Analysis of gene expression profiles and experimental validations of a rat chronic cervical cord compression model

• Published in: Neurochemistry international Vol. 168; p. 105564
• Main Authors: Pu, Pei-min, Li, Zhuo-yao, Dai, Yu-xiang, Sun, Yue-li, Wang, Yong-jun, Cui, Xue-jun, Yao, Min
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.09.2023
• Subjects: Animals; Apoptosis; Astrogliosis; Cervical Cord - pathology; Cervical spondylotic myelopathy; Chronic spinal cord compression; Gliosis - metabolism; Interleukin-17 - metabolism; Interleukin-17 - therapeutic use; Neuroinflammation; Neuroinflammatory Diseases; Phosphatidylinositol 3-Kinases - metabolism; Proto-Oncogene Proteins c-akt - metabolism; Rats; Spinal Cord - metabolism; Spinal Cord Compression - pathology; Spinal Cord Diseases - complications; Spinal Cord Injuries - metabolism; Transcriptome; Transforming Growth Factor beta - metabolism; Astrogliosis; Neuroinflammation; Cervical spondylotic myelopathy; Chronic spinal cord compression; Apoptosis
• ISSN: 0197-0186; 1872-9754; 1872-9754
• DOI: 10.1016/j.neuint.2023.105564

Cervical spondylotic myelopathy (CSM) is a severe non-traumatic spinal cord injury (SCI) wherein the spinal canal and cervical cord are compressed due to the degeneration of cervical tissues. To explore the mechanism of CSM, the ideal model of chronic cervical cord compression in rats was constructed by embedding a polyvinyl alcohol-polyacrylamide hydrogel in lamina space. Then, the RNA sequencing technology was used to screen the differentially expressed genes (DEGs) and enriched pathways among intact and compressed spinal cords. A total of 444 DEGs were filtered out based on the value of log2(Compression/Sham); these were associated with IL-17, PI3K-AKT, TGF-β, and Hippo signaling pathways according to the GSEA, KEGG, and GO analyses. Transmission electron microscopy indicated the changes in mitochondrial morphology. Western blot and immunofluorescence staining revealed neuronal apoptosis, astrogliosis and microglial neuroinflammation in the lesion area. Specifically, the expression of apoptotic indicators, such as Bax and cleaved caspase-3, and inflammatory cytokines, such as IL-1β, IL-6, and TNF-α, were upregulated. The activation of IL-17 signaling pathway was observed in microglia instead of neurons or astrocytes, the activation of TGF-β and inhibition of Hippo signaling pathways were detected in astrocytes instead of neurons or microglia, and the inhibition of PI3K-AKT signaling pathway was discovered in neurons rather than microglia of astrocytes in the lesion area. In conclusion, this study indicated that neuronal apoptosis was accompanied by inhibiting of the PI3K-AKT pathway. Then, the activation of microglia IL-17 pathway and NLRP3 inflammasome effectuated the neuroinflammation, and astrogliosis was ascribed to the activation of TGF-β and the inhibition of the Hippo pathway in the chronic cervical cord of compression. Therefore, therapeutic methods targeting these pathways in nerve cells could be promising CSM treatments. Neuronal apoptosis, neuroinflammation, and astrogliosis were observed and associated with PI3K-AKT, IL-17, TGF-β and Hippo pathways in the cervical cord of rats subject to chronic compression. [Display omitted] •444 DEGs were screened out after cervical cord compression, and were related to inflammatory response and wound healing.•KEGG analysis further identified the DEGs associated with IL-17, PI3K-AKT, TGF-β, and Hippo signaling pathways.•IL-17, PI3K-AKT, TGF-β, and Hippo signaling pathways were specifically expressed in different cells after compression.