{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,21]],"date-time":"2026-08-21T05:23:40Z","timestamp":1787289820618,"version":"build-2736575974"},"reference-count":29,"publisher":"MDPI AG","issue":"21","license":[{"start":{"date-parts":[[2025,10,27]],"date-time":"2025-10-27T00:00:00Z","timestamp":1761523200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Lyon Research Aging Program"}],"content-domain":{"domain":["www.mdpi.com"],"crossmark-restriction":true},"short-container-title":["Int. J. Mol. Sci.","IJMS"],"abstract":"<jats:p>Chronic pain is a serious health issue, often irrationally managed by conventional analgesics. Duloxetine, a serotonin\u2013norepinephrine reuptake inhibitor (SNRI), also effective in neuropathic and musculoskeletal pain, but the molecular mechanism of its analgesic action is still unclear. Here, we examined whether Duloxetine exerts pleiotropic effects by directly targeting phosphorylated STAT3 (pSTAT3), a key regulator of neuroinflammation and pain sensitization. Molecular docking showed that Duloxetine binds with pSTAT3 with binding energy \u22125.83 kcal\/mol. Ruxolitinib, a JAK\/STAT inhibitor used as reference, showed binding energy of \u22126.19 kcal\/mol. Molecular dynamics (MD) simulations confirmed stable Duloxetine\u2013pSTAT3 complexes, while MM-PBSA free energy analysis revealed more favorable binding for Duloxetine (\u0394G = \u221215.17 kJ\u00b7mol\u22121) than Ruxolitinib (\u0394G = \u221212.98 kJ\u00b7mol\u22121) for pSTAT3. In-vitro analyses, Western blot showed that Duloxetine significantly reduced IL-6\u2013induced STAT3 and pSTAT3 expression in C2C12 cells in a dose-dependent manner (6.4 and 12.8 \u03bcM, *** p &lt; 0.0001), although Ruxolitinib produced a stronger suppression. Transcriptomic analysis revealed Duloxetine-specific enrichment of mitochondrial, oxidative phosphorylation, and synaptic pathways, distinct from the immune-suppressive influence of Ruxolitinib. RNA-seq further revealed that STAT3 transcript abundance remains constant under all treatment conditions, indicating that post-transcriptional or post-translational mechanisms, such as phosphorylation-dependent activation, may be involved rather than transcriptional modulation of STAT3 in action of Ruxolitinib and Duloxetine and the formation of novel STAT3 indicating enhanced transcript diversity. The rMATS splicing analysis confirmed dose-dependent modulation, with Duloxetine promoting mild exon skipping at 6.4 \u03bcM (IncLevel 0.90 \u2192 0.80) and recovery at 12.8 \u03bcM (0.85 \u2192 0.86), while Ruxolitinib induced stronger exon inclusion (0.85 \u2192 1.00,0.94), with broader transcript suppression at 6.4 \u03bcM and 12.8 \u03bcM, respectively. These findings establish Duloxetine as a dual-action therapeutic that combines neurotransmitter reuptake inhibition with pSTAT3 suppression and isoform-level transcriptomic modulation. This pleiotropic mechanism provides a rationale for its durable analgesic effects and supports repurposing in STAT3-associated disorders.<\/jats:p>","DOI":"10.3390\/ijms262110432","type":"journal-article","created":{"date-parts":[[2025,10,28]],"date-time":"2025-10-28T09:16:07Z","timestamp":1761642967000},"page":"10432","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["RETRACTED: Duloxetine, an SNRI, Targets pSTAT3 Signaling: In-Silico, RNA-Seq and In-Vitro Evidence for a Pleiotropic Mechanism of Pain Relief"],"prefix":"10.3390","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3339-3264","authenticated-orcid":false,"given":"Sayed","family":"Abdi","sequence":"first","affiliation":[{"name":"Department of Geriatrics, Donald W. Reynolds Institute on Aging, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gohar","family":"Azhar","sequence":"additional","affiliation":[{"name":"Department of Geriatrics, Donald W. Reynolds Institute on Aging, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaomin","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Geriatrics, Donald W. Reynolds Institute on Aging, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeanne","family":"Wei","sequence":"additional","affiliation":[{"name":"Department of Geriatrics, Donald W. Reynolds Institute on Aging, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,10,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1261","DOI":"10.1021\/acschemneuro.2c00780","article-title":"Duloxetine and Amitriptyline Reduce Neuropathic Pain by Inhibiting Primary Sensory Input to Spinal Dorsal Horn Neurons via \u03b11- and \u03b12-Adrenergic Receptors","volume":"14","author":"Huang","year":"2023","journal-title":"ACS Chem. Neurosci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1016\/j.lfs.2018.05.025","article-title":"Neuroprotective Effect of Duloxetine in a Mouse Model of Diabetic Neuropathy: Role of Glia Suppressing Mechanisms","volume":"205","author":"Tawfik","year":"2018","journal-title":"Life Sci."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Hayashida, K., and Obata, H. (2019). Strategies to Treat Chronic Pain and Strengthen Impaired Descending Noradrenergic Inhibitory System. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20040822"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1111\/j.1471-4159.2008.05566.x","article-title":"JAK\/STAT3 Pathway Is Activated in Spinal Cord Microglia after Peripheral Nerve Injury and Contributes to Neuropathic Pain Development in Rat","volume":"107","author":"Dominguez","year":"2008","journal-title":"J. Neurochem."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Liu, X., Tian, Y., Lu, N., Gin, T., Cheng, C.H.K., and Chan, M.T.V. (2013). Stat3 Inhibition Attenuates Mechanical Allodynia through Transcriptional Regulation of Chemokine Expression in Spinal Astrocytes. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0075804"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Park, J., Lee, C., and Kim, Y.T. (2023). Effects of Natural Product-Derived Compounds on Inflammatory Pain via Regulation of Microglial Activation. Pharmaceuticals, 16.","DOI":"10.3390\/ph16070941"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Sun, B., Wu, M., Ru, Y., Meng, Y., Zhang, X., Wang, F., Xia, Z., Yang, L., Zhai, Y., and Li, G. (2025). A Novel Compound DBZ Alleviates Chronic Inflammatory Pain and Anxiety-like Behaviors by Targeting the JAK2-STAT3 Signaling Pathway. J. Biol. Chem., 301.","DOI":"10.1016\/j.jbc.2025.110223"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"114576","DOI":"10.1016\/j.expneurol.2023.114576","article-title":"IL-6\/JAK2\/STAT3 Axis Mediates Neuropathic Pain by Regulating Astrocyte and Microglia Activation after Spinal Cord Injury","volume":"370","author":"Lee","year":"2023","journal-title":"Exp. Neurol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1631\/jzus.B1500297","article-title":"STAT3 Signal That Mediates the Neural Plasticity Is Involved in Willed-Movement Training in Focal Ischemic Rats","volume":"17","author":"Tang","year":"2016","journal-title":"J. Zhejiang Univ. Sci. B"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"S10","DOI":"10.1016\/j.pain.2013.06.022","article-title":"Glia and Pain: Is Chronic Pain a Gliopathy?","volume":"154","author":"Ji","year":"2013","journal-title":"Pain"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Abdi, S.A.H., Ali, A., Sayed, S.F., Nagarajan, S., Alam, P., and Ali, A. (2022). Sunscreen Ingredient Octocrylene\u2019s Potency to Disrupt Vitamin D Synthesis. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms231710154"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"104060","DOI":"10.1016\/j.mvr.2020.104060","article-title":"Ruxolitinib Attenuates Intimal Hyperplasia via Inhibiting JAK2\/STAT3 Signaling Pathway Activation Induced by PDGF-BB in Vascular Smooth Muscle Cells","volume":"132","author":"Song","year":"2020","journal-title":"Microvasc. Res."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Meng, J., Zhang, Q., Yang, C., Xiao, L., Xue, Z., and Zhu, J. (2019). Duloxetine, a Balanced Serotonin-Norepinephrine Reuptake Inhibitor, Improves Painful Chemotherapy-Induced Peripheral Neuropathy by Inhibiting Activation of p38 MAPK and NF-\u03baB. Front. Pharmacol., 10.","DOI":"10.3389\/fphar.2019.00365"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"369","DOI":"10.1038\/s41419-024-06749-9","article-title":"A Novel Function of STAT3\u03b2 in Suppressing Interferon Response Improves Outcome in Acute Myeloid Leukemia","volume":"15","author":"Edtmayer","year":"2024","journal-title":"Cell Death Dis."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1864","DOI":"10.1038\/s41386-025-02219-8","article-title":"Chronic Treatment with Fluoxetine Regulates Mitochondrial Features and Plasticity-Associated Transcriptomic Pathways in Parvalbumin-Positive Interneurons of Prefrontal Cortex","volume":"50","author":"Jetsonen","year":"2025","journal-title":"Neuropsychopharmacology"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Rossetti, A.C., Paladini, M.S., Br\u00fcning, C.A., Spero, V., Cattaneo, M.G., Racagni, G., Papp, M., Riva, M.A., and Molteni, R. (2022). Involvement of the IL-6 Signaling Pathway in the Anti-Anhedonic Effect of the Antidepressant Agomelatine in the Chronic Mild Stress Model of Depression. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms232012453"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"769","DOI":"10.1016\/j.pharep.2018.02.016","article-title":"S-Allyl Cysteine Ameliorates Cyclophosphamide-Induced Downregulation of Urothelial Uroplakin IIIa with a Concomitant Effect on Expression and Release of CCL11and TNF-\u03b1 in Mice","volume":"70","author":"Abdi","year":"2018","journal-title":"Pharmacol. Rep."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1087","DOI":"10.62347\/WMQV6643","article-title":"Duloxetine Enhances the Sensitivity of Non-Small Cell Lung Cancer Cells to EGFR Inhibitors by REDD1-Induced MTORC1\/S6K1 Suppression","volume":"14","author":"Jang","year":"2024","journal-title":"Am. J. Cancer Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"354","DOI":"10.1016\/j.intimp.2017.11.032","article-title":"Repression of Interferon \u03b2-Regulated Cytokines by the JAK1\/2 Inhibitor Ruxolitinib in Inflammatory Human Macrophages","volume":"54","author":"Lecureur","year":"2018","journal-title":"Int. Immunopharmacol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1503\/jpn.170199","article-title":"ER\u03b2 Agonist Alters RNA Splicing Factor Expression and Has a Longer Window of Antidepressant Effectiveness than Estradiol after Long-Term Ovariectomy","volume":"44","author":"Hou","year":"2019","journal-title":"J. Psychiatry Neurosci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"180","DOI":"10.4306\/pi.2012.9.2.180","article-title":"Fluoxetine Increases the Expression of NCAM140 and PCREB in Rat C6 Glioma Cells","volume":"9","author":"Choi","year":"2012","journal-title":"Psychiatry Investig."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Mazumder, R., Morampudi, K.S., Motwani, M., Vasudevan, S., and Goldman, R. (2012). Proteome-Wide Analysis of Single-Nucleotide Variations in the N-Glycosylation Sequon of Human Genes. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0036212"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"349","DOI":"10.1080\/15376516.2022.2137872","article-title":"Potential of Paracetamol for Reproductive Disruption: Molecular Interaction, Dynamics, and MM-PBSA Based In-Silico Assessment","volume":"33","author":"Abdi","year":"2023","journal-title":"Toxicol. Mech. Methods"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"598","DOI":"10.1111\/bcpt.12627","article-title":"Cyclophosphamide-induced Down-Regulation of Uroplakin II in the Mouse Urinary Bladder Epithelium Is Prevented by S-Allyl Cysteine","volume":"119","author":"Abdi","year":"2016","journal-title":"Basic Clin. Pharmacol. Toxicol."},{"key":"ref_25","first-page":"36","article-title":"Muscle-Specific Deletion of SOCS3 Increases the Early Inflammatory Response but Does Not Affect Regeneration after Myotoxic Injury. Skelet","volume":"6","author":"Swiderski","year":"2016","journal-title":"Muscle"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Varet, H., Brillet-Gu\u00e9guen, L., Copp\u00e9e, J.-Y., and Dillies, M.-A. (2016). SARTools: A DESeq2- and EdgeR-Based R Pipeline for Comprehensive Differential Analysis of RNA-Seq Data. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0157022"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"304","DOI":"10.12688\/f1000research.23297.1","article-title":"GFF Utilities: GffRead and GffCompare","volume":"9","author":"Pertea","year":"2020","journal-title":"F1000Research"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"E5593","DOI":"10.1073\/pnas.1419161111","article-title":"RMATS: Robust and Flexible Detection of Differential Alternative Splicing from Replicate RNA-Seq Data","volume":"111","author":"Shen","year":"2014","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Abdi, S.A.H., Azhar, G., Zhang, X., Sharma, S., Hafeez, M., and Wei, J.Y. (2025). Alternative Splicing of Serum Response Factor Reveals Isoform-Specific Remodeling in Cardiac Diseases. Genes, 16.","DOI":"10.3390\/genes16080947"}],"updated-by":[{"DOI":"10.3390\/ijms27135656","type":"retraction","label":"Retraction","source":"publisher","updated":{"date-parts":[[2025,10,27]],"date-time":"2025-10-27T00:00:00Z","timestamp":1761523200000}},{"DOI":"10.3390\/ijms27135656","type":"retraction","label":"Retraction","source":"publisher","updated":{"date-parts":[[2026,6,23]],"date-time":"2026-06-23T00:00:00Z","timestamp":1782172800000}}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/21\/10432\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,21]],"date-time":"2026-08-21T04:26:58Z","timestamp":1787286418000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/21\/10432"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10,27]]},"references-count":29,"journal-issue":{"issue":"21","published-online":{"date-parts":[[2025,11]]}},"alternative-id":["ijms262110432"],"URL":"https:\/\/doi.org\/10.3390\/ijms262110432","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,10,27]]}}}