Abstract
MicroRNAs (miRNAs) has been reported to act as key regulators of neuronal function. Increasing evidence has showed that miRNAs exert significant effects in neuropathic pain. We explored the role of miR-30b-5p in neuropathic pain by establishing a rat model of chronic constrictive injury (CCI). The sciatic nerve of CCI rats was used to induce chronic neuropathic pain. The expression and cellular distribution of miR-30b-5p were determined by RT-qPCR and FISH. The mRNA level, protein level, and cellular distribution of CYP24A1 were detected by RT-qPCR, western blot, and immunofluorescence staining assays, respectively. The interaction between miR-30b-5p and CYP24A1 was examined by a luciferase reporter assay. The behavioral effects of miR-30b-5p were assessed after intrathecal administration. Mechanical stimuli and radiant heat were applied to assess mechanical allodynia and thermal hyperalgesia of rats. ELISA was performed to measure the concentration of inflammatory cytokines. MiR-30b-5p expression was significantly downregulated in the spinal cord tissues and of CCI rats. Overexpression of miR-30b-5p attenuated symptoms of neuropathic pain, including mechanical allodynia and thermal hyperalgesia. Additionally, miR-30b-5p overexpression suppressed neuroinflammation by reducing the levels of IL-6, TNF-α and COX2 and elevating the levels of IL-10 in CCI rats. Mechanistically, CYP24A1 was a target of miR-30b-5p, and its expression was negatively regulated by miR-30b-5p. Moreover, CYP24A1 expression was upregulated in CCI rats and knockdown of CYP24A1 attenuated neuropathic pain and neuroinflammation. Furthermore, miR-30b-5p reduced the levels of the Wnt pathway-related genes in CCI rats by downregulating CYP24A1. Rescue assays showed that overexpression of CYP24A1 or activation of Wnt pathway reduced the alleviative effects of miR-30b-5p overexpression on neuropathic pain in CCI rats. Overall, miR-30b-5p inhibits neuropathic pain progression in CCI rats by inhibiting the CYP24A1-Wnt/β-catenin pathway.
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Abbreviations
- miRNAs:
-
MicroRNAs
- CCI:
-
Chronic constrictive injury
- TNF:
-
Tumor necrosis factor
- COX2:
-
Cyclooxygenase-2
References
Agarwal V, Bell GW, Nam JW, Bartel DP (2015) Predicting effective microRNA target sites in mammalian mRNAs. Elife. https://doi.org/10.7554/eLife.05005
Ambros V (2004) The functions of animal microRNAs. Nature 431:350–355. https://doi.org/10.1038/nature02871
Austin PJ, Moalem-Taylor G (2010) The neuro-immune balance in neuropathic pain: involvement of inflammatory immune cells, immune-like glial cells and cytokines. J Neuroimmunol 229:26–50. https://doi.org/10.1016/j.jneuroim.2010.08.013
Backonja MM, Coe CL, Muller DA, Schell K (2008) Altered cytokine levels in the blood and cerebrospinal fluid of chronic pain patients. J Neuroimmunol 195:157–163. https://doi.org/10.1016/j.jneuroim.2008.01.005
Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116:281–297. https://doi.org/10.1016/s0092-8674(04)00045-5
Bennett GJ, Xie YK (1988) A peripheral mononeuropathy in rat that produces disorders of pain sensation like those seen in man. Pain 33:87–107. https://doi.org/10.1016/0304-3959(88)90209-6
Cao S, Wei F, Zhou J, Zhu Z, Li W, Wu M (2020) The synergistic effect between adult weight changes and CYP24A1 polymorphisms is associated with pre- and postmenopausal breast cancer risk. Breast Can Res Treat 179:499–509. https://doi.org/10.1007/s10549-019-05484-6
Carvalho JTG et al (2017) Cholecalciferol decreases inflammation and improves vitamin D regulatory enzymes in lymphocytes in the uremic environment: a randomized controlled pilot trial. PLoS ONE 12:e0179540. https://doi.org/10.1371/journal.pone.0179540
Clark AK, Old EA, Malcangio M (2013) Neuropathic pain and cytokines: current perspectives. J Pain Res 6:803–814. https://doi.org/10.2147/jpr.S53660
Cohen SP, Mao J (2014) Neuropathic pain: mechanisms and their clinical implications. BMJ 348:f7656. https://doi.org/10.1136/bmj.f7656
Costigan M, Scholz J, Woolf CJ (2009) Neuropathic pain: a maladaptive response of the nervous system to damage. Annu Rev Neurosci 32:1–32. https://doi.org/10.1146/annurev.neuro.051508.135531
de Moraes Vieira EB, Garcia JB, da Silva AA, Mualem Araújo RL, Jansen RC (2012) Prevalence, characteristics, and factors associated with chronic pain with and without neuropathic characteristics in São Luís, Brazil. J Pain Symptom Manag 44:239–251. https://doi.org/10.1016/j.jpainsymman.2011.08.014
Derry S, Bell RF, Straube S, Wiffen PJ, Aldington D, Moore RA (2019) Pregabalin for neuropathic pain in adults. Cochrane Database Syst Rev 1:Cd007076. https://doi.org/10.1002/14651858.CD007076.pub3
Dozio E et al (2015) Epicardial adipose tissue inflammation is related to vitamin D deficiency in patients affected by coronary artery disease. Nutr Metab Cardiovasc Dis 25:267–273. https://doi.org/10.1016/j.numecd.2014.08.012
Farrar JT (2010) Advances in clinical research methodology for pain clinical trials. Nat Med 16:1284–1293. https://doi.org/10.1038/nm.2249
Finnerup NB, Sindrup SH, Jensen TS (2010) The evidence for pharmacological treatment of neuropathic pain. Pain 150:573–581. https://doi.org/10.1016/j.pain.2010.06.019
Guo H et al (2020) Nuclear miR-30b-5p suppresses TFEB-mediated lysosomal biogenesis and autophagy. Cell Death Differ. https://doi.org/10.1038/s41418-020-0602-4
Hu P et al (2019) Acidosis enhances the self-renewal and mitochondrial respiration of stem cell-like glioma cells through CYP24A1-mediated reduction of vitamin D. Cell Death Dis 10:25. https://doi.org/10.1038/s41419-018-1242-1
Hu C, Zhao Y, Cui Y, Zhang H, Huang G, Liu Y (2020a) Wnt/beta-catenin signaling contributes to vincristine-induced neuropathic. Pain 69:701–710. https://doi.org/10.33549/physiolres.934314
Hu C, Zhao YT, Cui YB, Zhang HH, Huang GL, Liu Y, Liu YF (2020b) Wnt/beta-catenin signaling contributes to vincristine-induced neuropathic pain. Physiol Res 69:701–710. https://doi.org/10.33549/physiolres.934314
Hung AL, Lim M, Doshi TL (2017) Targeting cytokines for treatment of neuropathic pain. Scand J Pain 17:287–293. https://doi.org/10.1016/j.sjpain.2017.08.002
Ji LJ, Shi J, Lu JM, Huang QM (2018) MiR-150 alleviates neuropathic pain via inhibiting toll-like receptor 5. J Cell Biochem 119:1017–1026. https://doi.org/10.1002/jcb.26269
Kiguchi N, Kobayashi Y, Kishioka S (2012) Chemokines and cytokines in neuroinflammation leading to neuropathic pain. Curr Opin Pharmacol 12:55–61. https://doi.org/10.1016/j.coph.2011.10.007
Kósa JP et al (2013) CYP24A1 inhibition facilitates the anti-tumor effect of vitamin D3 on colorectal cancer cells. World J Gastroenterol 19:2621–2628. https://doi.org/10.3748/wjg.v19.i17.2621
Lewis BP, Burge CB, Bartel DP (2005) Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell 120:15–20. https://doi.org/10.1016/j.cell.2004.12.035
Li L et al (2019) MiR-30b-5p attenuates oxaliplatin-induced peripheral neuropathic pain through the voltage-gated sodium channel Na(v)1.6 in rats. Neuropharmacology 153:111–120. https://doi.org/10.1016/j.neuropharm.2019.04.024
Liu HY, Li YX, Hao YJ, Wang HY, Dai XY, Sun T, Yu JQ (2012) Effects of oxymatrine on the neuropathic pain induced by chronic constriction injury in mice. CNS Neurosci Therap 18:1030–1032. https://doi.org/10.1111/cns.12026
Lopes BC et al (2020) Transcranial direct current stimulation combined with exercise modulates the inflammatory profile and hyperalgesic response in rats subjected to a neuropathic pain model: long-term effects. Brain Stimul 13:774–782. https://doi.org/10.1016/j.brs.2020.02.025
Pandey A et al (2015) Critical role of the miR-200 family in regulating differentiation and proliferation of neurons. J Neurochem 133:640–652. https://doi.org/10.1111/jnc.13089
Pinzone MR et al (2013) LPS and HIV gp120 modulate monocyte/macrophage CYP27B1 and CYP24A1 expression leading to vitamin D consumption and hypovitaminosis D in HIV-infected individuals. Eur Rev Med Pharmacol Sci 17:1938–1950
Sakaki T, Kagawa N, Yamamoto K, Inouye K (2005) Metabolism of vitamin D3 by cytochromes P450. Front Biosci 10:119–134. https://doi.org/10.2741/1514
Sakhaee MH, Sayyadi SAH, Sakhaee N, Sadeghnia HR, Hosseinzadeh H, Nourbakhsh F, Forouzanfar F (2020) Cedrol protects against chronic constriction injury-induced neuropathic pain through inhibiting oxidative stress and inflammation. Metab Brain Dis 35:1119–1126. https://doi.org/10.1007/s11011-020-00581-8
Schneider CA, Rasband WS, Eliceiri KW (2012) NIH Image to ImageJ: 25 years of image analysis. Nat Methods 9:671–675. https://doi.org/10.1038/nmeth.2089
Scholz J, Woolf CJ (2007) The neuropathic pain triad: neurons, immune cells and glia. Nat Neurosci 10:1361–1368. https://doi.org/10.1038/nn1992
Sommer C, Leinders M, Üçeyler N (2018) Inflammation in the pathophysiology of neuropathic pain. Pain 159:595–602. https://doi.org/10.1097/j.pain.0000000000001122
Suh HI, Min J, Choi KH, Kim SW, Kim KS, Jeon SR (2011) Axonal regeneration effects of Wnt3a-secreting fibroblast transplantation in spinal cord-injured rats. Acta Neurochir 153:1003–1010. https://doi.org/10.1007/s00701-011-0945-1
Sun H et al (2018) CYP24A1 inhibition facilitates the antiproliferative effect of 1,25(OH)(2)D(3) through downregulation of the WNT/β-catenin pathway and methylation-mediated regulation of CYP24A1 in Colorectal Cancer Cells. DNA Cell Biol 37:742–749. https://doi.org/10.1089/dna.2017.4058
Uçeyler N, Rogausch JP, Toyka KV, Sommer C (2007) Differential expression of cytokines in painful and painless neuropathies. Neurology 69:42–49. https://doi.org/10.1212/01.wnl.0000265062.92340.a5
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. https://doi.org/10.1016/j.pain.2013.11.013
Wang W, Li R (2020) MiR-216a-5p alleviates chronic constriction injury-induced neuropathic pain in rats by targeting KDM3A and inactivating Wnt/β-catenin signaling pathway. Neurosci Res. https://doi.org/10.1016/j.neures.2020.08.001
Whitehead KJ, Smith CG, Delaney SA, Curnow SJ, Salmon M, Hughes JP, Chessell IP (2010) Dynamic regulation of spinal pro-inflammatory cytokine release in the rat in vivo following peripheral nerve injury. Brain Behav Immun 24:569–576. https://doi.org/10.1016/j.bbi.2009.12.007
Xu Z, Chen Y, Yu J, Yin D, Liu C, Chen X, Zhang D (2015) TCF4 mediates the maintenance of neuropathic pain through Wnt/β-catenin signaling following peripheral nerve injury in rats. J Mol Neurosci 56:397–408. https://doi.org/10.1007/s12031-015-0565-y
Xu S et al (2016) miR-424(322) reverses chemoresistance via T-cell immune response activation by blocking the PD-L1 immune checkpoint. Nat Commun 7:11406. https://doi.org/10.1038/ncomms11406
Yang M et al (2019) Semaphorin 3A contributes to secondary blood-brain barrier damage after traumatic brain injury. Front Cell Neurosci 13:117. https://doi.org/10.3389/fncel.2019.00117
Zhan LY et al (2018) Overexpression of miR-381 relieves neuropathic pain development via targeting HMGB1 and CXCR4. Biomed Pharmacother 107:818–823. https://doi.org/10.1016/j.biopha.2018.08.053
Zhang L, Jia X (2019) Down-regulation of miR-30b-5p protects cardiomyocytes against hypoxia-induced injury by targeting Aven. Cell Mol Biol Lett 24:61. https://doi.org/10.1186/s11658-019-0187-4
Zhang YK, Huang ZJ, Liu S, Liu YP, Song AA, Song XJ (2013) WNT signaling underlies the pathogenesis of neuropathic pain in rodents. J Clin Investig 123:2268–2286. https://doi.org/10.1172/jci65364
Zhang Y, Su Z, Liu HL, Li L, Wei M, Ge DJ, Zhang ZJ (2018) Effects of miR-26a-5p on neuropathic pain development by targeting MAPK6 in in CCI rat models. Biomed Pharmacother 107:644–649. https://doi.org/10.1016/j.biopha.2018.08.005
Zhang W, Suo M, Yu G, Zhang M (2019a) Antinociceptive and anti-inflammatory effects of cryptotanshinone through PI3K/Akt signaling pathway in a rat model of neuropathic pain. Chemico-Biological Interact 305:127–133. https://doi.org/10.1016/j.cbi.2019.03.016
Zhang Y, Liu HL, An LJ, Li L, Wei M, Ge DJ, Su Z (2019b) miR-124–3p attenuates neuropathic pain induced by chronic sciatic nerve injury in rats via targeting EZH2. J Cell Biochem 120:5747–5755. https://doi.org/10.1002/jcb.27861
Zhang Y, Su Z, An LJ, Li L, Wei M, Ge DJ, Liu HL (2019c) miR-98 acts as an inhibitor in chronic constriction injury-induced neuropathic pain via downregulation of high-mobility group AT-hook 2. J Cell Biochem 120:10363–10369. https://doi.org/10.1002/jcb.28320
Zhang Q et al (2020) MiR-30b-5p regulates the lipid metabolism by targeting PPARGC1A in Huh-7 cell line. Lipids Health Dis 19:76. https://doi.org/10.1186/s12944-020-01261-3
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Junfeng Liao and Xiaoyu Lv were the main designers of this study. All authors performed the experiments and analyzed the data. Xiaoyu Lv drafted the manuscript. All authors read and approved the final manuscript.
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Liao, J., Liu, J., Long, G. et al. MiR-30b-5p attenuates neuropathic pain by the CYP24A1-Wnt/β-catenin signaling in CCI rats. Exp Brain Res 240, 263–277 (2022). https://doi.org/10.1007/s00221-021-06253-y
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DOI: https://doi.org/10.1007/s00221-021-06253-y