MiR-21b-3p protects NS2OY cells against oxygen-glucose deprivation/reperfusion-induced injury by down-regulating cyclooxygenase-2

American Journal of Translational Research

Huai Huang¹  Xiaona Wu²  Kairun Peng³  Zhensheng Li⁴ 

[1] Department Two of Nerve Rehabilitation, Guangzhou General Hospital, Guangzhou Military Region, Guangzhou, Guangdong, Peoples Republic of China;Department of Neurology, General Hospital of Southern Theater Command, PLA, Guangzhou, Guangdong, Peoples Republic of China;Department of Orthopaedics, General Hospital of Southern Theater Command, PLA, Guangzhou, Guangdong, Peoples Republic of China;General Hospital of Northern Theater Command, PLA, Shenyang, Liaoning, Peoples Republic of China

关键词: Oxygen/glucose deprivation and reperfusion;    cyclooxygenase-2;    miR-21b-3p;    cell viability;    apoptosis;   

DOI  :  

学科分类：医学（综合）

来源: e-Century Publishing Corporation

【 摘 要 】

Recent studies have shown abnormal expression levels of cyclooxygenase-2 (COX-2) and miR-21b-3p in cerebral ischemia-reperfusion (I/R) rat models. Decreased COX-2 expression could reduce brain injury and thus could be a target of miR-21b-3p according to the miRNA databases (miRDB) analysis. However, its functions and underlying mechanisms in I/R injury remain unclear. In our study, we have established an oxygen/glucose deprivation and reperfusion (OGD/R) model by using NS2OY cells. The expression of miR-21b-3p and COX-2 was determined by quantitative real-time PCR or Western blot, and the fluorescence intensities were detected by fluorescence in situ hybridization (FISH) or immunofluorescence. After transfection and OGD/R treatments, the functions of miR-21b-3p and COX-2 on cell viability and apoptosis were detected using cell-counting kit 8, Edu staining, flow cytometry and Hoechst staining, respectively. Finally, dual-luciferase reporter assay was used to explore the relationship between miR-21-b-3p and COX-2. The results have showed that COX-2 mRNA and protein expression were significantly increased; however, the expression of miR-21b-3p was remarkably reduced in NS2OY cells after OGD/R treatment. The changes were most remarkable in OGD 2 h/R24 group. Function analysis has showed that when NS2OY cells were exposed to OGD/R injury, overexpressed miR-21b-3p significantly downregulated COX-2 expression, increased cell viability and decreased apoptosis. In addition, knocking down the expression of COX-2 could also increase cell viability and decrease apoptosis. Dual-luciferase reporter assays showed miR-21b-3p as the target of 3’-UTR of COX-2. Therefore, we concluded that OGD/R-induced injury by down-regulating COX-2.

【 授权许可】

CC BY-NC