Neonatal hypoxic-ischemia encephalopathy (HIE) refers to hypoxic-ischemic brain damage caused by perinatal asphyxia. Increasing evidence has revealed the crucial roles of microRNAs (miRNAs) in neonatal HIE. In the current research, we aimed to explore the biological role of miR-363-3p in neonatal HIE. For this purpose, we established in vitro models of PC-12 and SH-SY5Y cells subjected to oxygen-glucose deprivation and reperfusion (OGD/R) and an in vivo rat model subjected to middle cerebral artery occlusion/reperfusion (MCAO/R) treatment. First, using H&E staining, TTC staining, and western blot analysis, we observed that DUSP5 knockdown suppressed HIE in vivo. Then, by performing flow cytometric analysis, western blotting, RT-qPCR, and MTT assays, we observed that DUSP5 silencing suppressed OGD/R-induced cell injury in vitro. Subsequently, we explored the potential regulatory mechanism of DUSP5 in OGD/R-treated cells with luciferase reporter assays and RT-qPCR analysis. The results demonstrated that DUSP5 was targeted by miR-363-3p. Next, functional assays, including flow cytometric analysis, MTT assays, western blotting and RT-qPCR, were conducted to explore the biological functions of miR-363-3p in SH-SY5Y and PC-12 cells. Our data showed that miR-363-3p overexpression suppressed OGD/R-induced cell injury. Finally, the results from rescue experiments showed that enhanced DUSP5 expression counteracted the effect of miR-363-3p overexpression. In conclusion, our data suggested that miR-363-3p attenuates neonatal HIE by targeting DUSP5.

新生儿缺氧缺血性脑病（HIE）是指由围产期窒息引起的缺氧缺血性脑损伤。越来越多的证据揭示了 microRNA (miRNA) 在新生儿 HIE 中的关键作用。在目前的研究中，我们旨在探索 miR-363-3p 在新生儿 HIE 中的生物学作用。为此，我们建立了 PC-12 和 SH-SY5Y 细胞进行氧-葡萄糖剥夺和再灌注 (OGD/R) 的体外模型和大脑中动脉闭塞/再灌注 (MCAO/R)的体内大鼠模型。治疗。首先，使用 H&E 染色、TTC 染色和蛋白质印迹分析，我们观察到 DUSP5 敲低在体内抑制 HIE. 然后，通过流式细胞仪分析、蛋白质印迹、RT-qPCR 和 MTT 分析，我们观察到 DUSP5 沉默在体外抑制了 OGD/R 诱导的细胞损伤. 随后，我们通过荧光素酶报告基因分析和 RT-qPCR 分析探索了 DUSP5 在 OGD/R 处理的细胞中的潜在调控机制。结果表明DUSP5被miR-363-3p靶向。接下来，进行功能分析，包括流式细胞术分析、MTT 分析、蛋白质印迹和 RT-qPCR，以探索 miR-363-3p 在 SH-SY5Y 和 PC-12 细胞中的生物学功能。我们的数据显示 miR-363-3p 过表达抑制了 OGD/R 诱导的细胞损伤。最后，拯救实验的结果表明，增强的 DUSP5 表达抵消了 miR-363-3p 过表达的影响。总之，我们的数据表明 miR-363-3p 通过靶向 DUSP5 减弱新生儿 HIE。