Journal of visualized experiments : JoVE · 2025
Analyses of the Competing Endogenous RNA Network in Rats with Middle Cerebral Artery Occlusion Based on lncRNA, miRNA, and mRNA Expression.
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Li Shuangping, Ye Qingping, Li Yumeng, Li Jingjing, Peng Daiyin, Duan Xianchun, Hong Shizhong
Аннотация
This study aims to investigate the regulatory roles and underlying mechanisms of lncRNAs acting as ceRNAs in ischemic stroke. Based on the ceRNA hypothesis, lncRNAs, miRNAs, and mRNAs were identified as components of a regulatory network involved in stroke. Key lncRNAs from the resulting subnetwork were selected for detailed analysis. Functional enrichment analysis using Gene Ontology and pathway mapping through the Kyoto Encyclopedia of Genes and Genomes revealed critical interactions within the lncRNA-associated ceRNA network. Key pathways, including calcium signaling, gap junction signaling, and neuroactive ligand receptor interaction, were further validated using Western blot analysis. The constructed ceRNA network comprised 334 lncRNAs, miRNAs, and mRNAs, with functional enrichment analysis predicting their biological roles. Three lncRNAs with high degree centrality were selected to construct a representative ceRNA subnetwork. Western blot analysis revealed that, compared to the sham group, the expression levels of key proteins -- CaMKII, calmodulin, CX36, PKC, CX43, GRIA3, GABRA6, and NPY1R were significantly downregulated in the model group, while the expression of CaN was significantly upregulated (P < 0.05). These findings suggest that lncRNAs are significantly involved in stroke pathogenesis. In conclusion, lncRNAs acting as ceRNAs serve critical regulatory roles in the pathogenesis of ischemic stroke.