10.7910/DVN/FGUGCH Liping Wu https://orcid.org/0009-0007-8844-654X The First Clinical Medical College, Guangxi University of Chinese Medicine, Nanning, Guangxi Zhuang Autonomous Region, China Xiang Wang Xiang Wang https://orcid.org/0009-0006-3460-666X Department of Medicine, Saines College of New Medicine, Guangxi University of Chinese Medicine, Nanning, Guangxi Zhuang Autonomous Region, China Peng Yang Peng Yang https://orcid.org/0000-0003-2904-4202 Department of Tuina, Fangchenggang Hospital, Guangxi University of Chinese Medicine, Fangchenggang, Guangxi Zhuang Autonomous Region, China Xiongjiang Wang https://orcid.org/0009-0004-1476-4770 The First Clinical Medical College, Guangxi University of Chinese Medicine, Nanning, Guangxi Zhuang Autonomous Region, China Yujiao Zheng The First Clinical Medical College, Guangxi University of Chinese Medicine, Nanning, Guangxi Zhuang Autonomous Region, China Yeni Tang https://orcid.org/0009-0007-3295-3657 The First Clinical Medical College, Guangxi University of Chinese Medicine, Nanning, Guangxi Zhuang Autonomous Region, China Haoxin Zheng https://orcid.org/0009-0002-8613-5864 The First Clinical Medical College, Guangxi University of Chinese Medicine, Nanning, Guangxi Zhuang Autonomous Region, China Peng Ning Peng Ning https://orcid.org/0009-0002-8613-5864 The First Clinical Medical College, Guangxi University of Chinese Medicine, Nanning, Guangxi Zhuang Autonomous Region, China Peng Ning Peng Ning https://orcid.org/0009-0005-1863-6431 The First Clinical Medical College, Guangxi University of Chinese Medicine, Nanning, Guangxi Zhuang Autonomous Region, China Hongliang Tang https://orcid.org/0000-0002-3021-5585 Department of Tuina, Fangchenggang Hospital, Guangxi University of Chinese Medicine, Fangchenggang, Guangxi Zhuang Autonomous Region, China Harvard Dataverse 2026 Medicine, Health and Life Sciences Epigenesis, Genetic Musculoskeletal Manipulations Neuralgia RNA Methylation Suppressor of Cytokine Signaling 1 Protein Toll-Like Receptor 4 Hongliang Tang Department of Tuina, Fangchenggang Hospital, Guangxi University of Chinese Medicine, Fangchenggang, Guangxi Zhuang Autonomous Region, China Liping Wu The First Clinical Medical College, Guangxi University of Chinese Medicine, Nanning, Guangxi Zhuang Autonomous Region, China 2026-02-08 2026-03-13 182353 207555 206780 44769 165592 55249 application/octet-stream application/octet-stream application/octet-stream application/octet-stream application/octet-stream application/octet-stream 1.0 Creative Commons CC0 1.0 Universal Public Domain Dedication. Background: Manipulation therapy (MT) relieves pain in nervous system disorders by reducing inflammation. Although evidence suggests MT is beneficial for neuropathic pain (NP), its molecular mechanisms remain unclear. This study investigated whether MT could reverse NP-related epigenetic changes and elucidated the potential mechanisms of NP. Methods: An NP model was established via spinal nerve ligation (SNL), and MT was administered at acupoints Jumping Round (GB30), Yang Mound Spring (GB34), and Suspended Bell (GB39) for 14 days. Mechanical and thermal pain in rats were evaluated using the mechanical paw withdrawal threshold and thermal paw withdrawal latency tests. Inflammatory cytokine levels in spinal cord neurons were quantified using enzyme-linked immunosorbent assay. Total N6-methyladenosine (m6A) levels were assessed by colorimetric analysis, while target genes were validated using methylated RNA immunoprecipitation succeeded by quantitative polymerase chain reaction, immunofluorescence, Western blotting, and quantitative real-time polymerase chain reaction. Results: The outcomes indicated that MT significantly attenuated SNL-induced pain hypersensitivity. The therapeutic effects of MT were correlated with global m6A methylation in spinal cord neurons. Moreover, MT affected the toll-like receptor 4 (TLR4) signaling pathway’s expression by modulating suppressor of cytokine signaling 1 protein (SOCS1) mRNA m6A methylation levels, consequently reducing the secretion of neuroinflammatory cytokines interleukin-1 beta (IL-1β), IL-6, and tumor necrosis factor-alpha (TNF-α), thereby alleviating NP.