Constraint-induced movement therapy combined with mesenchymal stem cell transplantation promotes myelination and functional recovery by inhibiting PRKCD/MEK/ERK pathway in hemiplegic cerebral palsy rats

Abstract Background The core problem of hemiplegic cerebral palsy (HCP) is upper limb motor deficits with high rates of disability. Given the shared goals of stem cell therapy and rehabilitation, this study investigated the synergistic effects of constraint-induced movement therapy (CIMT) and human...

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Main Authors: Xiaolin Guo, Liru Liu, Jie Luo, Tingting Peng Jr., You Wang, Shiya Huang, Xiaoli Zeng, Tingting Peng Sr., Aihua Chen, Mengru Zhong, Yage Zhang, Kaishou Xu, Lu He
Format: Article
Language:English
Published: BMC 2025-08-01
Series:Stem Cell Research & Therapy
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Online Access:https://doi.org/10.1186/s13287-025-04544-7
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author Xiaolin Guo
Liru Liu
Jie Luo
Tingting Peng Jr.
You Wang
Shiya Huang
Xiaoli Zeng
Tingting Peng Sr.
Aihua Chen
Mengru Zhong
Yage Zhang
Kaishou Xu
Lu He
author_facet Xiaolin Guo
Liru Liu
Jie Luo
Tingting Peng Jr.
You Wang
Shiya Huang
Xiaoli Zeng
Tingting Peng Sr.
Aihua Chen
Mengru Zhong
Yage Zhang
Kaishou Xu
Lu He
author_sort Xiaolin Guo
collection DOAJ
description Abstract Background The core problem of hemiplegic cerebral palsy (HCP) is upper limb motor deficits with high rates of disability. Given the shared goals of stem cell therapy and rehabilitation, this study investigated the synergistic effects of constraint-induced movement therapy (CIMT) and human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) transplantation in promoting motor recovery and elucidates the underlying mechanisms in HCP. Methods The rats were allocated to a control group and HCP groups receiving different interventions (CIMT, hUC-MSCs or combination treatment), namely, the Control, HCP, HCP + CIMT, HCP + MSC, and HCP + CIMT + MSC groups. Motor function was evaluated using rotarod duration, grip strength, and forelimb suspension time. Golgi-Cox staining, transmission electron microscopy, immunofluorescence staining, Western blotting, quantitative real-time PCR and label-free proteomic quantification technology were used to measure dendritic/axonal area, myelin integrity, oligodendrocyte (OL)/oligodendrocyte precursor cell (OPC)-associated proteins and PRKCD/MEK/ERK expression in the motor cortex. Results Rats in the HCP + CIMT + MSC group exhibited improved motor function, increased dendritic spines and branches, enhanced myelin integrity, higher numbers of Olig2+OL, CNPase+OL and MBP+OL, and reduced NG2+OPC counts in the motor cortex compared to the HCP group (p < 0.05). Additionally, motor performance in the HCP + CIMT + MSC group was significantly superior to than those in the HCP + CIMT and HCP + MSC groups (p < 0.05). Moreover, proteomic analysis identified that PRKCD, a key mediator of the synergistic effect, was expressed in OPC and implicated in their physiological processes. Rats in the HCP + CIMT + MSC group also showed reduced PRKCD protein and mRNA expression, fewer PRKCD+/NG2+ cells, higher CNPase+/PRKCD+ area ratio and lower levels of MEK1/2 and ERK1/2 phosphorylation relative to the HCP group (p < 0.05). Conclusions CIMT combined with hUC-MSCs transplantation synergistically promoted OPC differentiation into immature OL, induced myelination, and restored motor function in HCP rats, potentially by inhibiting the PRKCD/MEK/ERK pathway. This combined approach expands therapeutic options for HCP and identifies a promising target for future interventions.
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spelling doaj-art-d95285b043d0485481df09c69868e7282025-08-24T11:11:09ZengBMCStem Cell Research & Therapy1757-65122025-08-0116112010.1186/s13287-025-04544-7Constraint-induced movement therapy combined with mesenchymal stem cell transplantation promotes myelination and functional recovery by inhibiting PRKCD/MEK/ERK pathway in hemiplegic cerebral palsy ratsXiaolin Guo0Liru Liu1Jie Luo2Tingting Peng Jr.3You Wang4Shiya Huang5Xiaoli Zeng6Tingting Peng Sr.7Aihua Chen8Mengru Zhong9Yage Zhang10Kaishou Xu11Lu He12Department of Rehabilitation, Guangzhou Women and Children’s Medical Center, Guangzhou Medical UniversityDepartment of Rehabilitation, Guangzhou Women and Children’s Medical Center, Guangzhou Medical UniversityDepartment of Rehabilitation, Guangzhou Women and Children’s Medical Center, Guangzhou Medical UniversityDepartment of Rehabilitation, Guangzhou Women and Children’s Medical Center, Guangzhou Medical UniversityDepartment of Rehabilitation, Guangzhou Women and Children’s Medical Center, Guangzhou Medical UniversityDepartment of Rehabilitation, Guangzhou Women and Children’s Medical Center, Guangzhou Medical UniversityGuangdong Xiangxue Stem Cell Regenerative Medicine Technology Co., LtdDepartment of Rehabilitation, Guangzhou Women and Children’s Medical Center, Guangzhou Medical UniversityGuangdong Xiangxue Stem Cell Regenerative Medicine Technology Co., LtdDepartment of Rehabilitation, Guangzhou Women and Children’s Medical Center, Guangzhou Medical UniversityDepartment of Rehabilitation, Guangzhou Women and Children’s Medical Center, Guangzhou Medical UniversityDepartment of Rehabilitation, Guangzhou Women and Children’s Medical Center, Guangzhou Medical UniversityDepartment of Rehabilitation, Guangzhou Women and Children’s Medical Center, Guangzhou Medical UniversityAbstract Background The core problem of hemiplegic cerebral palsy (HCP) is upper limb motor deficits with high rates of disability. Given the shared goals of stem cell therapy and rehabilitation, this study investigated the synergistic effects of constraint-induced movement therapy (CIMT) and human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) transplantation in promoting motor recovery and elucidates the underlying mechanisms in HCP. Methods The rats were allocated to a control group and HCP groups receiving different interventions (CIMT, hUC-MSCs or combination treatment), namely, the Control, HCP, HCP + CIMT, HCP + MSC, and HCP + CIMT + MSC groups. Motor function was evaluated using rotarod duration, grip strength, and forelimb suspension time. Golgi-Cox staining, transmission electron microscopy, immunofluorescence staining, Western blotting, quantitative real-time PCR and label-free proteomic quantification technology were used to measure dendritic/axonal area, myelin integrity, oligodendrocyte (OL)/oligodendrocyte precursor cell (OPC)-associated proteins and PRKCD/MEK/ERK expression in the motor cortex. Results Rats in the HCP + CIMT + MSC group exhibited improved motor function, increased dendritic spines and branches, enhanced myelin integrity, higher numbers of Olig2+OL, CNPase+OL and MBP+OL, and reduced NG2+OPC counts in the motor cortex compared to the HCP group (p < 0.05). Additionally, motor performance in the HCP + CIMT + MSC group was significantly superior to than those in the HCP + CIMT and HCP + MSC groups (p < 0.05). Moreover, proteomic analysis identified that PRKCD, a key mediator of the synergistic effect, was expressed in OPC and implicated in their physiological processes. Rats in the HCP + CIMT + MSC group also showed reduced PRKCD protein and mRNA expression, fewer PRKCD+/NG2+ cells, higher CNPase+/PRKCD+ area ratio and lower levels of MEK1/2 and ERK1/2 phosphorylation relative to the HCP group (p < 0.05). Conclusions CIMT combined with hUC-MSCs transplantation synergistically promoted OPC differentiation into immature OL, induced myelination, and restored motor function in HCP rats, potentially by inhibiting the PRKCD/MEK/ERK pathway. This combined approach expands therapeutic options for HCP and identifies a promising target for future interventions.https://doi.org/10.1186/s13287-025-04544-7Combination therapyConstraint-induced movement therapyMesenchymal stem cellsMotor functionOligodendrocyte progenitor cells differentiationMyelination
spellingShingle Xiaolin Guo
Liru Liu
Jie Luo
Tingting Peng Jr.
You Wang
Shiya Huang
Xiaoli Zeng
Tingting Peng Sr.
Aihua Chen
Mengru Zhong
Yage Zhang
Kaishou Xu
Lu He
Constraint-induced movement therapy combined with mesenchymal stem cell transplantation promotes myelination and functional recovery by inhibiting PRKCD/MEK/ERK pathway in hemiplegic cerebral palsy rats
Stem Cell Research & Therapy
Combination therapy
Constraint-induced movement therapy
Mesenchymal stem cells
Motor function
Oligodendrocyte progenitor cells differentiation
Myelination
title Constraint-induced movement therapy combined with mesenchymal stem cell transplantation promotes myelination and functional recovery by inhibiting PRKCD/MEK/ERK pathway in hemiplegic cerebral palsy rats
title_full Constraint-induced movement therapy combined with mesenchymal stem cell transplantation promotes myelination and functional recovery by inhibiting PRKCD/MEK/ERK pathway in hemiplegic cerebral palsy rats
title_fullStr Constraint-induced movement therapy combined with mesenchymal stem cell transplantation promotes myelination and functional recovery by inhibiting PRKCD/MEK/ERK pathway in hemiplegic cerebral palsy rats
title_full_unstemmed Constraint-induced movement therapy combined with mesenchymal stem cell transplantation promotes myelination and functional recovery by inhibiting PRKCD/MEK/ERK pathway in hemiplegic cerebral palsy rats
title_short Constraint-induced movement therapy combined with mesenchymal stem cell transplantation promotes myelination and functional recovery by inhibiting PRKCD/MEK/ERK pathway in hemiplegic cerebral palsy rats
title_sort constraint induced movement therapy combined with mesenchymal stem cell transplantation promotes myelination and functional recovery by inhibiting prkcd mek erk pathway in hemiplegic cerebral palsy rats
topic Combination therapy
Constraint-induced movement therapy
Mesenchymal stem cells
Motor function
Oligodendrocyte progenitor cells differentiation
Myelination
url https://doi.org/10.1186/s13287-025-04544-7
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