Application of weighted gene co-expression network analysis to explore the targets of treadmill training in rats with spinal cord injury
- 1Department of Orthopaedics, the General Hospital of Northern Theater Command
Abstract
Background: Spinal cord injury (SCI) describes any damage to the spinal cord that results in a loss of function. With these injuries being commonly treated with subsequent treadmill training (TM), this study aimed to reveal the mechanisms facilitating TM-mediated rehabilitation following a SCI.
Methods: Differentially expressed genes (DEGs) were identified and then subjected to a weighted gene co-expression network analysis to identify TM-associated modules. Co-expression network construction, functional annotation, protein–protein interaction (PPI) network, and transcription factor (TF)-miRNA-target regulatory network were then conducted. We established an SCI rat model and validated key RNAs using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Predicted interactions between miR-135a-5p and JAK2 and STAT1 were tested using dual-luciferase reporter assay.
Results: Total 640 and 877 genes separately differentially expressed in SCI 8d + TM vs. SCI 8d and SCI 14d + TM vs. SCI 14d comparison groups were screened and merged as the DEGs, which were further narrowed to four TM-associated modules linked to the glutamatergic synapse and ribosome biogenesis pathways. PPI network evaluations identified v-Akt murine thymoma viral oncogene homolog 1 (AKT1), Janus kinase 2 (JAK2), tumor necrosis factor (TNF), and signal transducer and activator of transcription 1 (STAT1) as hub nodes. JAK2 and STAT1 were also shown to have interactions in module b. TF-miRNA-target regulatory network revealed that both STAT1 and JAK2 were targets of miR-135a. RT-qPCR results confirmed upregulation of AKT1 and STAT1 and downregulation of JAK2 , TNF and miR-135a after TM. Dual-luciferase reporter assays revealed the direct binding of miR-135a-5p to JAK2 and STAT1 .
Conclusion: These findings suggest that TM facilitates SCI recovery by regulating AKT1 , JAK2 , TNF , STAT1 and miR-135a.
Related
- Optimizing motor functions in incomplete spinal cord injury with simultaneous application of transcranial direct current stimulation and virtual reality assisted treadmill training: a study protocol for a randomized controlled trial {1a}
- Identification of Mitochondrial Dynamics-Related Genes as Diagnostic Biomarkers for Osteoarthritis Using Weighted Gene Co-expression Network Analysis and Machine Learning
- Rhythmic auditory stimulation combined with split-belt treadmill training improves spatiotemporal gait parameters in hemiparetic stroke patients: a randomized controlled trial
- Treadmill prehabilitation attenuates jejunal inflammation and enhances cytoprotective protein expression in murine gut ischemia–reperfusion
- Non-Invasive Assessment of SMAD4 Expression Status in Pancreatic Ductal Adenocarcinoma: Diagnostic Value of T1ρ-Weighted Imaging Combined with Clinical Parameters
BibTeX
@article{zhang2026application,
title = {{Application of weighted gene co-expression network analysis to explore the targets of treadmill training in rats with spinal cord injury}},
author = {Haocong Zhang and Mingming Guo and Lingzhi Meng and Meihui Piao and Hailong Yu},
journal = {Frontiers in Neurology},
year = {2026},
month = oct,
doi = {10.3389/fneur.2026.1846393},
url = {https://www.frontiersin.org/articles/10.3389/fneur.2026.1846393/full},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Application of weighted gene co-expression network analysis to explore the targets of treadmill training in rats with spinal cord injury
AU - Haocong Zhang
AU - Mingming Guo
AU - Lingzhi Meng
AU - Meihui Piao
AU - Hailong Yu
PY - 2026
DA - 2026/10/26
JO - Frontiers in Neurology
DO - 10.3389/fneur.2026.1846393
UR - https://www.frontiersin.org/articles/10.3389/fneur.2026.1846393/full
AB - Background: Spinal cord injury (SCI) describes any damage to the spinal cord that results in a loss of function. With these injuries being commonly treated with subsequent treadmill training (TM), this study aimed to reveal the mechanisms facilitating TM-mediated rehabilitation following a SCI. Methods: Differentially expressed genes (DEGs) were identified and then subjected to a weighted gene co-expression network analysis to identify TM-associated modules. Co-expression network construction, functional annotation, protein–protein interaction (PPI) network, and transcription factor (TF)-miRNA-target regulatory network were then conducted. We established an SCI rat model and validated key RNAs using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Predicted interactions between miR-135a-5p and JAK2 and STAT1 were tested using dual-luciferase reporter assay. Results: Total 640 and 877 genes separately differentially expressed in SCI 8d + TM vs. SCI 8d and SCI 14d + TM vs. SCI 14d comparison groups were screened and merged as the DEGs, which were further narrowed to four TM-associated modules linked to the glutamatergic synapse and ribosome biogenesis pathways. PPI network evaluations identified v-Akt murine thymoma viral oncogene homolog 1 (AKT1), Janus kinase 2 (JAK2), tumor necrosis factor (TNF), and signal transducer and activator of transcription 1 (STAT1) as hub nodes. JAK2 and STAT1 were also shown to have interactions in module b. TF-miRNA-target regulatory network revealed that both STAT1 and JAK2 were targets of miR-135a. RT-qPCR results confirmed upregulation of AKT1 and STAT1 and downregulation of JAK2 , TNF and miR-135a after TM. Dual-luciferase reporter assays revealed the direct binding of miR-135a-5p to JAK2 and STAT1 . Conclusion: These findings suggest that TM facilitates SCI recovery by regulating AKT1 , JAK2 , TNF , STAT1 and miR-135a.
SN - 1664-2295
ER -