37: Electroacupuncture alleviates central post stroke pain by modulating miR-21-5p/ TGF-β/Smads pathway

Topic: Basic Science
Subtopic: Pain category (ICD-11), Chronic neuropathic pain, , , , , , , , , , Non-pharmacological interventions

Background and Aims: Background Central post-stroke pain (CPSP) poses a persistent challenge with limited responses to current
pharmacological interventions. Recent evidence implicates miR-21 and its downstream TGF-β/Smads signaling
pathway in pain pathophysiology. This study investigates whether the analgesic effect of electroacupuncture (EA) is
related to miR-21/TGF-β/Smads signaling pathway in CPSP.

Methods: CPSP was induced in rats by collagenase-IV injection into the right thalamic ventral posterolateral nucleus
to produce a focal hemorrhagic stroke lesion. miR-21-5p and Smad7 expressions were manipulated by adenoviral
transfection. Pain thresholds were assessed during 15 days after CPSP induction, followed by brain tissue analysis
using nissl staining, immunofluorescence, immunohistochemistry, and molecular assays. A dual-luciferase reporter
assay was employed to verify the direct binding interaction between miR-21-5p and Smad7.

Results: The results showed EA significantly alleviated CPSP, and meanwhile, downregulated miR-21-5p, upregulated
Smad7, and reduced TGF-β1, p-Smad2, and p-Smad3 expressions, glial hyperactivation and inflammatory factors
(IL-1β, IL-6, TNF-α) in CPSP rats. In addition, miR-21-5p inhibition resulted in analgesic effect, while miR-21-5p
overexpression or Smad7 inhibition counteracted the effect of EA.

Conclusions: These results indicate that the mechanisms underlying EA-induced analgesia involve downregulating
miR-21-5p, consequently inhibiting neuroinflammation by regulating Smad7/TGF-β/Smads signaling pathway,
providing a novel insight for probing the molecular mechanism of analgesic effect of EA.

References: [1] Klit H, Finnerup NB, Jensen TS. Central post-stroke pain: Clinical characteristics, pathophysiology, and management. Lancet Neurol, 2009, 8: 857-868. doi: 10.1016/S1474-4422(09)70176-0.
[2] Yang Y, Yang F, Yang F, et al. Gabapentinoid insensitivity after repeated administration is associated with down-regulation of the α(2)δ-1 subunit in rats with central post-stroke pain hypersensitivity. Neurosci Bull, 2016, 32: 41-50. doi:10.1007/s12264-015-0008-3.
[3] Liu T, Li T, Chen X, et al. Eets/sehi alleviates nociception by blocking the crosslink between endoplasmic reticulum stress and neuroinflammation in a central poststroke pain model. J Neuroinflammation, 2021, 18: 211. doi:10.1186/s12974-021-02255-3.
[4] Tian GH, Tao SS, Chen MT, et al. Electroacupuncture treatment alleviates central poststroke pain by inhibiting brain neuronal apoptosis and aberrant astrocyte activation. Neural Plast, 2016, 2016: 1437148. doi:10.1155/2016/1437148.
[5] Cheng Y, Wu B, Huang J, et al. Research progress on the mechanisms of central post-stroke pain: A review. Cell Mol Neurobiol, 2023, 43: 3083-3098. doi:10.1007/s10571-023-01360-6.
[6] Eisenberg E, McNicol E, Carr DB. Opioids for neuropathic pain. Cochrane Database Syst Rev, 2006, Cd006146. doi:10.1002/14651858.CD006146.
[7] Yang J, Li X, Li C, et al. Comparative efficacy and safety of acupuncture and western medicine for poststroke thalamic pain. Anat Rec (Hoboken), 2023, 306: 3050-3059. doi:10.1002/ar.24902.
[8] Gan JH, Santorelli LA. Acupuncture in post-stroke shoulder pain syndrome with multiple sclerosis: A case study. J Acupunct Meridian Stud, 2021, 14: 27-31. doi:10.51507/j.jams.2021.14.1.27.
[9] Xu XM, Luo H, Rong BB, et al. Nonpharmacological therapies for central poststroke pain: A systematic review. Medicine (Baltimore), 2020, 99: e22611. doi:10.1097/MD.0000000000022611.
[10] Yen HL, Chan W. An east-west approach to the management of central post-stroke pain. Cerebrovasc Dis, 2003, 16: 27-30. doi:10.1159/000070111.
[11] Singer J, Conigliaro A, Spina E, et al. Central poststroke pain: A systematic review. Int J Stroke, 2017, 12: 343-355.doi:10.1177/1747493017701149.
[12] Vas L, Khandagale N, Pai R. Successful management of chronic postsurgical pain following total knee replacement. Pain Med, 2014, 15: 1781-1785. doi:10.1111/pme.12508
[13] Zeboudj L, Sideris-Lampretsas G, Silva R, et al. Silencing mir-21-5p in sensory neurons reverses neuropathic allodynia via activation of tgf-β-related pathway in macrophages. J Clin Invest, 2023, 133. doi:10.1172/JCI164472.
[14] Simeoli R, Montague K, Jones HR, et al. Exosomal cargo including microrna regulates sensory neuron to macrophage communication after nerve trauma. Nat Commun, 2017, 8: 1778. doi:10.1038/s41467-017-01841-5.
[15] Karl-Schöller F, Kunz M, Kreß L, et al. A translational study: Involvement of mir-21-5p in development and maintenance of neuropathic pain via immune-related targets ccl5 and ywhae. Exp Neurol, 2022, 347: 113915. doi:10.1016/j.expneurol.2021.113915.
[16] Men Y, Yelick J, Jin S, et al. Exosome reporter mice reveal the involvement of exosomes in mediating neuron to astroglia communication in the cns. Nat Commun, 2019, 10: 4136. doi:10.1038/s41467-019-11534-w.
[17] Reyes-Long S, Cortés-Altamirano JL, Bandala C, et al. Role of the micrornas in the pathogenic mechanism of painful symptoms in long covid: Systematic review. Int J Mol Sci, 2023, 24(4):3574. doi:10.3390/ijms24043574.
[18] Li W, Yu X, Chen X, et al. Hbv induces liver fibrosis via the tgf-β1/mir-21-5p pathway. Exp Ther Med, 2021, 21: 169. doi:10.3892/etm.2020.9600.
[19] Zhang X, Chen Y, Yang M, et al. Mir-21-5p actions at the smad7 gene during pig ovarian granulosa cell apoptosis. Anim Reprod Sci, 2020, 223: 106645. doi:10.1016/j.anireprosci.2020.106645.
[20] Wu XD, Kang L, Tian J, et al. Exosomes derived from magnetically actuated bone mesenchymal stem cells promote tendon-bone healing through the mir-21-5p/smad7 pathway. Mater Today Bio, 2022, 15: 100319. doi:10.1016/j.mtbio.2022.100319
[21] Tang J, Li X, Cheng T, et al. Mir-21-5p/smad7 axis promotes the progress of lung cancer. Thorac Cancer, 2021, 12: 2307-2313. doi:10.1111/1759-7714.14060.
[22] Dixit AK, Sarver AE, Yuan Z, et al. Comprehensive analysis of microrna signature of mouse pancreatic acini: Overexpression of mir-21-3p in acute pancreatitis. Am J Physiol Gastrointest Liver Physiol, 2016, 311: G974-g980. doi:10.1152/ajpgi.00191.2016.
[23] Wasserman JK, Koeberle PD. Development and characterization of a hemorrhagic rat model of central post-stroke pain. Neuroscience, 2009, 161: 173-183. doi:10.1016/j.neuroscience.2009.03.042.
[24] Ma G, Chen F, Liu Y, et al. Nur77 ameliorates age-related renal tubulointerstitial fibrosis by suppressing the tgf-β/smads signaling pathway. Faseb j, 2022, 36: e22124. doi:10.1096/fj.202101332R.
[25] Zhan J, Liu M, Pan L, et al. Oxidative stress and tgf-β1/smads signaling are involved in rosa roxburghii fruit extract alleviating renal fibrosis. Evid Based Complement Alternat Med, 2019, 2019: 4946580. doi:10.1155/2019/4946580.
[26] Urits I, Gress K, Charipova K, et al. Diagnosis, treatment, and management of dejerine-roussy syndrome: A comprehensive review. Curr Pain Headache Rep, 2020, 24: 48. doi:10.1007/s11916-020-00887-3.
[27] Zheng L, Li XY, Huang FZ, et al. Effect of electroacupuncture on relieving central post-stroke pain by inhibiting autophagy in the hippocampus. Brain Res, 2020, 1733: 146680. doi:10.1016/j.brainres.2020.146680.
[28] Huang T, Li Y, Hu W, et al. Dexmedetomidine attenuates haemorrhage-induced thalamic pain by inhibiting the tlr4/nf-κb/erk1/2 pathway in mice. Inflammopharmacology, 2021, 29: 1751-1760. doi:10.1007/s10787-021-00877-w.
[29] Yang F, Luo WJ, Sun W, et al. Sdf1-cxcr4 signaling maintains central post-stroke pain through mediation of glial-neuronal interactions. Front Mol Neurosci, 2017, 10: 226. doi:10.3389/fnmol.2017.00226.
[30] Ma Y, Wang J, Wang Y, et al. The biphasic function of microglia in ischemic stroke. Prog Neurobiol, 2017, 157: 247-272. doi:10.1016/j.pneurobio.2016.01.005.
[31] Dworsky-Fried Z, Kerr BJ, Taylor AMW. Microbes, microglia, and pain. Neurobiol Pain, 2020, 7: 100045. doi:10.1016/j.ynpai.2020.100045.
[32] Hiraga SI, Itokazu T, Hoshiko M, et al. Microglial depletion under thalamic hemorrhage ameliorates mechanical allodynia and suppresses aberrant axonal sprouting. JCI Insight, 2020, 5(3): e131801. doi:10.1172/jci.insight.131801.
[33] Zhang R, Lao L, Ren K, et al. Mechanisms of acupuncture-electroacupuncture on persistent pain. Anesthesiology, 2014, 120: 482-503. doi:10.1097/ALN.000000000000010
[34] Xu R, Wang J, Nie H, et al. Genome-wide expression profiling by rna-sequencing in spinal cord dorsal horn of a rat chronic postsurgical pain model to explore potential mechanisms involved in chronic pain. J Pain Res, 2022, 15: 985-1001. doi:10.2147/JPR.S358942
[35] Bai X, Bian Z. Microrna-21 is a versatile regulator and potential treatment target in central nervous system disorders. Front Mol Neurosci, 2022, 15: 842288. doi:10.3389/fnmol.2022.842288
[36] Gada Y, Pandey A, Jadhav N, et al. New vistas in microrna regulatory interactome in neuropathic pain. Front Pharmacol, 2021, 12: 778014. doi:10.3389/fphar.2021.778014.
[37] Zhong L, Xiao W, Wang F, et al. Mir-21-5p inhibits neuropathic pain development via directly targeting c-c motif ligand 1 and tissue inhibitor of metalloproteinase-3. J Cell Biochem, 2019, 120: 16614-16623. doi:10.1002/jcb.28920.
[38] Liu L, Liu L, Liu R, et al. Exosomal mir-21-5p derived from multiple myeloma cells promote renal epithelial-mesenchymal transition through targeting tgf-β/smad7 signalling pathway. Clin Exp Pharmacol Physiol, 2023, 50: 711-718. doi:10.1111/1440-1681.13768.
[39] Fang Y, Zhang Q, Lv C, et al. Mitochondrial fusion induced by transforming growth factor-β1 serves as a switch that governs the metabolic reprogramming during differentiation of regulatory t cells. Redox Biol, 2023, 62: 102709. doi:10.1016/j.redox.2023.102709.
[40] Luo L, Gao Y, Yang C, et al. Halofuginone attenuates intervertebral discs degeneration by suppressing collagen i production and inactivating tgfβ and nf-кb pathway. Biomed Pharmacother, 2018, 101: 745-753. doi:10.1016/j.biopha.2018.01.100.
[41] Chen L, Yang T, Lu DW, et al. Central role of dysregulation of tgf-β/smad in ckd progression and potential targets of its treatment. Biomed Pharmacother, 2018, 101: 670-681. doi:10.1016/j.biopha.2018.02.090.
[42] Yan X, Chen YG. Smad7: Not only a regulator, but also a cross-talk mediator of tgf-β signalling. Biochem J, 2011, 434: 1-10. doi:10.1042/BJ20101827.

Presenting Author: None
Submitting Author: Zhihao Shang

Senior Author: Zhihao Shang

IASP 2026 Global Year theme of Neuropathic Pain: No