A subscription to JoVE is required to view this content. Sign in or start your free trial.

Research Article

Scalp Acupuncture Combined with Exercise Induces PI3K/Akt Activation and Improves Synaptic Plasticity in Post-Stroke Spastic Rats

115 views

DOI:

10.3791/71433

July 14th, 2026

In This Article

Summary

This study aims to investigate the effects of scalp acupuncture combined with exercise on motor cortex synaptic plasticity in a rat model of post-stroke limb spasticity and to explore the association between PI3K/Akt pathway activity and the therapeutic effects.

Abstract

Effective treatments for post-stroke spasticity (PSS) remain limited, and the mechanism by which scalp acupuncture combined with exercise (SAE) alleviates PSS is not fully understood. This study investigated the effects of SAE in rats with PSS and explored whether SAE regulates synaptic plasticity through the PI3K/Akt pathway. A middle cerebral artery occlusion (MCAO) rat model was established, and rats were randomly assigned to five groups: Blank, Sham, MCAO, SAE, and Baclofen. All interventions were administered for 7 consecutive days. Neurological function, cerebral infarct volume, histopathology, synaptic ultrastructure, and related protein expression were then evaluated. Compared with the MCAO group, SAE improved neurological deficits, reduced muscle hypertonia, decreased infarct volume, alleviated neuronal injury, increased synapse number, and improved synaptic morphology. SAE also upregulated glial cell line-derived neurotrophic factor (GDNF), phosphorylated PI3K (p-PI3K), phosphorylated Akt (p-AKT), and synaptic proteins, including synaptophysin (SYN), postsynaptic density protein 95 (PSD-95), and growth-associated protein 43 (GAP-43). Immunofluorescence staining further suggested enhanced expression of SYN and p-Akt in the motor cortex after SAE treatment. These findings suggest that SAE may be correlated with the activation of PI3K/Akt signaling via upregulation of GDNF, thereby improving synaptic plasticity and alleviating PSS symptoms. This study investigated the effects of SAE in rats with PSS and explored the potential association between SAE intervention, PI3K/Akt pathway activity, and synaptic plasticity.

Introduction

Cerebral infarction remains a leading cause of death and disability worldwide. Ischemia and hypoxia damage neurons, disrupt the excitation–inhibition balance in motor circuits, and lead to motor dysfunction1. Post-stroke spasticity is manifested as increased muscle tone and reduced joint mobility, which impairs activities of daily living and increases caregiver burden2. Although antispastic agents such as baclofen can provide temporary symptomatic relief, long-term use may lead to drug tolerance and adverse effects, and does not restore impaired neural function3. Conventional motor rehabilit....

Access restricted. Please log in or start a trial to view this content.

Protocol

All procedures were approved by the Animal Ethics Committee of Changchun University of Chinese Medicine (Approval No. 20251038) and were conducted in accordance with the Animal Research: Reporting of In Vivo Experiments (ARRIVE 2.0) guidelines as well as the NIH Guide for the Care and Use of Laboratory Animals. All materials used in this study are shown in the Table of Materials.

Experimental procedures
Experimental animals and grouping

Eighty specific pathogen-free (SPF) adult male Sprague–Dawley rats (220–250 g) were purchased from Hongda Animal Breeding Farm (Kuancheng District, C....

Access restricted. Please log in or start a trial to view this content.

Results

SAE improves neurological function and reduces muscle spasticity in PSS rats
The therapeutic effects of SAE on behavioral dysfunction in rats with post-stroke spasticity were evaluated using the Zea Longa score, modified Neurological Severity Score, and modified Ashworth Scale on Days 1, 3, 5, and 7 after model validation (n = 12 per group). The Zea Longa score and mNSS were used to assess neurological deficits, whereas the MAS score was used to evaluate muscle tone/spasticity. After modeling, rats i.......

Access restricted. Please log in or start a trial to view this content.

Discussion

Previous clinical studies have shown that scalp acupuncture combined with exercise can relieve spastic symptoms in patients with post-stroke spasticity, while improving motor function and activities of daily living (ADL)14,24. These findings support its clinical value. The present study further confirmed the therapeutic benefit of SAE in a rat model using three behavioral measures, including the Zea Longa score, mNSS, and MAS. Repeated behavioral assessment showe.......

Access restricted. Please log in or start a trial to view this content.

Disclosures

The authors declare that they have no competing financial interests.

AUTHOR’S CONTRIBUTIONS:
Sainan Zhao: Conceptualization, Methodology, Formal analysis, Investigation, Project administration, Writing – original draft, Writing – review & editing. Lingxu Li: Investigation, Data curation, Validation. Wenna Li: Data curation, Formal analysis, Visualization. Chunmeng Wang: Investigation, Resources. Yanlong Jin: Software, Validation, Writing – review & editing. Guangcheng Ji: Supervision, Funding acquisition, Resources, Writing – review & editing.

Acknowledgements

The authors thank all participants who participated in this study. This work was supported by the Science and Technology Development Project of Jilin Province [No. YDZJ202401122ZYTS].

....

Access restricted. Please log in or start a trial to view this content.

Materials

List of materials used in this article
NameCompanyCatalog NumberComments
2,3,5-triphenyltetrazolium chloride staining solutionBeyotimeC0651TTC staining
3MM Filter PaperWhatman, UK3030861Western blot transfer
AKT AntibodyAffinityAF0836Primary antibody
AcetoneSinopharm Chemical Reagent Co., Ltd.10000418Infiltration of electron microscopy specimens
Adobe IllustratorAdobe Inc.Version 29.5 (64-bit), https://www.adobe.com/products/illustrator.html64-bit version, used for graphic editing and scientific figure preparation
Anhydrous ethanolSinopharm Chemical Reagent Co., Ltd.100092683Deparaffinization of paraffin sections
Anti-fade mounting mediumWuhan Baiqiandu Biotechnology Co., Ltd.C1020coverslipping
BCA Protein Assay KitMDLMD913053Protein concentration determination
Bluishing solutionWuhan Baiqiandu Biotechnology Co., Ltd.C2104Hematoxylin staining
Chemiluminescence Imaging SystemCLINX, ChinaChemiScope6100System Detect target protein
Concentrated normal goat serumBoster Biological TechnologyAR1009serum blocking
DAPIWuhan Baiqiandu Biotechnology Co., Ltd.C1030nuclear staining
Diamond KnifeDaitomeUltra 45°Slice ultrathin specimens
Differentiation solutionWuhan Baiqiandu Biotechnology Co., Ltd.C2104HE staining
Drying OvenTianjin Labory Instruments Equipment Co., Ltd.GFL-230Bake fixed tissue slices
EDTA (pH 9.0) antigen retrieval solutionWuhan Baiqiandu Biotechnology Co., Ltd.C1052Antigen retrieval
Electron microscopy fixativeASPENAS1063Fixation of electron microscopy specimens
Electrophoresis Power SupplyLongfangLF-2000Provide electrophoresis voltage
Embed 812 embedding mediumSPI90529-77-4Infiltration of electron microscopy specimens
EndNote X9Clarivate Analytics (US) LLCVersion X9, https://endnote.com/Used for reference management and academic citation formatting
Freezing StageWuhan Junjie Electronics Co., Ltd.JB-L5Freeze frozen tissue blocks
GAP-43 AntibodyAffinityDf7766Primary antibody
GAPDH AntibodyAffinityAF7021Primary antibody
GDNF AntibodyBiosynthesisBs-1024rPrimary antibody
Glacial acetic acidWuhan Baiqiandu Biotechnology Co., Ltd.C2138Nissl staining
Glass Slides and CoverslipsSinopharm Chemical Reagent Co., Ltd.10212432CMount tissue sections
GraphPad PrismGraphPad Software, LLCVersion 10.1.2 (324) for Windows 64-bit, https://www.graphpad.com/64-bit version, used for statistical analysis and scientific plotting
Grinding machineBeijing HerdeN.9548Grind tissue samples
HE staining solutionWuhan Baiqiandu Biotechnology Co., Ltd.C2103HE staining
HRP-Conjugated Goat Anti-Mouse Secondary AntibodyServicebioC030205Secondary Antibody
HRP-conjugated goat anti-rabbit antibodySeraCare5220-0336Secondary antibody
Hydrogen peroxide (H2O2)Wuhan Baiqiandu Biotechnology Co., Ltd.C5101preparation of TSA reagents
IBM SPSS StatisticsIBM Corp.Version 25, 64-bit, https://www.ibm.com/products/spss-statistics64-bit version, used for statistical data analysis and processing
Imaging SystemNikon, JapanNikon DS-U3Capture microscopic images
Metal BathCOYOTEH203-HConstant temperature incubation
Microwave OvenGalanz Microwave Electrical Appliance Co., Ltd.P70D20TL-P4Antigen retrieval
NC Membrane (0.22 μm)GVS, USAISEQ00010Protein blot transfer
Neutral balsamSinopharm Chemical Reagent Co., Ltd.10004160dehydration and coverslipping
PBS bufferWuhan Baiqiandu Biotechnology Co., Ltd.C0003decolorization and washing
PI3K AntibodyAffinityAF6241Primary antibody
PSD-95 AntibodyAffinityAf5283Primary antibody
Pathological MicrotomeLeica Instruments (Shanghai) Co., Ltd.RM2016Cut paraffin sections
Phospho-AKT AntibodyAffinityAf0832Primary antibody
Phospho-PI3K AntibodyAffinityAf3241Primary antibody
PipetteDragonlabKE0003087/KA0056573Quantitative liquid transfer
Protease Inhibitor CocktailBeyotimeP1005Protease inhibition
RIPA Lysis Buffer (Medium)BeyotimeP0013CTissue lysis
Refrigerated CentrifugeSEILOGEX, ChinaCF1524RLow-speed sample separation
SDS-PAGE Electrophoresis SystemBeyotimeMiniProGel™ Gel Casting, Electrophoresis and Western Blot Transfer SystemProtein electrophoretic separation
SDS-PAGE Precast Gel KitMDLMD911919SDS-PAGE electrophoresis
SYN (Synapsin) AntibodyBiosynthesisbs-3501RPrimary antibody
ShakerQilinbeierTS-8Sample oscillation washing
Tissue DehydratorWuhan Junjie Electronics Co., Ltd.JJ-12JDehydrate paraffin tissue
Tissue Embedding MachineWuhan Junjie Electronics Co., Ltd.JB-P5Embed tissue in paraffin
Tissue Spreading MachineWuhan Junjie Electronics Co., Ltd.KD-PUnfold paraffin slices
Toluidine blue staining solutionWuhan Baiqiandu Biotechnology Co., Ltd.C2131Nissl staining
Transmission Electron MicroscopeFEITecnai G2 20 TWINObserve ultrastructure
Tri-Color Pre-Stained Broad Range Protein Molecular Weight Marker (10–180 kDa)ServicebioG2091SDS-PAGE electrophoresis
Tyramide-488Wuhan Baiqiandu Biotechnology Co., Ltd.C6002TSA reagents
Tyramide-CY3Wuhan Baiqiandu Biotechnology Co., Ltd.C6003TSA reagents
UltramicrotomeLeicaLeica UC7Cut ultrathin EM sections
Ultrasonic Cell DisruptorXianchangXC-CDLyse cells
Upright Optical MicroscopeNikon, JapanNikon Eclipse ciObserve stained slices
Vortex MixerTianyue ElectronicsTYXH-IIMix liquid samples
Western Blot Transfer TankJunyi DongfangJY-ZY6Protein membrane transfer
XyleneSinopharm Chemical Reagent Co., Ltd.10023418Deparaffinization of paraffin sections

Reprints and Permissions

Tags

NeuroscienceMotion style scalp acupuncturecerebral infarctionlimb spasticityPI3K Akt pathwayGDNF