Spinal Tuina ameliorates cerebral palsy-associated neural impairment in hypoxia-ischemia model rats via BMSC-derived exosomal microRNAs.

BACKGROUND

Spinal Tuina is effective for cerebral palsy (CP), but its molecular mechanism remains unclear. Prior studies indicate that acupoint stimulation regulates miRNAs in bone marrow mesenchymal stem cell-derived exosomes (BMSCs-Exo) to promote brain repair. This study explores whether Spinal Tuina treats CP via a similar mechanism.

METHODS

A neonatal hypoxic-ischemic (HI) model was established in 3-day-old Sprague-Dawley rats via left common carotid artery ligation and hypoxia, followed by standardized Tuina. BMSCs were extracted via adherence separation, and exosomes were isolated by ultracentrifugation. Tuina's effects on BMSC proliferation (colony formation assay) and exosome release nanoparticle tracking analysis, were assessed. High-throughput miRNA sequencing, qRT-PCR, and bioinformatics identified differentially expressed miRNAs and their regulatory pathways.

RESULTS

HI + Tuina rats showed improved affected-limb motor function. BMSCs from the HI + Tuina group exhibited elevated fibroblast colony-forming units and exosome concentrations compared to HI Untreated and Sham groups. Among differentially expressed miRNAs in BMSCs-Exo, 14 were upregulated in HI + Tuina vs. HI Untreated, with mir-150-5p highlighted as a key regulator. Kyoto encyclopedia of genes and genomes analysis revealed target gene enrichment in mitogen-activated protein kinase (MAPK), tumor necrosis factor (TNF), axon guidance, and Hippo signaling pathways.

CONCLUSION

Spinal Tuina improves motor function in HI rats, enhances BMSC proliferation and exosome release, and regulates exosomal miRNAs. MiR-150-5p is a critical mediator, with differentially expressed miRNAs potentially modulating inflammation, neurodevelopment, and cell survival in HI-induced CP.