Department of Rehabilitation Medicine, Dapeng New District Nan'ao People's Hospital, Shenzhen, 518121, People's Republic of China.
Physical Medicine and Physiotherapy, The Second School of Clinical Medicine, Southern Medical University, Guangzhou, 510515, People's Republic of China.
Metab Brain Dis. 2021 Dec;36(8):2495-2504. doi: 10.1007/s11011-021-00744-1. Epub 2021 Oct 11.
Non-coding RNAs have recently attracted much attention with the potential in the treatment of cerebral ischemia/reperfusion (I/R) injuries. In this study, we investigated the role of miR-32-5p in cerebral I/R injuries by using oxygen-glucose deprivation/reperfusion (OGD/R) PC12 cells and middle cerebral artery occlusion/reperfusion (MCAO/R) rats. The expression of genes and proteins were detected by RT-qPCR and Western blot, respectively. The function of OGD/R PC12 cells was detected using MTT assay and flow cytometry analysis. The influences of MCAO/R on rats was evaluated by measuring the infarct volume and brain water content. Bioinformatics analysis and luciferase gene reporter assay were used to identify the relationship between miR-32-5p and PTEN. The results showed that miR-32-5p had neuroprotective effects on OGD/R induced PC12 cells and MCAO/R injured rats' brain. The level of miR-32-5p was significantly reduced after OGD/R. Overexpression of miR-32-5p significantly reduced MCAO/R-induced brain damages in rats. Moreover, PTEN was found to be a target of miR-32-5p, and overexpression of PTEN attenuated the effects of miR-32-5p overexpression on cerebral I/R injuries. In addition, miR-32-5p was able to activate PI3K/AKT signaling by inhibiting PTEN. In conclusion, miR-32-5p prevents brain I/R injuries through modulating PTEN/PI3K/AKT signaling pathway.
非编码 RNA 最近受到了广泛关注,它们具有治疗脑缺血/再灌注(I/R)损伤的潜力。在这项研究中,我们通过氧葡萄糖剥夺/再灌注(OGD/R)PC12 细胞和大脑中动脉闭塞/再灌注(MCAO/R)大鼠模型来研究 miR-32-5p 在脑 I/R 损伤中的作用。分别通过 RT-qPCR 和 Western blot 检测基因和蛋白的表达。通过 MTT 测定和流式细胞术分析检测 OGD/R PC12 细胞的功能。通过测量梗死体积和脑水含量评估 MCAO/R 对大鼠的影响。通过生物信息学分析和荧光素酶基因报告基因实验来鉴定 miR-32-5p 和 PTEN 之间的关系。结果表明,miR-32-5p 对 OGD/R 诱导的 PC12 细胞和 MCAO/R 损伤大鼠的脑具有神经保护作用。OGD/R 后 miR-32-5p 的水平显著降低。过表达 miR-32-5p 可显著减轻 MCAO/R 诱导的大鼠脑损伤。此外,发现 PTEN 是 miR-32-5p 的靶基因,过表达 PTEN 可减弱 miR-32-5p 过表达对脑 I/R 损伤的作用。此外,miR-32-5p 通过抑制 PTEN 激活 PI3K/AKT 信号通路。总之,miR-32-5p 通过调节 PTEN/PI3K/AKT 信号通路来预防脑 I/R 损伤。