 2021 https://doi.org/10.3892/mmr.2021.11870
Article Number: 231

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Abstract

Ozone is widely used to relieve chronic pain clinically, but the precise mechanisms governing its action have yet to be elucidated. The present study aimed to investigate the mechanisms underlying the pain‑alleviating effect of ozone in the chronic constriction injury (CCI) model of sciatic nerve in rats. Pain behaviours of rats were assessed by mechanical allodynia and thermal hyperalgesia. The expression of spinal glutamate receptor 6 (GluR6) and NF‑κB/p65 was detected by western blotting and reverse transcription‑quantitative PCR. Meanwhile, the expression of spinal IL‑1β, IL‑6 and TNF‑α was detected by ELISA. GluR6 short interfering (si)RNAs were used intrathecally immediately following CCI once per day. Ozone (10, 20 or 30 µg/ml) or oxygen was injected intrathecally on day 7 after CCI. The expression level of spinal GluR6 increased on day 3 and reached a peak on day 7 after CCI. The expression level of spinal IL‑1β, IL‑6, TNF‑α and NF‑κB/p65 also increased on day 7 after CCI. In addition, pre‑intrathecal injection of GluR6 siRNAs inhibited pain behaviours and suppressed the expression of spinal GluR6, IL‑1β, IL‑6, TNF‑α and NF‑κB/p65 in CCI rats on day 7. Intrathecal injection of ozone was also observed to inhibit pain behaviours and suppress the expression of spinal GluR6, IL‑1β, IL‑6, TNF‑α and NF‑κB/p65 in CCI rats on day 7. The present study suggested that GluR6 served a pivotal role in neuropathic pain and that intrathecal injection of ozone may alleviate neuropathic pain via the GluR6‑NF‑κB/p65 signalling pathway.

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1	Liu JC, Xue DF, Wang XQ, Ai DB and Qin PJ: MiR-101 relates to chronic peripheral neuropathic pain through targeting KPNB1 and regulating NF-κB signaling. Kaohsiung J Med Sci. 35:139–145. 2019. View Article : Google Scholar : PubMed/NCBI
2	Baron R, Binder A and Wasner G: Neuropathic pain: Diagnosis, pathophysiological mechanisms, and treatment. Lancet Neurol. 9:807–819. 2010. View Article : Google Scholar : PubMed/NCBI
3	Zimmermann M: Pathobiology of neuropathic pain. Eur J Pharmacol. 429:23–37. 2001. View Article : Google Scholar : PubMed/NCBI
4	Ma X, Wang H, Song T, Wang W and Zhang Z: lncRNA MALAT1 contributes to neuropathic pain development through regulating miR-129-5p/HMGB1 axis in a rat model of chronic constriction injury. Int J Neurosci. 130:1215–1224. 2020. View Article : Google Scholar : PubMed/NCBI
5	Baron R: Mechanisms of disease: Neuropathic pain-a clinical perspective. Nat Clin Pract Neurol. 2:95–106. 2006. View Article : Google Scholar : PubMed/NCBI
6	Pu S, Li S, Xu Y, Wu J, Lv Y and Du D: Role of receptor-interacting protein 1/receptor-interacting protein 3 in inflammation and necrosis following chronic constriction injury of the sciatic nerve. Neuroreport. 29:1373–1378. 2018. View Article : Google Scholar : PubMed/NCBI
7	Xu L, Liu Y, Sun Y, Li H, Mi W and Jiang Y: Analgesic effects of TLR4/NF-κB signaling pathway inhibition on chronic neuropathic pain in rats following chronic constriction injury of the sciatic nerve. Biomed Pharmacother. 107:526–533. 2018. View Article : Google Scholar : PubMed/NCBI
8	Al Habashneh R, Alsalman W and Khader Y: Ozone as an adjunct to conventional nonsurgical therapy in chronic periodontitis: A randomized controlled clinical trial. J Periodontal Res. 50:37–43. 2015. View Article : Google Scholar : PubMed/NCBI
9	Magalhaes FN, Dotta L, Sasse A, Teixera MJ and Fonoff ET: Ozone therapy as a treatment for low back pain secondary to herniated disc: A systematic review and meta-analysis of randomized controlled trials. Pain Physician. 15:E115–E129. 2012.PubMed/NCBI
10	Biazzo A, Corriero AS and Confalonieri N: Intramuscular oxygen-ozone therapy in the treatment of low back pain. Acta Biomed. 89:41–46. 2018.PubMed/NCBI
11	Costa T, Linhares D, Ribeiro da Silva M and Neves N: Ozone therapy for low back pain, A systematic review. Acta Reumatol Port. 43:172–181. 2018.PubMed/NCBI
12	Crockett MT, Moynagh M, Long N, Kilcoyne A, Dicker P, Synnott K and Eustace SJ: Ozone-augmented percutaneous discectomy: A novel treatment option for refractory discogenic sciatica. Clin Radiol. 69:1280–1286. 2014. View Article : Google Scholar : PubMed/NCBI
13	Ozcan S, Muz A, Yildiz Altun A and Onal SA: Intradiscal ozone therapy for lumbar disc herniation. Cell Mol Biol. 64:52–55. 2018. View Article : Google Scholar : PubMed/NCBI
14	Magalhães FN, Soares SC, Torres JM, Ungaretti A, Cacciacarro MF, Teixeira MJ and Fonoff ET: Effects of ozone applied by spinal endoscopy in patients with chronic pain related to failed back surgery syndrome: A pilot study. Neuropsychiatr Dis Treat. 9:1759–1766. 2013.PubMed/NCBI
15	Barbosa DC, Ângelos JSD, Macena GMJ, Magalhães FNO and Fonoff ET: Effects of ozone on the pain and disability in patients with failed back surgery syndrome. Rev Assoc Med Bras. 63:355–360. 2017. View Article : Google Scholar : PubMed/NCBI
16	Mauro RD, Cantarella G, Bernardini R, Rosa MD, Barbagallo I, Distefano A, Longhitano L, Vicario N, Nicolosi D, Lazzarino G, et al: The biochemical and pharmacological properties of ozone: The smell of protection in acute and chronic diseases. Int J Mol Sci. 20:6342019. View Article : Google Scholar
17	Fuccio C, Luongo C, Capodanno P, Giordano C, Scafuro MA, Siniscalco D, Lettieri B, Rossi F, Maione S and Berrino L: A single subcutaneous injection of ozone prevents allodynia and decreases the over-expression of proinflammatory caspases in the orbito-frontal cortex of neuropathic mice. Eur J Pharmacol. 603:42–49. 2009. View Article : Google Scholar : PubMed/NCBI
18	Holz O, Tal-Singer R, Kanniess F, Simpson KJ, Gibson A, Vessey RS, Janicki S, Magnussen H, Jörres RA and Richter K: Validation of the human ozone challenge model as a tool for assessing anti-inflammatory drugs in early development. J Clin Pharmacol. 45:498–503. 2005. View Article : Google Scholar : PubMed/NCBI
19	Zamora ZB, Borrego A, Lopez OY, Delgado R, González R, Menéndez S, Hernández F and Schulz S: Effects of ozone oxidative preconditioning on TNF-alpha release and antioxidant-prooxidant intracellular balance in mice during endotoxic shock. Mediators Inflamm. 2005:16–22. 2005. View Article : Google Scholar : PubMed/NCBI
20	Lu Y, Sun YN, Wu X, Sun Q, Liu FY, Xing GG and Wan Y: Role of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptor subunit GluR1 in spinal dorsal horn in inflammatory nociception and neuropathic nociception in rat. Brain Res. 1200:19–26. 2008. View Article : Google Scholar : PubMed/NCBI
21	Marshall J, Blair LA and Singer JD: BTB-Kelch proteins and ubiquitination of kainate receptors. Adv Exp Med Biol. 717:115–125. 2011. View Article : Google Scholar : PubMed/NCBI
22	Nasu-Nishimura Y, Jaffe H, Isaac JT and Roche KW: Differential regulation of kainate receptor trafficking by phosphorylation of distinct sites on GluR6. J Biol Chem. 285:2847–2856. 2010. View Article : Google Scholar : PubMed/NCBI
23	Wu LJ, Ko SW and Zhuo M: Kainate receptors and pain: From dorsal root ganglion to the anterior cingulate cortex. Curr Pharm Des. 13:1597–1605. 2007. View Article : Google Scholar : PubMed/NCBI
24	Zhang WG, Zhang LC, Peng ZD and Zeng YM: Intrathecal injection of GluR6 antisense oligodeoxynucleotides alleviates acute inflammatory pain of rectum in rats. Neurosci Bull. 25:319–323. 2009. View Article : Google Scholar : PubMed/NCBI
25	Luo JG, Zhao XL, Xu WC, Zhao XJ, Wang JN, Lin XW, Sun T and Fu ZJ: Activation of spinal NF-κB/p65 contributes to peripheral inflammation and hyperalgesia in rat adjuvant-induced arthritis. Arthritis Rheumatol. 66:896–906. 2014. View Article : Google Scholar : PubMed/NCBI
26	Ni H, Wang Y, An K, Liu Q, Xu L, Zhu C, Deng H, He Q, Wang T, Xu M, et al: Crosstalk between NFκB-dependent astrocytic CXCL1 and neuron CXCR2 plays a role in descending pain facilitation. J Neuroinflammation. 16:12019. View Article : Google Scholar : PubMed/NCBI
27	Pei WM, Zou Y, Wang WT, Wei L, Zhao Y and Li L: Tizanidine exerts anti-nociceptive effects in spared nerve injury model of neuropathic pain through inhibition of TLR4/NF-κB pathway. Int J Mol Med. 42:3209–3219. 2018.PubMed/NCBI
28	Sun W, Zhang L and Li R: Overexpression of miR-206 ameliorates chronic constriction injury-induced neuropathic pain in rats via the MEK/ERK pathway by targeting brain-derived neurotrophic factor. Neurosci Lett. 646:68–74. 2017. View Article : Google Scholar : PubMed/NCBI
29	Cao JL, He JH, Ding HL and Zeng YM: Activation of the spinal ERK signaling pathway contributes naloxone-precipitated withdrawal in morphine-dependent rats. Pain. 118:336–349. 2005. View Article : Google Scholar : PubMed/NCBI
30	Sun T, Luo J, Jia M, Li H, Li K and Fu Z: Small interfering RNA-mediated knockdown of NF-κBp65 attenuates neuropathic pain following peripheral nerve injury in rats. Eur J Pharmacol. 682:79–85. 2012. View Article : Google Scholar : PubMed/NCBI
31	Wang J, Wu M, Lin X, Li Y and Fu Z: Low-concentration oxygen/ozone treatment attenuated radiculitis and mechanical allodynia via PDE2A-cAMP/cGMP-NF-κB/p65 signaling in chronic radiculitis rats. Pain Res Manag. 13:51928142018.
32	Liu S, Liu YP, Song WB and Song XJ: EphrinB-EphB receptor signaling contributes to bone cancer pain via Toll-like receptor and proinflammatory cytokines in rat spinal cord. Pain. 154:2823–2835. 2013. View Article : Google Scholar : PubMed/NCBI
33	Wei XE, Zhang FY, Wang K, Zhang QX and Rong LQ: Fasudil hydrochloride protects neurons in rat hippocampal CA1 region through inhibiting GluR6-MLK3-JNKs signal pathway. Cell Biochem Biophys. 70:415–421. 2014. View Article : Google Scholar : PubMed/NCBI
34	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
35	Giurazza F, Guarnieri G, Murphy KJ and Muto M: Intradiscal O₂ O₃: Rationale, injection technique, short- and long-term outcomes for the treatment of low back pain due to disc herniation. Can Assoc Radiol J. 68:171–177. 2017. View Article : Google Scholar : PubMed/NCBI
36	Raeissadat SA, Tabibian E, Rayegani SM, Dehgolan SR and Ghazani AB: An investigation into the efficacy of intra-articular ozone (O₂-O₃) injection in patients with knee osteoarthritis: A systematic review and meta-analysis. J Pain Res. 11:2537–2550. 2018. View Article : Google Scholar : PubMed/NCBI
37	Bocci V: Ozone as Janus: This controversial gas can be either toxic or medically useful. Mediators Inflamm. 13:3–11. 2004. View Article : Google Scholar : PubMed/NCBI
38	Zhou NB, Fu ZJ and Sun T: Effects of different concentrations of oxygen-ozone on rats' astrocytes in vitro. Neurosci Lett. 441:178–182. 2008. View Article : Google Scholar : PubMed/NCBI
39	Li Y, Lin X, Zhao X, Xie J, Wang J, Sun T and Fu Z: Ozone (O3) elicits neurotoxicity in spinal cord neurons (SCNs) by inducing ER Ca(2+) release and activating the CaMKII/MAPK signaling pathway. Toxicol Appl Pharmacol. 280:493–501. 2014. View Article : Google Scholar : PubMed/NCBI
40	Wu MY, Xing CY, Wang JN, Li Y, Lin XW and Fu ZJ: Therapeutic dosage of ozone inhibits autophagy and apoptosis of nerve roots in a chemically induced radiculoneuritis rat model. Eur Rev Med Pharmacol Sci. 22:1787–1797. 2018.PubMed/NCBI
41	Wei A, Feng H, Jia XM, Tang H, Liao YY and Li BR: Ozone therapy ameliorates inflammation and endometrial injury in rats with pelvic inflammatory disease. Biomed Pharmacother. 107:1418–1425. 2018. View Article : Google Scholar : PubMed/NCBI
42	Re L, Sanchez GM and Mawsouf N: Clinical evidence of ozone interaction with pain mediators. Saudi Med J. 31:1363–1367. 2010.PubMed/NCBI
43	Zhang J, Yan H, Wu YP, Li C and Zhang GY: Activation of GluR6-containing kainate receptors induces ubiquitin-dependent Bcl-2 degradation via denitrosylation in the rat hippocampus after kainate treatment. J Biol Chem. 286:7669–7680. 2011. View Article : Google Scholar : PubMed/NCBI
44	Nishimura YN, Jaffe H, Isaac JT and Roche KW: Differential regulation of kainate receptor trafficking by phosphorylation of distinct sites on GluR6. J Biol Chem. 285:2847–2856. 2010. View Article : Google Scholar : PubMed/NCBI
45	Pei DS, Guan QH, Sun YF, Zhang QX, Xu TL and Zhang GY: Neuroprotective effects of GuR6 antisense oligodeoxynucleotides on transient brain ischemia/reperfusion induced neuronal death in rat hippocampal CA1 region. J Neurosci Res. 82:642–649. 2005. View Article : Google Scholar : PubMed/NCBI
46	Lu CR, Willcockson HH, Phend KD, Lucifora S, Darstein M, Valtschanoff JG and Rustioni A: Ionotropic glutamate receptors are expressed in GABAergic terminals in the rat superficial dorsal horn. J Comp Neurol. 486:169–178. 2005. View Article : Google Scholar : PubMed/NCBI
47	Liu C, Zhang F, Liu H and Wei F: NF-κB mediated CX3CL1 activation in the dorsal root ganglion contributes to the maintenance of neuropathic pain induced in adult male Sprague Dawley rats1. Acta Cir Bras. 33:619–628. 2018. View Article : Google Scholar : PubMed/NCBI
48	Li Y, Yang Y, Guo J, Guo X, Feng Z and Zhao X: Spinal NF-κB upregulation contributes to hyperalgesia in a rat model of advanced osteoarthritis. Mol Pain. 16:17448069209056912020. View Article : Google Scholar : PubMed/NCBI
49	Chou CW, Wong GT, Lim G, Wang S, Irwin MG and Mao J: Spatiotemporal pattern of concurrent spinal and supraspinal NF-κB expression after peripheral nerve injury. J Pain. 12:13–21. 2011. View Article : Google Scholar : PubMed/NCBI
50	Lu L, Pan C, Chen L, Hu L, Wang C, Han Y, Cheng Z, Yang Y and Liu W: AMPK activation by peri-sciatic nerve administration of ozone attenuates CCI induced neuropathic pain in rats. J Mol Cell Biol. 9:132–143. 2017. View Article : Google Scholar : PubMed/NCBI