{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T03:58:09Z","timestamp":1791863889383,"version":"4.3.4"},"reference-count":57,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2020,3,27]],"date-time":"2020-03-27T00:00:00Z","timestamp":1585267200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003329","name":"Ministerio de Econom\u00eda y Competitividad","doi-asserted-by":"publisher","award":["SAF 2014-59701"],"award-info":[{"award-number":["SAF 2014-59701"]}],"id":[{"id":"10.13039\/501100003329","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Peripheral nerve injury (PNI) leads to the loss of motor, sensory, and autonomic functions, and often triggers neuropathic pain. During the last years, many efforts have focused on finding new therapies to increase axonal regeneration or to alleviate painful conditions. Still only a few of them have targeted both phenomena. Incipient or aberrant sensory axon regeneration is related to abnormal unpleasant sensations, such as hyperalgesia or allodynia. We recently have discovered NeuroHeal, a combination of two repurposed drugs; Acamprosate and Ribavirin. NeuroHeal is a neuroprotective agent that also enhances motor axon regeneration after PNI. In this work, we investigated its effect on sensory fiber regeneration and PNI-induced painful sensations in a rat model of spare nerve injury and nerve crush. The follow up of the animals showed that NeuroHeal treatment reduced the signs of neuropathic pain in both models. Besides, the treatment favored sensory axon regeneration, as observed in dorsal root ganglion explants. Mechanistically, the effects observed in vivo may improve the resolution of cell-protective autophagy. Additionally, NeuroHeal treatment modulated the P2X4-BDNF-KCC2 axis, which is an essential driver of neuropathic pain. These data open a new therapeutic avenue based on autophagic modulation to foster endogenous regenerative mechanisms and reduce the appearance of neuropathic pain in PNI.<\/jats:p>","DOI":"10.3390\/cells9040808","type":"journal-article","created":{"date-parts":[[2020,4,1]],"date-time":"2020-04-01T05:52:30Z","timestamp":1585720350000},"page":"808","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["NeuroHeal Treatment Alleviates Neuropathic Pain and Enhances Sensory Axon Regeneration"],"prefix":"10.3390","volume":"9","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3368-9302","authenticated-orcid":false,"given":"David","family":"Romeo-Guitart","sequence":"first","affiliation":[{"name":"Institut de Neuroci\u00e8ncies (INc) and Department of Cell Biology, Physiology and Immunology, Universitat Aut\u00f2noma de Barcelona (UAB), 08193 Bellaterra, Barcelona, Spain"},{"name":"Institut Necker Enfants-Malades (INEM), INSERM U1151, Laboratory \"Hormonal Regulation of Brain Development and Functions\"\u2014Team 8, Universit\u00e9 Paris Descartes, Sorbonne Paris Cit\u00e9, 75015 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2273-6321","authenticated-orcid":false,"given":"Caty","family":"Casas","sequence":"additional","affiliation":[{"name":"Institut de Neuroci\u00e8ncies (INc) and Department of Cell Biology, Physiology and Immunology, Universitat Aut\u00f2noma de Barcelona (UAB), 08193 Bellaterra, Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,3,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Grinsell, D., and Keating, C.P. (2014). Peripheral Nerve Reconstruction after Injury: A Review of Clinical and Experimental Therapies. Biomed Res. Int., 2014.","DOI":"10.1155\/2014\/698256"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"116","DOI":"10.1097\/00005373-199807000-00025","article-title":"Analysis of upper and lower extremity peripheral nerve injuries in a population of patients with multiple injuries","volume":"45","author":"Noble","year":"1998","journal-title":"J. Trauma"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"826","DOI":"10.1016\/j.expneurol.2014.09.006","article-title":"Improving peripheral nerve regeneration: From molecular mechanisms to potential therapeutic targets","volume":"261","author":"Chan","year":"2014","journal-title":"Exp. Neurol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"17002","DOI":"10.1038\/nrdp.2017.2","article-title":"Neuropathic pain","volume":"3","author":"Colloca","year":"2017","journal-title":"Nat. Rev. Dis. Prim."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1016\/j.cell.2009.09.028","article-title":"Cellular and Molecular Mechanisms of Pain","volume":"139","author":"Basbaum","year":"2009","journal-title":"Cell"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1","DOI":"10.4196\/kjpp.2019.23.1.1","article-title":"An integrated review on new targets in the treatment of neuropathic pain","volume":"23","author":"Khangura","year":"2019","journal-title":"Korean J. Physiol. Pharmacol. Off. J. Korean Physiol. Soc. Korean Soc. Pharmacol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"282","DOI":"10.1016\/j.brainresrev.2011.03.003","article-title":"Cellular and molecular insights into neuropathy-induced pain hypersensitivity for mechanism-based treatment approaches","volume":"67","author":"Berger","year":"2011","journal-title":"Brain Res. Rev."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1080\/14728222.2018.1420781","article-title":"Cellular and molecular mechanisms driving neuropathic pain: Recent advancements and challenges","volume":"22","author":"Fernandes","year":"2018","journal-title":"Expert Opin. Ther. Targets"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1093\/bja\/aen107","article-title":"Neuropathic pain: Emerging treatments","volume":"101","author":"Dray","year":"2008","journal-title":"Br. J. Anaesth."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1002\/jcp.21280","article-title":"Ionic channels and neuropathic pain: Physiopathology and applications","volume":"215","author":"Aurilio","year":"2008","journal-title":"J. Cell. Physiol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1016\/j.lfs.2017.08.016","article-title":"Leptin is essential for microglial activation and neuropathic pain after preganglionic cervical root avulsion","volume":"187","author":"Chang","year":"2017","journal-title":"Life Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"291","DOI":"10.3389\/fphar.2017.00291","article-title":"P2X4 Receptor Function in the Nervous System and Current Breakthroughs in Pharmacology","volume":"8","author":"Stokes","year":"2017","journal-title":"Front. Pharmacol."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Jensen, M.P., and Brownstone, R.M. (2018). Mechanisms of Spinal Cord Stimulation for the Treatment of Pain: Still in the Dark after 50 Years. Eur. J. Pain.","DOI":"10.1002\/ejp.1336"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"495","DOI":"10.2174\/1570159X14666160330105132","article-title":"Neuroprotective Effects of Exercise Treatments After Injury: The Dual Role of Neurotrophic Factors","volume":"15","author":"Cobianchi","year":"2017","journal-title":"Curr. Neuropharmacol."},{"key":"ref_15","unstructured":"Brooks, K.G., and Kessler, T.L. (2017). Treatments for neuropathic pain. Clin. Pharm., 1\u201322."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"531","DOI":"10.1038\/s41419-018-0553-6","article-title":"SIRT1 activation with neuroheal is neuroprotective but SIRT2 inhibition with AK7 is detrimental for disconnected motoneurons","volume":"9","author":"Sima","year":"2018","journal-title":"Cell Death Dis."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1879","DOI":"10.1038\/s41598-018-19767-3","article-title":"Neuroprotective Drug for Nerve Trauma Revealed Using Artificial Intelligence","volume":"8","author":"Valls","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.jneumeth.2014.08.005","article-title":"Neurite-J: An Image-J plug-in for axonal growth analysis in organotypic cultures","volume":"236","author":"Santos","year":"2014","journal-title":"J. Neurosci. Methods"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1016\/j.expneurol.2009.01.014","article-title":"Nociceptive responses and spinal plastic changes of afferent C-fibers in three neuropathic pain models induced by sciatic nerve injury in the rat","volume":"217","author":"Vivo","year":"2009","journal-title":"Exp. Neurol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.expneurol.2014.02.008","article-title":"Assessment of sensory thresholds and nociceptive fiber growth after sciatic nerve injury reveals the differential contribution of collateral reinnervation and nerve regeneration to neuropathic pain","volume":"255","author":"Cobianchi","year":"2014","journal-title":"Exp. Neurol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"12028","DOI":"10.1038\/s41598-017-11086-3","article-title":"Boosted Regeneration and Reduced Denervated Muscle Atrophy by NeuroHeal in a Pre-clinical Model of Lumbar Root Avulsion with Delayed Reimplantation","volume":"7","author":"Navarro","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Romeo-Guitart, D., Leiva-Rodr\u00edguez, T., For\u00e9s, J., and Casas, C. (2019). Improved Motor Nerve Regeneration by SIRT1\/Hif1a-Mediated Autophagy. Cells, 8.","DOI":"10.3390\/cells8111354"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1186\/s13041-018-0354-y","article-title":"TLR4-mediated autophagic impairment contributes to neuropathic pain in chronic constriction injury mice","volume":"11","author":"Piao","year":"2018","journal-title":"Mol. Brain"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1186\/1744-8069-7-83","article-title":"Autophagy impairment in a mouse model of neuropathic pain","volume":"7","author":"Berliocchi","year":"2011","journal-title":"Mol. Pain"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"229","DOI":"10.3344\/kjp.2015.28.4.229","article-title":"Autophagy: Can It be a New Experimental Research Method of Neuropathic Pain?","volume":"28","author":"Jung","year":"2015","journal-title":"Korean J. Pain"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Romeo-Guitart, D., Marcos-DeJuana, C., Navarro, X., and Casas, C. (2020). Novel Neuroprotective Therapy with NeuroHeal by Autophagy Induction for Damaged Neonatal Motoneurons. Theranostics, In press.","DOI":"10.7150\/thno.43765"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1080\/15548627.2015.1100356","article-title":"Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)","volume":"12","author":"Klionsky","year":"2016","journal-title":"Autophagy"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"613","DOI":"10.1097\/j.pain.0000000000000410","article-title":"The \u03b15 subunit containing GABAA receptors contribute to chronic pain","volume":"157","author":"Corleto","year":"2016","journal-title":"Pain"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"183","DOI":"10.1038\/nn.3295","article-title":"Morphine hyperalgesia gated through microglia-mediated disruption of neuronal Cl(-) homeostasis","volume":"16","author":"Ferrini","year":"2013","journal-title":"Nat. Neurosci."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1017","DOI":"10.1038\/nature04223","article-title":"BDNF from microglia causes the shift in neuronal anion gradient underlying neuropathic pain","volume":"438","author":"Coull","year":"2005","journal-title":"Nature"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/j.neuroscience.2014.05.044","article-title":"BDNF: No gain without pain?","volume":"283","author":"Smith","year":"2014","journal-title":"Neuroscience"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Hu, B., Senkler, C., Yang, A., Soto, F., and Liang, B.T. (2002). P2X 4 Receptor Is a Glycosylated Cardiac Receptor Mediating a Positive Inotropic Response to ATP *. 277, 15752\u201315757.","DOI":"10.1074\/jbc.M112097200"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1122","DOI":"10.4103\/1673-5374.251187","article-title":"Network-centric medicine for peripheral nerve injury: Treating the whole to boost endogenous mechanisms of neuroprotection and regeneration","volume":"14","author":"Casas","year":"2019","journal-title":"Neural Regen. Res."},{"key":"ref_34","first-page":"361","article-title":"Autophagy of neuron axon during regeneration of rat sciatic nerves","volume":"24","author":"Piao","year":"2004","journal-title":"Di Yi Jun Yi Da Xue Xue Bao"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"11324","DOI":"10.1073\/pnas.1611282113","article-title":"Autophagy induction stabilizes microtubules and promotes axon regeneration after spinal cord injury","volume":"113","author":"He","year":"2016","journal-title":"Proc. Natl. Acad. Sci."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"653","DOI":"10.1002\/jnr.23733","article-title":"Autophagy inhibition in endogenous and nutrient-deprived conditions reduces dorsal root ganglia neuron survival and neurite growth in vitro","volume":"94","author":"Clarke","year":"2016","journal-title":"J. Neurosci. Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"699","DOI":"10.1038\/s41589-019-0274-x","article-title":"Glycan sulfation patterns define autophagy flux at axon tip via PTPRsigma-cortactin axis","volume":"15","author":"Sakamoto","year":"2019","journal-title":"Nat. Chem. Biol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"5842","DOI":"10.1073\/pnas.0701402104","article-title":"Unc-51-like kinase 1\/2-mediated endocytic processes regulate filopodia extension and branching of sensory axons","volume":"104","author":"Zhou","year":"2007","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Xie, W., Strong, J.A., and Zhang, J.-M. (2017). Active Nerve Regeneration with Failed Target Reinnervation Drives Persistent Neuropathic Pain. eNeuro, 4.","DOI":"10.1523\/ENEURO.0008-17.2017"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"111","DOI":"10.4103\/joacp.JOACP_389_15","article-title":"Tanezumab: Therapy targeting nerve growth factor in pain pathogenesis","volume":"34","author":"Patel","year":"2018","journal-title":"J. Anaesthesiol. Clin. Pharmacol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1097\/FBP.0000000000000422","article-title":"Anti-NGF treatment can reduce chronic neuropathic pain by changing peripheral mediators and brain activity in rats","volume":"30","author":"Evangelista","year":"2019","journal-title":"Behav. Pharmacol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"626","DOI":"10.1038\/s41419-018-0682-y","article-title":"ATG5 overexpression is neuroprotective and attenuates cytoskeletal and vesicle-trafficking alterations in axotomized motoneurons","volume":"9","author":"Bosch","year":"2018","journal-title":"Cell Death Dis."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"983","DOI":"10.1038\/nm.3232","article-title":"The role of autophagy in neurodegenerative disease","volume":"19","author":"Nixon","year":"2013","journal-title":"Nat. Med."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1016\/j.neulet.2015.05.046","article-title":"Increased autophagic activity in dorsal root ganglion attenuates neuropathic pain following peripheral nerve injury","volume":"599","author":"Guo","year":"2015","journal-title":"Neurosci. Lett."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1016\/j.brainres.2013.04.055","article-title":"Expression of LC3 and Beclin 1 in the spinal dorsal horn following spinal nerve ligation-induced neuropathic pain","volume":"1519","author":"Zhang","year":"2013","journal-title":"Brain Res."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1016\/j.pain.2013.09.013","article-title":"Schwann cell autophagy counteracts the onset and chronification of neuropathic pain","volume":"155","author":"Marinelli","year":"2014","journal-title":"Pain"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1038\/s12276-019-0302-7","article-title":"Mechanisms and disease implications of sirtuin-mediated autophagic regulation","volume":"51","author":"Lee","year":"2019","journal-title":"Exp. Mol. Med."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"130","DOI":"10.1097\/j.pain.0000000000000739","article-title":"SIRT1 attenuates neuropathic pain by epigenetic regulation of mGluR1\/5 expressions in type 2 diabetic rats","volume":"158","author":"Zhou","year":"2017","journal-title":"Pain"},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Shao, H., Xue, Q., Zhang, F., Luo, Y., Zhu, H., Zhang, X., Zhang, H., Ding, W., and Yu, B. (2014). Spinal SIRT1 activation attenuates neuropathic pain in mice. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0100938"},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Tang, X., Drotar, J., Li, K., Clairmont, C.D., Brumm, A.S., Sullins, A.J., Wu, H., Liu, X.S., Wang, J., and Gray, N.S. (2019). Pharmacological enhancement of KCC2 gene expression exerts therapeutic effects on human Rett syndrome neurons and Mecp2 mutant mice. Sci. Transl. Med., 11.","DOI":"10.1126\/scitranslmed.aau0164"},{"key":"ref_51","first-page":"5102","article-title":"Effect of Acamprosate in experimental models of peripheral neuropathic and inflammatory pain in wistar rats","volume":"6","author":"Khanna","year":"2015","journal-title":"Int. J. Pharm. Sci. reserach"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"11356","DOI":"10.1523\/JNEUROSCI.6265-11.2012","article-title":"Activity-dependent cleavage of the K-Cl cotransporter KCC2 mediated by calcium-activated protease calpain","volume":"32","author":"Puskarjov","year":"2012","journal-title":"J. Neurosci."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"268","DOI":"10.1002\/jnr.10552","article-title":"Ribavirin reduces clinical signs and pathological changes of experimental autoimmune encephalomyelitis in Dark Agouti rats","volume":"72","author":"Milicevic","year":"2003","journal-title":"J. Neurosci. Res."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1254\/jphs.12004FP","article-title":"The effect of ribavirin on reactive astrogliosis in experimental autoimmune encephalomyelitis","volume":"119","author":"Lavrnja","year":"2012","journal-title":"J. Pharmacol. Sci."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"1661","DOI":"10.1007\/s00705-017-3291-7","article-title":"Ribavirin attenuates the respiratory immune responses to influenza viral infection in mice","volume":"162","author":"Liao","year":"2017","journal-title":"Arch. Virol."},{"key":"ref_56","doi-asserted-by":"crossref","unstructured":"Dai, J.-P., Wang, Q.-W., Su, Y., Gu, L.-M., Zhao, Y., Chen, X.-X., Chen, C., Li, W.-Z., Wang, G.-F., and Li, K.-S. (2017). Emodin Inhibition of Influenza A Virus Replication and Influenza Viral Pneumonia via the Nrf2, TLR4, p38\/JNK and NF-kappaB Pathways. Molecules, 22.","DOI":"10.3390\/molecules22101754"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"573","DOI":"10.1523\/JNEUROSCI.3295-09.2010","article-title":"P2X7-dependent release of interleukin-1beta and nociception in the spinal cord following lipopolysaccharide","volume":"30","author":"Clark","year":"2010","journal-title":"J. Neurosci."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/9\/4\/808\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T09:12:14Z","timestamp":1760173934000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/9\/4\/808"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,3,27]]},"references-count":57,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2020,4]]}},"alternative-id":["cells9040808"],"URL":"https:\/\/doi.org\/10.3390\/cells9040808","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,3,27]]}}}