Exploration of the mechanisms affecting ischemic stroke by ceRNA network construction

Abstract

Abstract Background Ischemic stroke is among the most common and fatal diseases.. Apoptosis exerts a crucial role in ischemic stroke and heart disorders. The role of the circRNA associated with apoptosis in ischemic stroke was not fully elucidated. Methods Three datasets, GSE122709, GSE133768, and GSE110993, from the GEO database, were enrolled in bioinformatics analysis. The targeting miRNAs and mRNAs were predicted through the circBase and miRWalk database. The circRNA-miRNA-mRNA network was created by Cytoscape software. The ‘clusterProfiler’ R package was applied to execute GO and KEGG analysis. We constructed the SH-SY5Y oxygen-glucose deprivation (OGD) model to simulate ischemic stroke in vitro. RT-qPCR and western-blot was utilized to examine the expression level of corresponding genes. Cell Counting Kit-8 was utilized to assess cell viability. Flow cytometry was conducted for cell apoptosis analysis. Results Based on the data from public database, we finally created a circRNA-miRNA-mRNA network containing 143 nodes and 272 edges. In the network, the expression of CDKN1A (cyclin dependent kinase inhibitor 1A) was regulated by hsa-miR-17-5p, which is regulated by hsa-circ-0004622. Then, we verified the expression of CDKN1A, miR-17-5p, and circ-0004622 in SH-SY5Y OGD model. The result of functional experiment showed that circ-0004622 promoted apoptosis in ischemic stroke by regulating miR-17-5p. Conclusion Taken together, by bioinformatics methods and functional experiments, this study excavated and investigated the role of circ-0004622 in promoting cell apoptosis in ischemic stroke, providing a theoretical foundation for clinical diagnosis and elucidation of the molecular mechanism of ischemic stroke.