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Year : 2019  |  Volume : 14  |  Issue : 12  |  Page : 2104-2111

Novel circular RNAs expressed in brain microvascular endothelial cells after oxygen-glucose deprivation/recovery

1 Department of Anesthesiology, Huashan Hospital, Fudan University, Shanghai, China
2 Department of Medical Ultrasound, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
3 Institute of Life Sciences, Chongqing Medical University; Chongqing Foreign Language School, Chongqing, China
4 Department of Cerebrovascular Diseases, The Second Affiliated Hospital of Kunming Medical University, Kunming, Yunnan Province, China
5 Institute of Life Sciences, Chongqing Medical University, Chongqing, China

Correspondence Address:
Fei-Fei Shang
Institute of Life Sciences, Chongqing Medical University, Chongqing
Ying-Wei Wang
Department of Anesthesiology, Huashan Hospital, Fudan University, Shanghai
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Source of Support: This work was supported by the National Natural Science Foundation for Young Scientists of China, No. 81601058 (to WL); and Basic Research and Frontier Science Exploration Foundation of Yuzhong District, Chongqing, China, No. 20180106 (to FFS), Conflict of Interest: None

DOI: 10.4103/1673-5374.262589

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Circular RNAs (circRNAs) are generated by head-to-tail splicing and are ubiquitously expressed in all multicellular organisms. Their important biological functions are increasingly recognized. Cerebral ischemia reperfusion injury-induced brain microvascular endothelial cell dysfunction is an initial stage of blood-brain barrier disruption. The expression profile and potential function of circRNAs in brain microvascular endothelial cells is unknown. Rat brain microvascular endothelial cells were extracted and cultured in glucose-free medium for 4 hours with 5% CO2 and 95% N2, and the medium was then replaced with complete growth medium for 6 hours. The RNA in these cells was then extracted. The circRNA was identified by Find_circ and CIRI2 software. Functional and pathway enrichment analysis of genes that were common to differentially expressed mRNAs and circRNA host genes was performed by the Database for Annotation, Visualization and Integrated Discovery Functional Annotation Tool. Miranda software was used to predict microRNAs that were potentially spong-ed by circRNAs. Furthermore, cytoscape depicted the circR-NA-microRNA interaction network. The results showed that there were 1288 circRNAs in normal and oxygen-glucose deprived/recovered primary brain microvascular endothelial cells. There are 211 upregulated and 326 downregulated differentially expressed circRNAs. The host genes of these differentially expressed circRNAs overlapped with those of differentially expressed mRNAs. The shared genes were further studied by functional enrichment analyses, which revealed that circRNAs may contribute to calcium ion function and the cyclic guanosine 3′,5′-monophosphate (CAMP) dependent protein kinase (PKα) signaling pathway. Next, quantitative reverse transcription polymerase chain reaction assays were performed to detect circRNA levels transcribed from the overlapping host genes. Eight out of the ten circRNAs with the highest fold-change identified by sequencing were successfully verified. Subsequently, the circRNA-microRNA interaction networks of these eight circRNAs were explored by bioinformatic analysis. These results demonstrate that altered circRNAs may be important in the pathogenesis of cerebral ischemia reperfusion injury and consequently may also be potential therapeutic targets for cerebral ischemia diseases. All animal experiments were approved by the Chongqing Medical University Committee on Animal Research, China (approval No. CQMU20180086) on March 22, 2018.

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