NOD-like-receptor signaling pathway mediates inflammation and triggers cerebral injury in cerebral venous thrombosis

Abstract

Background Cerebral venous thrombosis (CVT) is a special type of stroke with an increasing incidence. However, the pathophysiological mechanisms remain elusive, which hinders a comprehensive understanding of CVT. We used a CVT model in rats to elucidate the mechanism of neurological damage. Methods We constructed a CVT model to examine neurological function and performed neuroimaging. RNA-Seq and biological information technology were utilized to analyze the transcriptome features of the Sham, middle cerebral artery occlusion (MCAO), and CVT groups, subsequently selecting significantly upregulated signaling pathways in the CVT rat brain. The activation of signaling pathways and immune cells in CVT was confirmed through flow cytometry (FC), real-time quantitative polymerase chain reaction (qPCR), and immunofluorescence staining (IF). Results Twenty-four hours after CVT establishment, rats exhibited significant Magnetic resonance imaging (MRI)-T2 hyperintensity and neurological impairment compared to sham rats. Transcriptome profiling showed that the inflammatory response was a significant and specific characteristic of the CVT group compared with the sham and MCAO groups. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of differentially expressed genes (DEGs) indicated that the DEGs were mainly enriched in the gene set of inflammation-related responses. Single-sample gene set enrichment analysis (ssGSEA) also suggested that the immune inflammatory response score was increased significantly. Furthermore, Immune-AI mouse revealed that microglia were the most significantly elevated immune inflammatory cells after CVT. GSEA indicated that the nucleotide-binding oligomerization domain (NOD)-like-receptor signaling pathway was significantly upregulated compared to other inflammatory signaling pathways, and then, key driver analysis (KDA) of DEGs in the NOD-like-receptor signaling pathway revealed that Nod-2 and other genes were the core genes. Importantly, inhibiting the NOD-like-receptor signaling pathway in CVT rats resulted in neurological function improvement and infarct size reduction. Conclusions The microglia-mediated inflammatory response and NOD-like-receptor signaling pathway activation are significant pathological changes in brain injury after CVT. This study may enhance comprehension of the pathological mechanisms underlying CVT and provide novel insights for further investigation into injuries in CVT.