Articles
Vol. 70 No. 2 (2026)
https://doi.org/10.4081/ejh.2026.4533

Electroacupuncture combined with rTMS promotes neuronal regeneration via DNMT1-mediated PI3K-AKT pathway in cerebral palsy model

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Published: 20 April 2026
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Electroacupuncture, repetitive transcranial magnetic stimulation, cerebral palsy, neuronal regeneration, DNMT1, epigenetic regulation, PI3K-AKT pathway

Authors

Juan Fan
Tsinghua University Yuquan Hospital, Beijing, China.
Yu Ma
Tsinghua University Yuquan Hospital, Beijing, China.
Liying Sun
Tsinghua University Yuquan Hospital, Beijing, China.
Suhua Miao
Tsinghua University Yuquan Hospital, Beijing, China.
Jianghong He
13581984292@163.com
Tiantan Hospital, Capital Medical University, Beijing, China.

Cerebral palsy (CP), resulting from perinatal brain injury, is characterized by persistent motor dysfunction largely due to impaired regeneration of the corticospinal tract. Electroacupuncture (EA) and repetitive transcranial magnetic stimulation (rTMS) are promising non-invasive neuromodulation approaches, but their combined effects and epigenetic mechanisms remain unclear. This study examined the synergistic neurorestorative potential of EA and rTMS in a hypoxic–ischemic brain damage (HIBD) model of CP, focusing on DNA methyltransferase 1 (DNMT1) regulation of the PI3K-AKT pathway. In vivo, neonatal rats subjected to HIBD were assigned to control, HIBD, EA, rTMS, or combined EA+rTMS groups and treated for four weeks. Motor function was assessed using rotarod, grid-walking, and grip strength tests, while neuronal and molecular changes were analyzed via histological and biochemical methods. In vitro, hypoxia-exposed cortical neurons underwent DNMT1 and PI3K modulation to clarify mechanisms. Combined EA+rTMS produced superior functional recovery compared to monotherapies, including a 2.1-fold increase in rotarod latency, 53% reduction in foot faults, and 80% improvement in grip strength vs untreated HIBD animals. Improvements were also greater than EA or rTMS alone. Histological findings showed enhanced neuronal density in cortical and spinal regions. At the molecular level, combined treatment suppressed DNMT1 expression and increased PI3K, phosphorylated AKT, GAP-43, and myelin basic protein. In vitro, DNMT1 knockdown enhanced PI3K-AKT signaling, neuronal survival, and axonal growth, while DNMT1 overexpression or PI3K inhibition negated these effects. Overall, EA+rTMS promotes functional and structural recovery in CP by epigenetically inhibiting DNMT1 and activating the PI3K-AKT pathway, supporting its translational potential for perinatal brain injury.

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Ethics Approval

All procedures were approved by the Animal Ethics Committee of Tiantan Hospital Animal Center.

CRediT authorship contribution

Juan Fan, Jianghong He,  study design, experiments; Yu Ma, Liying Sun, data collection; Yu Ma, Liying Sun, Suhua Miao, data analysis. Juan Fan, Jianghong He, writing - original draft. All authors read and approved the final manuscript and agreed to be accountable for all aspects of the work

Supporting Agencies

Beijing Natural Science Foundation

Data Availability Statement

The data that support the findings of this study are available from the corresponding author upon reasonable request.

How to Cite



1.
Fan J, Ma Y, Sun L, Miao S, He J. Electroacupuncture combined with rTMS promotes neuronal regeneration via DNMT1-mediated PI3K-AKT pathway in cerebral palsy model. Eur J Histochem [Internet]. 2026 Apr. 20 [cited 2026 Apr. 20];70(2). Available from: https://www.ejh.it/ejh/article/view/4533
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